Differences between pH of indwelling sensors and the pH of fluid and solid phase in the rumen of dairy cows fed varying concentrate levels

Feeding of high‐concentrate diets to cattle increases the risk of subacute ruminal acidosis (SARA). Indwelling wireless pH sensors have become popular as an early diagnostic tool for SARA recently. However, the recommended pH thresholds of SARA derive from measurements taken from free‐rumen liquid (FRL) in the ventral rumen, and not from the reticulum, where the mentioned sensors are located. The aim of this study was to evaluate commercially available indwelling pH boli for the accuracy and precision in predicting ruminal pH of FRL and the particle‐associated rumen liquid (PARL) under varying dietary concentrate levels. An additional aim was to define SARA pH thresholds of indwelled pH boli, which can be used for SARA diagnostics. The experiment was conducted with eight nonlactating rumen cannulated Holstein cows fed 0% or 65% concentrate. Data showed that the mean pH of indwelled boli was consistently higher than in FRL and PARL (pH 6.62 ± 0.02, 6.43 ± 0.02 and 6.18 ±0.02, respectively) across feeding regimens. Interestingly, the diurnal differences in pH among indwelled boli, FRL and PARL became greater during concentrate feeding, especially at 8 h after the morning feeding, suggesting that with high‐concentrate diets a particular adjustment of reticular sensor pH vs. ruminal pH in FRL and PARL is needed. The concordance correlation coefficient analysis, representing the reproducibility of the bolus measurements, was high for bolus‐FRL (0.733) and moderate for bolus‐PARL (0.510) associations. Furthermore, the quantitative relationship of the pH in FRL and PARL to the pH of the boli was described by linear regression analysis. The study determined that the common SARA threshold of pH 5.8 in FRL corresponds to a bolus pH of 6.0.     

Feeding hay rich in water‐soluble carbohydrates improves ruminal pH without affecting rumination and systemic health in early lactation dairy cows

Forages rich in water‐soluble carbohydrates (WSC) might be an ideal energy source during early lactation, as they provide both energy for milk production and structural fibre to promote chewing and rumen buffering. Thus, the aim was to investigate feeding strategies based on high‐quality hay rich in WSC with graded amounts of concentrate on ruminating behaviour, ruminal pH and systemic health variables. Twenty‐four Simmental cows were randomly allocated to four groups beginning 10 days before until 28 days after calving. Diets were 60LQH (60% fibre‐rich hay plus 40% concentrate), 60HQH, 75HQH and 100HQH (60%, 75% and 100% high‐quality hay, plus 40%, 25% and 0% concentrate, respectively). Hay qualities differed in contents of WSC (110 g vs. 198 g of dry matter [DM]), neutral detergent fibre (646 g vs. 423 g of DM) and crude protein (65 g vs. 223 g of DM). Rumination was recorded using the Rumiwatch system over 4 days during the last week. Weekly serum samples were analysed for the liver enzymes aspartate aminotransferase, glutamate dehydrogenase and γ‐glutamyltransferase and the acute phase proteins serum amyloid A and haptoglobin. Four cows per group received a wireless pH sensor orally placed into the rumen one week before the expected calving date. Daily time spent chewing did not differ between groups. Likewise differences in minimum, maximum and mean pH‐values were not significant, but daily time of reticular pH <6 was longer in cows fed 60LQH compared to cows fed 100HQH (p = 0.043) and in tendency to cows fed 75HQH or 60HQH (p = 0.072 or p = 0.086, respectively). Blood parameters were unaffected by diet. Accordingly the present results demonstrate that feeding hay rich in WSC helped stabilizing the reticuloruminal pH in early lactation dairy cows, even in combination with 40% concentrates in DM.     

