Liver transcriptome response to periparturient hormonal and metabolic changes depends on the postpartum occurrence of subacute ruminal acidosis in Holstein cows.

We investigated changes in rumen fermentation, peripheral blood metabolites and hormones, and hepatic transcriptomic dynamics in Holstein cows with and those without subacute ruminal acidosis (SARA) during the periparturient period. Sixteen multiparous Holstein cows were categorized in the SARA (n = 8) or non-SARA (n = 8) groups depending on whether they developed SARA during the 2 wk after parturition. Reticulo-ruminal pH was measured continuously throughout the study. Rumen fluid, blood, and liver tissue samples were collected at 3 wk prepartum and 2 and 6 wk postpartum, with an additional blood sample collected at 0 and 4 wk postpartum. The 1-h mean pH was depressed postpartum in both groups, whereas depression was more severe in the SARA group simultaneously with significantly longer duration of time (for pH <5.6 and 5.8). Significant expression of differentially expressed genes in liver tissue (DEGs; false discovery rate corrected P < 0.1) were identified only in the non-SARA group and were further analyzed by Ingenuity Pathway Analysis software. Among the top expressed DEGs, the hepatic genes encoding lipid and cholesterol secretion (APOA1, APOA4, and G0S2) and gluconeogenesis (PC, G6PC, and PCK1) were upregulated postpartum. In silico analysis revealed the significant postpartum activation of upstream regulators, such as INSR, PPARG, and PPARGC1A. These results suggested that hepatic transcriptomic responsiveness to postpartum metabolic load and hormones were likely discouraged in cows with SARA when compared with the significant activation of genes and signaling pathways for adequate metabolic adaption to postpartum high-grain diet feeding in Holstein cows without SARA.

Notice of RETRACTION: Changes in rumen fermentation, bacterial community, and predicted functional pathway in Holstein cows with and without subacute ruminal acidosis during the periparturient period.

The authors have elected to retract this paper in accordance with the following points. The article as written contains misleading information and omits important details. Cows in this study were assigned to groups based on the current definition of subacute ruminal acidosis; they were housed on two different farms and fed two different sets of rations in this study. However, multiple farms were not described in the materials and methods and this was not accounted for in the statistical analysis as published. The diets shown in Table 1 were not actually fed to animals; rather, the proportions of ingredients listed represent an average of the two farms housing the cows. The authors regret the errors.

Effects of pulse-dose ruminal infusion of butyrate on plasma glucagon-like peptide 1 and 2 concentrations in dairy calves.

Feeding of butyrate was found to have a positive effects in enhancing gut development and improving growth performance of calves. Equally, glucagon-like peptide 1 and 2 (GLP-1 and GLP-2), secreted from gastrointestinal L-cells in response to nutrient intake, were found to play a significant role in regulating blood glucose homeostasis and improving gut health. However, limited information is available about the relationship between butyrate and release of GLP-1 and GLP-2 in dairy calves. The objective of this study was to evaluate the effects of a pulse-dose ruminal infusion of butyrate on plasma GLP-1 and GLP-2 concentrations in dairy calves. Five ruminally cannulated mature Holstein bull calves (7.2 ± 0.10 mo, and 330 ± 16.0 kg of body weight; mean ± standard deviation) were used in a 5 × 5 Latin square with 4-d periods. On d 1 of each period at 0800 h, calves were ruminally infused with 1 of 5 treatments: 0 (saline), 0.3, 0.6, 0.9, and 1.2 g of butyrate per kg of body weight. Before butyrate infusion, calves were not offered feed overnight, and sequential blood and rumen fluid samples were taken before and after infusion on d 1 of each period. Ruminal butyrate and total volatile fatty acid concentrations increased linearly (2.65, 12.19, 20.99, 30.19, and 36.30; 23.68, 33.07, 40.94, 51.13, and 56.31 µmol/mL, for butyrate and total volatile fatty acids, respectively) in a dose-dependent manner, whereas propionate and isobutyrate increased quadratically. Ruminal and plasma butyrate, ß-hydroxybutyrate, GLP-1, GLP-2, insulin, and glucose concentrations were all affected by treatment, time (except GLP-2), and interaction of treatment with time (except GLP-1). The area under the curve (AUC) summarized at different time points relative to the baseline (AUC30, AUC60, AUC120, and AUC240) for ruminal and plasma butyrate, and BHB, increased linearly with the dose of butyrate infused. However, AUC30, AUC60, AUC120, and AUC240 for plasma GLP-2 concentration were affected in a cubic manner unlike the linear effect on AUC30 and AUC60 for GLP-1. Plasma GLP-2 was not correlated with plasma butyrate (r = 0.16), GLP-1 (r = 0.03), or BHB (r = -0.05). This findings suggest that pulse-dosing of butyrate slightly increased both GLP-1 and GLP-2 concentrations at specific time points and this might be promoted by direct or indirect effect of butyrate on the intestinal L-cells.