Prevalence and consequence of subacute ruminal acidosis in Polish dairy herds

The aim of this study was to estimate the prevalence of subacute ruminal acidosis (SARA) in Polish high‐yielding dairy herds. Also, the relationships between the chemical composition of the diet and the feed particle size, ruminal pH and the occurrence of this metabolic disease and the fermentation profile were determined. Rumen fluid samples were obtained from a total of 213 cows from nine dairy herds (≥20 cows per herd) via rumenocentesis. Almost 14% (30/213) of cows as acidotic (pH<5.6) were found, which is indicative of SARA. Moreover, 44% of the herds were classified as SARA‐positive and 56% as SARA‐negative. Results of the current study suggested that the physically effective NDF (NDF_>1.18 mm)‐to‐starch ratio could be better indices than peNDF_>1.18 mm to preventing the occurrence of SARA, and their level should not be lower than 1.00. Also, linear negative relationships between rumen fluid pH and concentration of propionate, valerate and total VFA were determined. According to the herd's SARA status and rumen fluid biochemical indices, there were significant differences between the pH of rumen fluid (p ≤ 0.01), concentrations of acetate (p ≤ 0.05), propionate (p ≤ 0.05), n‐butyrate (p ≤ 0.01), n‐valerate (p ≤ 0.01), the sum of VFA (p ≤ 0.01) and ammonia (p ≤ 0.05) in SARA‐positive herds compared to SARA‐negative herds. The better understanding the strategy of ruminant nutrition to coordinate energy conversion and the role of the ruminal pH in regulating N‐NH₃ production, absorption through rumen mucosa, urea secretion, the more successfully we can utilize these processes with due recognition of animal needs and welfare, as well as prevention of SARA occurrence.     

Metagenomic profiles of the rumen microbiota during the transition period in low‐yield and high‐yield dairy cows

We investigated potential relationships between rumen microbiota and milk production in dairy cows during the transition period. Twelve dairy cows were divided into a low‐yield (LY) or high‐yield (HY) group based on their milk yield. Rumen samples were taken from dairy cows at 3 weeks before parturition, and at 4, 8, and 12 weeks after parturition. 16S rDNA‐based metagenomic analysis showed that diversities of rumen microbiota in both groups were similar and the number of operational taxonomic units (OTUs) was lower in the postpartum than prepartum period in both groups. The abundance of Bacteroidetes and ratio of Bacteroidetes:Firmicutes was higher in the HY than the LY group. OTUs assigned to Prevotella bryantii, Fibrobacter succinogenes, Ruminococcus albus, Butyrivibrio fibrisolvens, and Succinivibrio sp. were abundant in the HY group. These OTUs were significantly related to the propionate molar proportion of rumen fluids in the HY group. OTUs assigned to Lachnospiraceae, Bifidobacterium sp. and Saccharofermentans were dominant in the LY group. Predictive functional profiling revealed that abundance of gene families involved in amino acid and vitamin metabolism was higher in the HY than the LY group. These results suggest that the community structure and fermentation products of rumen microbiota could be associated with milk production of dairy cows.     

Effect of tannins and saponins in Samanea saman on rumen environment,milk yield and milk composition in lactating dairy cows

The objective of this study was to investigate the effects of tannins and saponins in Samanea saman on rumen fermentation, milk yield and milk composition in lactating dairy cows. Four multiparous early‐lactating dairy cows (Holstein‐Friesian cross‐bred, 75%) with an initial body weight (BW) of 405 ± 40 kg and 36 ± 8 day in milk were randomly assigned to receive dietary treatments according to a 4 × 4 Latin square design. The four dietary treatments were unsupplemented (control), supplemented with rain tree pod (S. saman) meal (RPM) at 60 g/kg, supplemented with palm oil (PO) at 20 g/kg, and supplemented with RPM at 60 g/kg and PO at 20 g/kg (RPO), of total dry matter (DM) intake. Cows were fed with concentrate diets at a ratio of concentrate to milk yield of 1:2, and chopped 30 g/kg of urea‐treated rice straw was fed ad libitum. The RPM contained condensed tannins and crude saponins at 88 and 141 g/kg of DM respectively. It was found that s upplementation with RPM and/or PO to dairy cows diets did not show negative effect on ruminal pH, blood urea nitrogen and milk urea nitrogen concentration (p > 0.05). However, supplementation with RPM resulted in lower ammonia nitrogen (NH₃‐N) concentration (p < 0.05). In addition, propionic acid and milk production increased while acetic acid, acetic to propionic ratio, methane production, methanogens and protozoal population decreased with RPM and/or PO supplementation. Furthermore, addition of PO and RPO in the diets increased milk fat while supplementation of RPM resulted in greater milk protein and Fibrobacter succinogenes numbers (p < 0.05). The population of Ruminococcus flavefaciens and Ruminococcus albus were not affected by any treatments. The findings on the present study showed that supplementation with RPM and RPO to diets of cows improved the rumen environment and increased milk yield, content of milk protein and milk fat.     

Effects of dietary Greek oregano (Origanum vulgare ssp. hirtum) supplementation on rumen fermentation,enzyme profile and microbial communities in goats

This study was conducted to examine in vivo long‐term effects of dietary dried oregano (Origanum vulgare ssp. hirtum) whole plant on rumen fermentation, enzyme profile and microbial communities. For this purpose, eight healthy, adult, non‐lactating Alpine goats were kept in tie stalls equipped for individual feeding and randomly divided into two homogeneous groups: one fed 0.6 kg of a concentrate mixture and 0.6 kg of wheat straw without any supplementation and served as control group (CON) while the other group (OR) fed the same diet of CON but supplemented with 20 g of dried oregano plants (OPs) to provide daily dosage of 1 ml of essential oil (EO) per animal. The experimental period lasted 69 days and individual rumen fluid samples were obtained every 2 weeks at 0 and 4 hr after feeding. The results showed that dietary supplementation with OPs increased the protease activity (p < .001) and ammonia concentration (p < .05) in the rumen. Among the studied microbial populations, Peptostreptococcus anaerobius (p = .028) and Clostridium sticklandii (p < .001) were found to be the most sensitive to oregano at the current dosage. Furthermore, the total methanogen population significantly decreased (p < .05). It is concluded that a long‐term dietary administration of OPs can suppress specific rumen micro‐organisms and modify rumen fermentation favourably at least by means of suppressing methanogens.     

Detection of 11 beta‐hydroxysteroid dehydrogenase type 1, the glucocorticoid and mineralocorticoid receptor in various adipose tissue depots of dairy cows supplemented with conjugated linoleic acids

Early lactating cows mobilize adipose tissue (AT) to provide energy for milk yield and maintenance and are susceptible to metabolic disorders and impaired immune response. Conjugated linoleic acids (CLA), mainly the trans‐10, cis‐12 isomer, reduce milk fat synthesis and may attenuate negative energy balance. Circulating glucocorticoids (GC) are increased during parturition in dairy cows and mediate differentiating and anti‐inflammatory effects via glucocorticoid (GR) and mineralocorticoid receptors (MR) in the presence of the enzyme 11β‐hydroxysteroid dehydrogenase type 1 (11βHSD1). Activated GC are the main ligands for both receptors in AT; therefore, we hypothesized that tissue‐specific GC metabolism is effected by varying amounts of GR, MR and 11βHSD1 and/or their localization within AT depots. Furthermore, the lipolytic and antilipogenic effects of CLA might influence the GC/GR/MR system in AT. Therefore, we aimed to localize GR and MR as well as the expression pattern and activity of 11βHSD1 in different AT depots during early lactation in dairy cows and to identify potential effects of CLA. Primiparous German Holstein cows were divided into a control (CON) and a CLA group. From day 1 post‐partum (p.p.) until sample collection, the CLA group was fed with 100 g/d CLA (contains 10 g each of the cis‐9, trans‐11 and the trans‐10, cis‐12‐CLA isomers). CON cows (n = 5 each) were slaughtered on day 1, 42 and 105 p.p., while CLA cows (n = 5 each) were slaughtered on day 42 and 105 p.p. Subcutaneous fat from tailhead, withers and sternum, and visceral fat from omental, mesenteric and retroperitoneal depots were sampled. The localization of GR and 11βHSD1 in mature adipocytes – being already differentiated – indicates that GC promote other effects via GR than differentiation. Moreover, MR were observed in the stromal vascular cell fraction and positively related to the pre‐adipocyte marker Pref‐1. However, only marginal CLA effects were observed in this study.     