Effect of milking time on yield, composition, and antimicrobial components of milk in lactating goats.

Various studies have attempted to improve the milk yield and composition in dairy animals. However, no study has examined the effects of milking at different times on milk yield and composition. Therefore, this study aimed to compare the yield, composition, and antimicrobial components of milk obtained from milking at different times in lactating goats. Eight goats were milked once daily at different times for three consecutive weeks (first week: 06:00 h; second week: 09:00 h; and third week: 12:00 h). The light ranged from 06:30 to 19:00 h. Milk and blood samples were collected once a day during milking time. Milking at 09:00 h resulted in a significantly higher milk yield than that obtained after milking at 06:00 and 12:00 h. Prolactin levels in plasma and the fat, Na+, ß-defensin, and S100A7 (antimicrobial component) levels in milk were the lowest in the 09:00 h milking. These results indicate that milk yield, composition, and antimicrobial components can be affected by milking time, which may be related to the altered concentration of prolactin in the blood. These findings provide a rational basis for achieving maximal milk production with strong immunity by changing to a more effective milking time.

Effect of intramammary lipopolysaccharide challenge after repeated intrauterine infusion of lipopolysaccharide on the inflammation status of goat mammary glands.

Previous studies have shown that a single infusion of lipopolysaccharide (LPS) into the uterus induces mammary gland inflammation. However, repeated LPS infusions return the mammary glands to their basal state of inflammation. To confirm that this is a state of tolerance to LPS, we examined whether tolerance induced by repeated intrauterine LPS infusions limits mammary gland inflammation following subsequent intramammary LPS infusions. In the first experiment, three goats were treated with repeated intrauterine infusions of LPS dissolved in black ink for 5 consecutive days. Blood and milk samples were collected at 2, 4, 6, 12, 24, 48, 72, 96, and 120 h and smeared on glass slides to confirm the translocation of LPS from the uterus to the mammary gland. Black particles were detected in the blood and milk samples more than 2 h after the first infusion and in the connective tissue of the mammary gland after day 5. In the second experiment, goats were divided into two groups: an intrauterine infusion group (IU; n = 7) and a control group (CON; n = 6). The IU group received an intrauterine infusion of 100 µg of LPS in saline for 5 days. Subsequently, LPS was infused into the mammary glands of both groups to examine the effect of intrauterine treatment on the mammary inflammatory response after intramammary LPS infusion. Blood and milk samples were collected at 6, 12, and 24 h, and then daily until 7 d after the intramammary LPS challenge. Interestingly, a significant increase in the milk somatic cell count (SCC), IL-8, IL-1ß, and TNF-α concentrations were observed in the CON group compared to the IU group. This suggests that pretreatment with repeated intrauterine infusions of LPS suppresses the inflammatory responses to subsequent intramammary LPS challenges.

Comparison of oral administration of fructose and glucose on food intake and physiological parameters in broiler chicks.

Like glucose, fructose is a monosaccharide, but the mechanisms of its absorption and metabolism in the body are very different between the 2 molecules. In this study, we investigated the effects of oral administration of glucose and fructose on food intake, diencephalic gene expression, and plasma metabolite concentrations in broiler chicks. The animals used in this study were 4-day-old male broiler chicks (Ross 308). They were given glucose, fructose (200 mg/ 0.5 mL/ bird), or a similar volume of distilled water orally after 6 h fasting. After treatment, measurements of food intake (at 0, 30, and 60 min), and blood glucose as well as insulin concentrations were measured over time; however, diencephalic (hypothalamus) gene expression and plasma metabolites were measured at 30 min. The results showed that glucose administration suppressed food intake, but fructose administration did not suppress food intake and it was at the same level as distilled water administration. In addition, fructose administration did not increase plasma glucose and insulin levels as did glucose administration. In the diencephalon, expression levels of genes related to the melanocortin system were unaffected by the treatment, while gene expression levels related to intracellular energy regulation, such as AMP-activated protein kinase were affected by the glucose treatment in the fasted chicks. These results suggest that fructose administration does not suppress feeding behavior as a result of possible reduction in the energy levels in the diencephalon and associated energy metabolism.

Effect of Ca-octanoate supplementation on concentrations of ghrelin and ghrelin-related hormones in plasma and milk of beef cattle.

We aimed to evaluate the effect of dietary calcium (Ca)-octanoate supplementation on concentrations of ghrelin, growth hormone (GH), insulin-like growth factor-1 (IGF-1), and insulin in plasma and milk of beef cattle during late gestation and early postpartum. Twelve Japanese Black cattle were offered concentrate without (CON, n = 6) or with Ca-octanoate supplementation at 1.5% of dietary dry matter (OCT, n = 6). Blood samples were collected at -60, -30, and -7 d relative to the expected parturition date and daily from d 0 to 3 after parturition. Milk samples were collected daily postpartum. Compared to the CON group, concentrations of acylated ghrelin increased in plasma as parturition approached in the OCT group (P = 0.02). However, concentrations of GH, IGF-1, and insulin in plasma and milk were not affected by treatment groups throughout the study. Additionally, we showed for the first time that bovine colostrum and transition milk contain acylated ghrelin at a significantly higher concentration than plasma (P = 0.01). Interestingly, concentrations of acylated ghrelin in milk were negatively correlated with those in plasma postpartum (r = -0.50, P < 0.01). Feeding Ca-octanoate increased concentrations of total cholesterol (T-cho) in plasma and milk (P < 0.05), tended to increase those of glucose in plasma at postpartum and milk (P < 0.1). We conclude that feeding Ca-octanoate in late gestation and early postpartum may contribute to increased concentrations of glucose and T-cho in plasma and milk without affecting concentrations of ghrelin, GH, IGF-1, and insulin in plasma and milk.

Effect of ensiled total mixed rations containing sake lees on digestion, nitrogen use, and plasma metabolite concentrations in sheep.

Sake lees are a byproduct of Japanese rice wine and used as ruminant feed with high protein and ethanol contents. This study aimed to explore the effects of ensiled total mixed ration (TMR) containing sake lees on rumen fermentation, digestion, nitrogen (N) use, and plasma metabolites in sheep. Four mature wethers were assigned to a 4 × 4 Latin square design with a factorial arrangement of ensiling treatment (non-ensiled or ensiled TMR) and types of sake lees (traditional or liquefied). Although dietary ethanol consumption was higher in sheep fed ensiled TMR, ruminal and plasma concentrations of methanol and ethanol increased after feeding in all dietary treatments, which was accompanied by the increased plasma concentrations of formate. The intake and digestion of dry matter and N were lower in sheep fed ensiled TMR than in those fed non-ensiled TMR. The plasma concentrations of most amino acids decreased in sheep fed ensiled TMR. Although plasma methionine, serine, and glycine concentrations were not altered by diet, homocysteine concentration was the highest in sheep fed ensiled TMR containing traditional sake lees. The negative impacts of ensiling treatment on N digestion and amino acid utilization should be considered for formulating ensiled TMR containing sake lees.

Parturition and postpartum dietary change altered ruminal pH and the predicted functions of rumen bacterial communities but did not alter the bacterial composition in Holstein cows.