Papaya (Carica papaya) leaf methanolic extract modulates in vitro rumen methanogenesis and rumen biohydrogenation

Papaya leaf methanolic extract (PLE) at concentrations of 0 (CON), 5 (LLE), 10 (MLE) and 15 (HLE) mg/250 mg dry matter (DM) with 30 mL buffered rumen fluid were incubated for 24 h to identify its effect on in vitro ruminal methanogenesis and ruminal biohydrogenation (BH). Total gas production was not affected (P > 0.05) by addition of PLE compared to the CON at 24 h of incubation. Methane (CH₄) production (mL/250 mg DM) decreased (P < 0.05) with increasing levels of PLE. Acetate to propionate ratio was lower (P <0.05) in MLE (2.02) and HLE (1.93) compared to the CON (2.28). Supplementation of the diet with PLE significantly (P <0.05) decreased the rate of BH of C18:1n‐9 (oleic acid; OA), C18:2n‐6 (linoleic acid; LA), C18:3n‐3 (linolenic acid; LNA) and C18 polyunsaturated fatty acids (PUFA) compared to CON after 24 h incubation, which resulted in higher concentrations of BH intermediates such as C18:1 t11 (vaccenic acid; VA), c9t11 conjugated LA (CLA) (rumenic acid; RA) and t10c12 CLA. Real‐time PCR analysis indicated that the total bacteria, total protozoa, Butyrivibrio fibrisolvens and methanogen population in HLE decreased (P <0.05) compared to CON, but the total bacteria and B. fibrisolvens population were higher (P < 0.05) in CON compared to the PLE treatment groups.     

Effects of supplementing an active dry yeast product on rumen microbial community composition and on subsequent rumen fermentation of lactating cows in the mid‐to‐late lactation period

The effects of supplementing feed of cows in mid‐to‐late lactation with an active yeast product (Actisaf Sc 47) were evaluated using 15 Holstein cows in a replicated 3 × 3 Latin square design. The animals were fed a mixed ration with 33% neutral detergent fiber, consisting of timothy hay (29.8%), a commercial concentrate (70.0%) and commercial calcium triphosphate (0.2%), twice daily to meet 105% of their energy requirement. Yeast supplement was set at 0, 5 and 10 g per day over 21‐day periods, each of which consisted of 14 days for adaptation followed by 7 days of data collection. Milking performance, plasma metabolite parameters, rumen volatile fatty acids, lipopolysaccharide and microbial properties were measured. Although there were no significant differences in feeding and milking performance or blood parameters associated with supplementation, the acetate to propionate ratio in the rumen fluid tended to decrease (P = 0.08). The population of Bacteroidetes tended to be less prominent (P = 0.07) and the fibrolytic bacterium Fibrobacter significantly increased (P < 0.05) in the rumen fluid of the yeast 10 g group compared with that of the control. These data suggest that effects of supplementing live yeast to cows in mid‐to‐late lactation may be limited to microbial composition and fermentation characteristics in the rumen.     

Feed degradability,rumen fermentation and blood metabolites in response to essential oil addition to fistulated non‐lactating dairy cow diets

The effects of essential oils (EOs) on ruminal nutrient disappearance, rumen fermentation and blood metabolites in fistulated non‐lactating dairy cows were studied. Four fistulated non‐lactaing dairy cows were used in a 4 × 4 Latin square design; the experiment consisted of four periods of 21 days in each period, with the first 14 days for adaptation followed by 7 days of measurement period. Animals were fed 3 kg/day of 21% crude protein (CP) concentrate and ad libitum corn silage. Treatments were: (i) control; (ii) 2 mL Allicin/cow/day; (iii) 2 mL zingiberene/cow/day; and (iv) 2 mL citral/cow/day. The results demonstrated that EOs increased dry matter and neutral detergent fiber degradabilities at 48 and 72 h, but had no effect on acid detergent fiber and CP degradabilities. EOs did not change ruminal pH, ammonia nitrogen, protozoa, volatile fatty acid concentrations and blood glucose but reduced blood urea nitrogen at 4 h.     