We investigated the temporal dynamics of ruminal pH and the composition and predicted functions of the rumen bacterial community in Holstein cows during the periparturient period. Eight multiparous Holstein cows (body weight; 707.4 ± 29.9 kg, parity; 3.6 ± 0.6) were used for continuous reticulo-ruminal pH measurement, among which five were also used for bacterial community analysis. Rumen fluid samples were collected at 3 weeks before and 2 and 6 weeks after parturition, and blood samples were collected 3 weeks before and 0, 2, 4, and 6 weeks after parturition. After the parturition, reduction in the 1-h mean reticulo-ruminal pH was associated with a significant (P < 0.05) increase in total volatile fatty acid concentration. However, with the exception of a significant change in an unclassified genus of Prevotellaceae (P < 0.05), we detected no significant changes in the compositions of major bacterial phyla or genera or diversity indices during the periparturient period. On the basis of predicted functional analysis, we identified a total of 53 MetaCyc pathways (45 upregulated), 200 enzyme commissions (184 upregulated), and 714 Kyoto Encyclopedia of Genes and Genomes orthologs (667 upregulated) at 6 weeks postpartum that were significantly (P < 0.05) different to those at 3 weeks prepartum. Among the 14 MetaCyc pathways (P < 0.05) in which pyruvate is an end product, PWY-3661 [log2-fold change (FC) = 5.49, false discovery rate (FDR) corrected P < 0.001] was the most highly upregulated pyruvate-producing pathway. In addition, P381-PWY [adenosylcobalamin biosynthesis II (aerobic); FC = 5.48, FDR corrected P < 0.001] was the second most upregulated adenosylcobalamin (Vitamin B12)-producing pathway. In contrast, PWY-2221 (FC = -4.54, FDR corrected P = 0.003), predominantly found in pectinolytic bacteria, was the most downregulated pathway. Our findings indicate that changes in rumen bacterial community structure are not strictly associated with transitions in parturition or diet; however, we did observe changes in reticulo-ruminal pH and the metabolic adaptation of predicted functional pathways. Consequently, predictive analysis of postpartum functional pathways may enable us to gain insights into the underlying functional adaptations of bacterial communities in Holstein cows during the periparturient period.

Anaerobic fermentation of rice bran using rumen liquor for desirable chemical changes as animal feed.

Objective: The objectives of this research are to overcome the limitations of rice bran (RB) and de-oiled rice bran (DORB) by fermentation anaerobically using inoculum from the rumen of a canulated sheep for desirable chemical changes. Materials and Methods: Initially, RB and DORB were fermented by 10% rumen liquor for 12 h at 39°C at different moisture levels (10, 20, 30, 40, 50, and 60% phosphate buffer). Again, DORB was fermented for 24, 48, and 72 h at 39°C using 10% rumen liquor at different moisture levels (10, 20, 30, 40, 50, and 60% phosphate buffer). Before and after fermentation, RB and DORB were analyzed for pH, proximate components, neutral detergent fiber (NDF), total-P, inorganic-P, and phytate-P. Results: Fermentation of RB and DORB for 12 h reduced (p < 0.05) pH, crude fiber (CF),NDF, and phytate-P, but increased (p < 0.05) the content of inorganic-P. Subsequent fermentation of DORB for 24, 48, and 72 h reduced pH, CF, and NDF. Total-P of fermented DORB remained similar till 72 h fermentation (p < 0.05). But, inorganic-P increased with the increasing duration (24, 48, and 72 h) of fermentation and increased (30, 40, and 50) moisture level (p < 0.05). Alternatively, phytate-P decreased with increasing duration and moisture level (p < 0.05). Conclusion: Inoculation of rumen microbes and incubation of RB (12 h) and DORB (24 h) at room temperature reduced phytate-P and fiber content (CF and NDF) when the moisture level was up to 50%; those are the indicators to reduce the limitation of RB and DORB to use as feed for non-ruminant animals like poultry and pigs.

Effects of wood kraft pulp as a partial replacement for roughage on rumen fermentation and productivity in dairy cows.

In this study, we evaluated the partial replacement of roughage with wood kraft pulp (KP) on rumen fermentation and productivity of dairy cows. Eighteen cows were divided into control and KP groups. The KP group started adaptation to KP 3 weeks before calving; after calving, they were fed a total mixed ration for 12 weeks, wherein 18% Timothy hay was replaced with KP. The dry matter intake, body weight, and milk yield and composition were similar in the control and KP groups. The average daily rumen pH was higher with KP feeding, and the average daily ruminal temperature remained lower at 16 days after calving (P < 0.05). The concentration of volatile fatty acids remained unaltered, the molar proportion of acetic acid decreased, and the molar proportion of propionic acid increased, indicating a low acetic acid:propionic acid ratio (P < 0.05). Lipopolysaccharide activity in the rumen fluid was higher in the KP group (P < 0.05); however, the rumen microbiota were unaffected. The digestibility of dry matter and neutral detergent fiber increased 12 weeks after calving, whereas excretion of urinary nitrogen decreased (P < 0.05). Partial replacement of roughage with KP did not suppress rumen fermentation and maintained postpartum productivity.

Follicular persistence induced by adrenocorticotropic hormone administration in goats.

The objective of the present study was to examine the effect of adrenocorticotropic hormone (ACTH) administration on the induction of persistent cystic follicle in the goat in order to establish a method to experimentally induce cystic follicle. Four cross-bred goats were intramuscularly administered ACTH at 0.78 and 6.25 µg/10 kg twice a day from Days 15 to 21 (Day 0 was defined as the day of last estrus). Follicular status in the ovary was monitored by ultrasound examination. The plasma concentrations of estradiol, progesterone and cortisol were measured. Treatment with ACTH at the 0.78 and 6.25 µg/10 kg levels caused persistent follicles (> 10 days delay from the expected ovulation date) in 50% of the goats in both treatment groups. In those animals, ovulation occurred 17 and 27 days and 11 and 12 days after the expected days in the 0.78 and 6.25 µg/10 kg groups, respectively. The maximum follicle diameters were 10 and 9 mm in the 0.78 and 6.25 µg/10 kg ACTH groups, respectively. In the control group, the estradiol concentration increased on Day 18 and remained at a high level for a few days. However, such an increase was not seen in both ACTH groups. The estradiol concentration increased gradually from Days 21 to 27 in the 6.25 µg/10 kg ACTH group. These results suggest the possibility that ACTH induces persistent follicles in goats, which may be related to the delay of the onset of estradiol secretion followed by its maintenance at a high concentration.

Effect of ensiling persimmon peel and grape pomace as tannin-rich byproduct feeds on their chemical composition and in vitro rumen fermentation.

This study aimed to evaluate the effects of ensiling fruit byproducts on their chemical composition and in vitro ruminal fermentation. Persimmon peel (PP), white grape pomace (WGP), and red grape pomace (RGP) were ensiled for 1, 2, 4, and 8 weeks. Fresh and ensiled PP, WGP, and RGP were used for in vitro rumen fermentation with or without polyethylene glycol (PEG). The non-fiber carbohydrate (NFC) content of the byproducts decreased after ensiling, especially for PP. The total tannin content was not changed after ensiling for up to 4 weeks for all byproducts. However, the soluble tannin content in PP decreased but that in grape pomaces was unaffected by ensiling. Gas production, total volatile fatty acid concentration, and methane production by in vitro rumen fermentation for PP and WGP were reduced by ensiling, and increased by adding PEG, except for gas production from the PP silage. These results indicated that changes in the fractions of carbohydrate and tannins during the ensiling process were different between PP and grape pomace. Even though the insolubilization of tannins in PP during ensiling reduced its inhibitory effect on ruminal fermentability, the ensiling PP seemed to remain the ability to mitigate methanogenesis in the rumen.

Effect of phytol in forage on phytanic acid content in cow's milk.