Induction of a transient acidosis in the rumen simulation technique

Feeding high concentrate diets to cattle results in an enhanced production of short‐chain fatty acids by the micro‐organisms in the rumen. Excessive fermentation might result in subclinical or clinical rumen acidosis, characterized by low pH, alterations in the microbial community and lactate production. Here, we provide an in vitro model of a severe rumen acidosis. A transient acidosis was induced in the rumen simulation technique by lowering bicarbonate, dihydrogen phosphate and hydrogen phosphate concentrations in the artificial saliva while providing a concentrate‐to‐forage ratio of 70:30. The experiment consisted of an equilibration period of 7 days, a first control period of 5 days, the acidosis period of 5 days and a second control period of 5 days. During acidosis induction, pH decreased stepwise until it ranged below 5.0 at the last day of acidosis (day 17). This was accompanied by an increase in lactate production reaching 11.3 mm at day 17. The daily production of acetate, propionate and butyrate was reduced at the end of the acidosis period. Gas production (methane and carbon dioxide) and NH₃‐N concentration reached a minimum 2 days after terminating the acidosis challenge. While the initial pH was already restored 1 day after acidosis, alterations in the mentioned fermentation parameters lasted longer. However, by the end of the experiment, all parameters had recovered. An acidosis‐induced alteration in the microbial community of bacteria and archaea was revealed by single‐strand conformation polymorphism. For bacteria, the pre‐acidotic community could be re‐established within 5 days, however, not for archaea. This study provides an in vitro model for a transient rumen acidosis including biochemical and microbial changes, which might be used for testing feeding strategies or feed additives influencing rumen acidosis.     

Provision of beta‐glucan prebiotics (cellooligosaccharides and kraft pulp) to calves from pre‐ to post‐weaning period on pasture

We conducted two feeding experiments to evaluate the effects of supplementation with either cellooligosaccharide or kraft pulp on growth performance in grazing beef calves (Japanese Black) from 4 weeks pre‐weaning to 12 to 16 weeks post‐weaning. In Experiment 1 (20‐week duration), nine calves (2.9‐month‐old females) were assigned to either a control group (CON) or an experimental group (CEL) fed cellooligosaccharide at a rate of 10 g/day mixed with concentrate. Average daily weight gain tended to be greater in CEL than in CON, especially after 1 month of weaning. In Experiment 2 (16‐week duration), 10 calves (2.0‐month‐old females) were assigned to either a control group or an experimental group (KRA) fed kraft pulp at a rate of 10% replacement of total digestible nutrients with concentrate. The proportion of fibrolytic bacteria increased and that of methanogenic Archaea decreased in the rumen microbial community composition of KRA calves in Experiment 2, whereas the decrease in Fibrobacter and Archaea was observed in CEL calves at first 4 weeks in Experiment 1. We conclude that beta‐glucan prebiotic supplementation to grazing calves at pre‐weaning would affect rumen microbial composition and modified rumen fermentation characteristics, leading to a better rumen environment via different means.     

Effects of isobutyrate supplementation on ruminal microflora,rumen enzyme activities and methane emissions in Simmental steers

The objective of this study was to evaluate the effects of isobutyrate supplementation on rumen microflora, enzyme activities and methane emissions in Simmental steers consuming a corn stover‐based diet. Eight ruminally cannulated Simmental steers were used in a replicated 4 × 4 Latin square experiment. The treatments were control (without isobutyrate), low isobutyrate (LIB), moderate isobutyrate (MIB) and high isobutyrate (HIB) with 8.4, 16.8 and 25.2 g isobutyrate per steer per day respectively. Isobutyrate was hand‐mixed into the concentrate portion. Diet consisted of 60% corn stover and 40% concentrate [dry matter (DM) basis]. Dry matter intake (averaged 9 kg/day) was restricted to a maximum of 90% of ad libitum intake. Population of total bacteria, cellulolytic bacteria and anaerobic fungi were linearly increased, whereas that of protozoa and total methanogens was linearly reduced with increasing isobutyrate supplementation. Real‐time PCR quantification of population of Ruminococcus albus, Ruminococcus flavefaciens, Butyrivibrio fibrisolvens and Fibrobacter succinogenes was linearly increased with increasing isobutyrate supplementation. Activities of carboxymethyl cellulase, xylanase and β‐glucosidase were linearly increased, whereas that of protease was linearly reduced. Methane production was linearly decreased with increasing isobutyrate supplementation. Effective degradabilities of cellulose and hemicellulose of corn stover were linearly increased, whereas that of crude protein in diet was linearly decreased with increasing isobutyrate supplementation. The present results indicate that isobutyrate supplemented improved microflora, rumen enzyme activities and methane emissions in steers. It was suggested that the isobutyrate stimulated the digestive micro‐organisms or enzymes in a dose‐dependent manner. In the experimental conditions of this trial, the optimum isobutyrate dose was approximately 16.8 g isobutyrate per steer per day.     