OBJECTIVE: Bioactive compounds in ruminant products are related to functional compounds in their diets. Therefore, this study aimed to explore the effect of forage sources, Italian ryegrass (IR) silage vs corn silage (CS) in the total mixed ration (TMR), on milk production, milk composition, and phytanic acid content in milk, as well as on the extent of conversion of dietary phytol to milk phytanic acid. METHODS: Phytanic acid content in milk was investigated for cows fed a TMR containing either IR silage or CS using 17 cows over three periods of 21 days each. In periods 1 and 3, cows were fed CS-based TMR (30% CS), while in period 2, cows were fed IR silage-based TMR (20% IR silage and10% CS). RESULTS: The results showed that there were no differences in fat, protein, lactose, solidsnot-fat, somatic cell count, and fatty acid composition of milk among the three experimental periods. There were no differences in the plasma concentration of glucose, triglycerides, total cholesterol, and nonesterified fatty acids among the three experimental periods, while the blood urea nitrogen was higher (p<0.05) in period 2. The milk phytanic acid content was higher (p<0.05) in period 2 (13.9 mg/kg) compared with periods 1 (9.30 mg/kg) and 3 (8.80 mg/kg). Also, the phytanic acid content in the feces was higher (p<0.05) in period 2 (1.65 mg/kg dry matter [DM]) compared with period 1 (1.15 mg/kg DM), and 3 (1.17 mg/kg DM). Although the phytol contents in feces did not differ among the three feeding periods, the conversion ratio from dietary phytol to milk phytanic acid was estimated to be only 2.6%. CONCLUSION: Phytanic acid content in cow's milk increases with increasing phytol content in diets. However, phytol might not be completely metabolized in the rumen and phytanic acid, in turn, might not be completely recovered into cow's milk. The change of phytanic acid content in milk may be positively correlated with the change of phytol in the diet within a short time.

Oral administration of L-citrulline changes the concentrations of plasma hormones and biochemical profile in heat-exposed broilers.

We examined the effects of oral administration of L-citrulline (L-Cit) on plasma metabolic hormones and biochemical profile in broilers. Food intake, water intake, and body temperature were also analyzed. After dual oral administration (20 mmol/head/administration) of L-Cit, broilers were exposed to a high ambient temperature (HT; 30 ± 1°C) chamber for 120 min. Oral administration of L-Cit reduced (p < .001) rectal temperature in broilers. Food intake was increased (p < .05) by heat stress, but it was reduced (p < .05) by L-Cit. Plasma levels of 3,5,3'-triiodothyronine, which initially increased (p < .0001) due to heat stress, were reduced (p < .01) by oral administration of L-Cit. Plasma insulin levels were increased by heat exposure (p < .01) and oral L-Cit (p < .05). Heat stress caused a decline (p < .05) in plasma thyroxine. Plasma lactic acid (p < .05) and non-esterified fatty acids (p < .01) were increased in L-Cit-treated heat-exposed broilers. In conclusion, our results suggest that oral L-Cit can modulate plasma concentrations of major metabolic hormones and reduces food intake in broilers.

Ensiling of total mixed ration containing persimmon peel: Evaluation of chemical composition and in vitro rumen fermentation profiles.

The effects of inclusion of persimmon peel (PP) in total mixed ration (TMR) silage on its nutrient composition, tannin content, and in vitro ruminal fermentation were studied. Four types of TMR silages containing 0, 50, 100, and 150 g/kg of PP on a dry matter basis were prepared. The dietary contents of non-fiber carbohydrate (NFC) decreased, while soluble protein fraction increased after ensiling of the TMR. In the TMR silages, the content of insoluble tannin increased (p < .05) with increasing PP level. The fraction of soluble protein decreased linearly (p < .01), while that of neutral detergent insoluble protein increased linearly (p < .01) with increasing the PP level in the TMR silages. The total gas and methane yields from the in vitro rumen fermentation of the TMR silages were lower (p < .01) than those of pre-ensiled TMR and declined linearly (p < .01) with increasing PP level. These results indicate that adding PP to TMR silage may resist the breakdown of dietary protein during the ensiling process, although the ruminal fermentability of TMR possibly decreased after ensiling due to the loss of NFC.

Effects of oral administration of colostrum whey in peripartum goat on antimicrobial peptides in postpartum milk.

The present study was conducted to examine whether colostrum supplementation in peripartum goats increases the antimicrobial peptides in their milk. Goats were orally administered 2 ml of colostrum whey products (colostrum group) or water (control group) daily, from 2 weeks before until 2 weeks after kidding. Body weights of mothers and kids were measured. Blood, milk, and fecal samples were collected from the mothers, and blood samples were collected from the kids. Concentrations of milk antimicrobial peptides (beta-defensin, cathelicidin, lactoferrin, S100A7, lactoperoxidase, and immunoglobulin A [IgA]) were determined. IgA and nutritional parameters (glucose, total cholesterol, triglyceride, ketone bodies, and non-esterified fatty acids) were also determined in the blood of mothers and kids. Milk IgA and lactoferrin concentrations were higher in the colostrum group than in the control group. Conversely, lower milk concentrations of S100A7 were observed in the colostrum group than that in the control group. Plasma IgA concentrations were higher for kids from the colostrum group than for those from the control group. These results suggest that oral administration of colostrum in pregnant goats increases IgA concentration in postpartum milk, which can subsequently improve the health of their kids.

Effect of varying fermentation conditions with ensiling period and inoculum on photosynthetic pigments and phytol content in Italian ryegrass (Lolium multiflorum Lam.) silage.

This study aimed to investigate the effect of an ensiling period (Experiment 1) and adding lactic acid bacteria (LAB, Experiment 2) on the changes in carotenoid, chlorophyll, and phytol in ensiled Italian ryegrass (IR, Lolium multiflorum Lam.). In Experiment 1, the IR herbage ensiled into plastic bags was analyzed for the contents of photosynthetic pigments and phytol over a 5-week period. During the ensiling process, the ß-carotene content decreased (p < .05), whereas the lutein content did not change. Although the chlorophyll content decreased (p < .05) after ensiling, the phytol content barely changed until week 5. In Experiment 2, IR herbage was ensiled without additive, as a Control, or with LAB for 60 days. The pH was lower (p < .05) and lactic acid content was higher (p < .05) for the LAB silage than for the Control. The chlorophyll content in silage was not affected by the LAB; however, the ß-carotene content was higher (p < .05) for the LAB silage than for the Control. Phytol and lutein contents in the herbage did not change after ensiling. These results indicate that phytol and lutein in IR herbage can be preserved well in silage, irrespective of their fermentation condition.

Changes in peripheral blood oxidative stress markers and hepatic gene expression related to oxidative stress in Holstein cows with and without subacute ruminal acidosis during the periparturient period.

We investigated changes in peripheral blood metabolites, oxidative stress markers (malondialdehyde, potential antioxidant capacity, and glutathione peroxidase [GPX]), and hepatic gene expression related to oxidative stress in Holstein cows with and without subacute ruminal acidosis (SARA) during the periparturient period. Eighteen multiparous Holstein cows were categorized into SARA (n=9) or non-SARA (n=9) groups depending on whether they developed SARA; reticulo-ruminal pH was <5.6 for more than 3 hr per day, during the 2 weeks after parturition. Blood and liver tissue samples were collected 3 weeks prepartum and 2 and 6 weeks postpartum, with an additional blood sample collected 0 and 4 weeks postpartum. Blood aspartate transaminase (AST) and nonesterified fatty acid (NEFA) increased significantly (P<0.05) after parturition in both groups. GPX activity decreased gradually after parturition in the SARA group. In the SARA group, gene expression of GPX 1 and microsomal glutathione S-transferase 3 (MGST3) decreased significantly (P<0.05), and expression of metallothionein 2A increased significantly (P<0.05) after parturition in the SARA group. Superoxide dismutase 1 and MGST3 decreased significantly (P<0.05) 2 weeks postpartum in the non-SARA group. Gene expression related to oxidative stress was negatively correlated with AST, NEFA and total ketone body levels. Therefore, the hepatic gene expression related to oxidative stress might change associated with a negative energy balance, and might relate the high oxidative stress in the SARA group during periparturient period.