Effect of pre‐ and post‐partum supplementation with lipid‐encapsulated conjugated linoleic acid on milk yield and metabolic status in multiparous high‐producing dairy cows

We evaluated the lactation performance, liver lipid content and plasma metabolites indicating the energy balance of dairy cows supplemented with conjugated linoleic acid (CLA) pre‐ and post‐partum (PP) vs. only PP. A total of 60 cows were divided into three groups (n = 20). Daily diet of cows was supplemented with 14 g of CLA (7 g cis‐9, trans‐11 and 7 g trans‐10, cis‐12 isomers) from week 3 before the expected date of calving (group CLA1), or from the day of calving (group CLA2) until 77–91 days PP. Control cows were fed an isocaloric, isonitrogenous and isolipidic diet without CLA. Between week 3 and week 6 PP, the milk yield of cows in both CLA‐treated groups was approximately 4.5 kg higher (p < 0.05) than in control. Milk fat concentrations decreased from week 3 and were lower in both CLA groups than in control (p < 0.01). Body condition score loss was lower (p < 0.05) in the CLA1 than in the control group on week 5 PP. By week 11 PP, the body condition of both CLA1 and CLA2 groups exceeded that of control. Plasma non‐esterified fatty acid was lower in CLA1 compared to CLA2 and control during the early PP period (p < 0.05), while this difference faded away by the late PP period. Beta‐hydroxybutyrate (BHBA) increased rapidly in all groups following calving. In CLA1 group, it began to decrease sooner than in CLA2 and control. The prevalence of subclinical ketosis (BHBA > 1.2 mm ) was lower in CLA1 group than in CLA2 and control (p < 0.05). Liver biopsy analyses showed that CLA1 treatment decreased (p < 0.05) the total lipid content of liver compared to control at week 5 after calving. Our results show that CLA supplementation is more efficient in alleviating body mass mobilization and decreasing the incidence of subclinical ketosis when applied as early as 3 weeks before calving than started feeding after calving.     

Inflammatory and metabolic responses to an intramammary lipopolysaccharide challenge in early lactating cows supplemented with conjugated linoleic acid

Supplementation of dairy cows with trans‐10, cis‐12 conjugated linoleic acid (CLA) allows nutrient repartitioning despite an energy deficiency in early lactation, which might be a benefit for the immune system, too. In this study, we investigated potential nutrient sparing effects of CLA in early lactating cows with low plasma glucose concentrations exposed to an intramammary lipopolysaccharide (LPS) challenge. Fifteen multiparous Holstein cows were exposed to an intramammary LPS challenge in week 4 p.p. Eight cows (CLA) were supplemented daily with 70 g of lipid‐encapsulated CLA (6.8 g trans‐10, cis‐12 and 6.6 g of the cis‐9, trans‐11 CLA isomer; CLA) and seven cows with 56 g of control fat (CON). Blood samples were obtained every 30 min along with rectal temperature, heart and respiratory rate, and milk samples were taken hourly until 10 hr after the LPS application. Plasma was analysed for concentrations of glucose, free fatty acids, beta‐hydroxybutyrate (BHB), cortisol, insulin and glucagon. In milk, somatic cell count and activity of lactate dehydrogenase were determined. Initial plasma glucose concentration was lower in CLA than in CON. During the immunostimulation, CLA had higher glucose concentrations than CON, and BHB decreased distinctly in CLA, whereas CON cows maintained BHB concentration at a lower level. Body temperature in CLA increased earlier, the difference between peak and basal temperature was higher, and the decline thereafter occurred earlier. In conclusion, CLA supplementation of early lactating cows exposed to an intramammary LPS challenge affected local and systemic immune responses. We assume that CLA supplementation triggered glycogen storage. Cows supplemented with CLA provided more glucose and preferentially used BHB as an energy source during the immune response. The more intense metabolic and more concentrated endocrine responses support an immunomodulatory effect of CLA supplementation.     

Efficacy of Megasphaera elsdenii inoculation in subacute ruminal acidosis in cattle

Two consecutive experiments were carried out to determine efficacy of Megasphaera elsdenii inoculation in alleviation of subacute ruminal acidosis (SARA). In the first experiment, SARA was induced by feeding corn‐ and wheat‐based diets (20%, 40%, 60% and 80% of TMR, DM basis) in six ruminally cannulated heifers. Continuous pH was obtained using data loggers embedded in rumen. In corn (80%)‐ and wheat (60%)‐based diets ruminal pH ranged from 5.2 to 5.6 for 7.77 and 5.93 hr. In the second experiment (5 day), M. elsdenii (200 ml; 2.4 x 10¹⁰ cfu/ml) was inoculated during the first two days. During the SARA induction period, M. elsdenii and S. bovis in rumen liquor were more abundant in wheat‐based feeding (7.97 and 8.77) than in corn‐based feeding (7.06 and 7.95 per ml, log basis; p < 0.0001 for both). M. elsdenii inoculation increased total volatile fatty acids (VFA) concentration when corn‐based diet was fed, whereas it decreased total VFA concentration when wheat‐based diet was fed (p < 0.004). There was a decrease in the propionic acid proportion (24.04%–19.08%; p < 0.002), whereas no alteration in lactate and ammonia concentrations was observed. M. elsdenii inoculation increased protozoa count (from 5.39 to 5.55 per ml, log basis; p < 0.009) and decreased S. bovis count (from 9.18 to 7.95 per ml, log basis; p < 0.0001). The results suggest that M. elsdenii inoculation may help prevent SARA depending on dietary grain through altering rumen flora as reflected by a decrease in S. bovis count and an increase in protozoa count.     

Effects of fumaric acid on rumen fermentation,milk composition and metabolic parameters in lactating cows

The aim of this study was to determine the influence of fumaric acid (FA) on ruminal fermentation and its effects on the acid‐base balance of seven ruminally and duodenally fistulated multiparous German Holstein cows. The experiment was conducted in a change‐over design with three periods in which the animals were randomly arranged in one of three treatments: Control (C; without FA), 300 or 600 g FA per day. The diets consisted of 7.4 kg DM grass silage, 4.2 kg concentrate mixture and 0, 300 or 600 g FA or wheat starch as isocaloric compensation per day and cow. FA supplementation decreased the rumen pH, acetic acid and butyric acid and increased propionic acid in rumen fluid. The results of the single‐strand conformation polymorphism analysis (SSCP) did not show an influence of FA on the microbial population in the rumen. The beta‐hydroxybutyrate (BHB) concentration in blood and the pH of the urine decreased, while the blood gases were unaffected by supplementation of the acid. The microbial protein per MJ ME decreased in the duodenum with FA supplementation. The milk fat concentration decreased after addition of FA. We conclude that in this study feeding of up to 600 g FA per day did not result in an acidosis. It seems that up to 600 g FA per day did not have a significant influence on the acid‐base balance of dairy cows.     

Effects of rumen‐protected methionine on milk production in early lactation dairy cattle fed with a diet containing 14.5% crude protein

We evaluated the influence on milk production of feeding early lactation cows a diet that included 14.5% crude protein (CP) and that did not meet methionine (Met) requirements or that met them by supplying rumen‐protected Met (RPMet). Thirty‐nine multiparous Holstein cows were allocated into two groups. For 15 weeks after calving, each group was fed one of the two total mixed rations, Control (n = 20) or Treatment (n = 19). The Treatment group received added RPMet at 0.034% (8 g/day) of the Control diet on dry matter basis. The adequacies of Met for the Control and Treatment groups were 96% and 106%, respectively, and for other amino acids, >110%. The CP level (14.5%) was 1 percentage point lower than that recommended by the Japanese Feeding Standard (2006). No between‐group differences were found in milk yield (40 kg/day), milk composition, plasma profile, rumen fermentation, nitrogen balance, or cow health. Met intake and the amount of rumen‐undegradable feed Met were higher in the Treatment group (p < 0.05). Microbial Met and total metabolizable Met did not differ between groups. Supplying RPMet in a 14.5% CP diet during early lactation did not dramatically affect milk production, because the amount of total metabolizable Met was unchanged.     

Concentrations of retinol and tocopherols in the milk of cows supplemented with conjugated linoleic acid

This study was performed to investigate the hypothesis that supplementation of conjugated linoleic acid (CLA) changes the concentrations of retinol and tocopherols in the milk of cows. To investigate this hypothesis, Holstein cows received daily from 3 weeks ante‐partum to 14 weeks post‐partum either 172 g of a CLA‐free rumen‐protected control fat (control group, n = 20) or the same amount of a rumen‐protected CLA fat, supplying 4.3 g of cis‐9, trans‐11 CLA and 3.8 g of trans‐10, cis‐12 CLA per d (CLA group, n = 20). Milk samples (collected at weeks 1, 3, 5, 8 and 11 of lactation) were analysed for retinol, α‐ and γ‐tocopherol concentrations. Milk of cows supplemented with CLA had higher concentrations of retinol (+34%), α‐tocopherol (+44%) and γ‐tocopherol (+21%) than milk of control cows (p < 0.05). The daily output of these vitamins via milk was also greater in cows of the CLA group than in cows of the control group (+36, 50 and 24% for retinol, α‐tocopherol and γ‐tocopherol, respectively, p < 0.05). In agreement with higher concentrations of tocopherols, concentrations of thiobarbituric acid‐reactive substances, determined in milk of week 5, were lower in cows of the CLA group than in control cows, indicative of a lower susceptibility of milk lipids to peroxidation. Plasma concentrations of retinol and α‐tocopherol, determined at 1 and 5 weeks post‐partum, were not different between the two groups of cows. In conclusion, this study shows that supplementing dairy cows with a moderate amount of CLA causes an increase of the concentrations of vitamins A and E in the milk and results in an increased output of those vitamins via milk. These effects might be beneficial with respect to the nutritional value of dairy products and the susceptibility of milk fat to oxidative deterioration.     

Effects of Fusarium mycotoxins in rations with different concentrate proportions on serum haptoglobin and hepatocellular integrity in lactating dairy cows

It was hypothesized that long‐term intake of a diet contaminated with deoxynivalenol (DON) and differing in the proportion of concentrate might affect hepatocellular integrity and function as well as biomarkers of systemic inflammation in lactating dairy cows. In Period 1 (11 weeks), 26 lactating German Holstein cows (13 primiparous and 13 pluriparous, 31 days in milk, 522 kg body weight, on average) were divided into two groups and fed diets (50% concentrate) with (MYC, n = 12; on average 5.3 mg DON/kg DM) or without (CON, n = 14) DON contaminations. In Period 2 (16 weeks), each group was further divided into two groups to test whether elevated concentrate proportion as additional burden might enhance the toxicity of DON. The cows in MYC60 (n = 6; 4.6 mg DON/kg DM) and CON60 (n = 7) received the diet with 60% concentrate, while cows in MYC30 (n = 6; 4.4 mg DON/kg DM) and CON30 (n = 7) received the diet with 30% concentrate. Blood samples were taken in biweekly intervals for activities of aspartate amino transferase (AST), glutamate dehydrogenase (GLDH) and gamma‐glutamyl transferase as well as for concentration of total bilirubin and haptoglobin. Biopsies from liver were collected in week 27 for morphological analyses. No DON effect was found for the variables assessed in blood. The diet with 60% concentrate led to higher activities of AST and GLDH in Period 2. No morphological change was found by both light and electron microscopic analyses of liver samples. Results indicated that long‐term intake of DON‐contaminated diet over 27 weeks led to neither relevant damages of hepatocytes nor systemic inflammatory responses in lactating dairy cows, even if the dietary concentrate proportion was increased to 60%.