A 2-year study reveals implications of feeding management and exposure to mycotoxins on udder health, performance, and fertility in dairy herds.

We recently reported the ubiquitous occurrence of mycotoxins and their secondary metabolites in dairy rations and a substantial variation in the feeding management among Austrian dairy farms. The present study aimed to characterize to which extent these factors contribute to the fertility, udder health traits, and performance of dairy herds. During 2019 and 2020, we surveyed 100 dairy farms, visiting each farm 2 times and collecting data and feed samples. Data collection involved information on the main feed ingredients, nutrient composition, and the levels of mycotoxin and other metabolites in the diet. The annual fertility and milk data of the herds were obtained from the national reporting agency. Calving interval was the target criterion for fertility performance, whereas the percentage of primiparous and multiparous cows in the herd with somatic cell counts above 200,000 cells/mL was the criterion for impaired udder health. For each criterion, herds were classified into 3 groups: high/long, mid, and low/short, with the cut-off corresponding to the <25th and >75th percentiles and the rest of the data, respectively. Accordingly, for the calving interval, the cut-offs for the long and short groups were ≥400 and ≤380 d, for the udder health in primiparous cows were ≥20% and ≤8% of the herd, and for the udder health in multiparous cows were ≥35% and ≤20% of the herd, respectively. Quantitative approaches were further performed to define potential risk factors in the herds. The high somatic cell count group had higher dietary exposure to enniatins (2.8 vs. 1.62 mg/cow per d), deoxynivalenol (4.91 vs. 2.3 mg/cow per d), culmorin (9.48 vs. 5.72 mg/cow per d), beauvericin (0.32 vs. 0.18 mg/cow per d), and siccanol (13.3 vs. 5.15 mg/cow per d), and total Fusarium metabolites (42.8 vs. 23.2 mg/cow per d) and used more corn silage in the ration (26.9% vs. 17.3% diet DM) compared with the low counterparts. Beauvericin was the most substantial contributing variable among the Fusarium metabolites, as indicated by logistic regression and modeling analyses. Logistic analysis indicated that herds with high proportions of cows with milk fat-to-protein ratio >1.5 had an increased odds for a longer calving interval, which was found to be significant for primiparous cows (odds ratio = 5.5, 95% confidence interval = 1.65-21.7). As well, herds with high proportions of multiparous cows showing levels of milk urea nitrogen >30 mg/dL had an increased odds for longer calving intervals (odds ratio = 2.96, 95% confidence interval = 1.22-7.87). In conclusion, the present findings suggest that dietary contamination of Fusarium mycotoxins (especially emerging ones), likely due to increased use of corn silage in the diet, seems to be a risk factor for impairing the udder health of primiparous cows. Mismatching dietary energy and protein supply of multiparous cows contributed to reduced herd fertility performance.

Adaptive responses in short-chain fatty acid absorption, gene expression, and bacterial community of the bovine rumen epithelium recovered from a continuous or transient high-grain feeding.

The feeding of high-grain diets to dairy cows commonly results in lowered pH and ruminal dysbiosis, characterized by changes in absorption dynamics of short-chain fatty acids (SCFA) across the reticuloruminal wall, epithelial function, and the epithelial bacteria community structure. Therefore, the present study evaluated the effect of high-grain feeding persistence on the absorption kinetics of reticuloruminal SCFA, gene expression in the rumen epithelium, and the associated shifts in the epithelial bacteria in cows recovering from either a long-term continuous high-grain feeding model or a long-term transient high-grain feeding model. In a crossover study design, 8 nonlactating Holstein cows were fed 60% concentrate either continuously for 4 wk (continuous) or with a 1-wk break in the second week of the high-grain feeding (transient). After the high-grain feeding, all animals were fed a diet of 100% forage (recovery) for an additional 8 wk. Rumen papilla biopsies and SCFA absorption measurements were taken at the start of the trial (baseline), after the 4-wk high-grain feeding (49 d), after 2-wk recovery forage feeding (63 d), and after 8-wk recovery forage (105 d). Absorption of SCFA was determined in vivo using the washed and isolated reticulorumen technique. Rumen papillae biopsies were used for adherent bacterial DNA and host RNA extraction. The epithelial bacteria were determined using Illumina MiSeq (Microsynth AG, Balgach, Switzerland) sequencing of the 16S rRNA gene. No significant effects of the high-grain feeding model were seen for bacterial diversity. However, bacterial diversity increased with time spent in the recovery forage feeding period regardless of feeding model. The relative abundance of Acidobacteria phyla and Acetivibrio spp. increased when animals were fed a transient high-grain feeding model. A trend toward increased CLDN4 expression was observed in the continuous model. Furthermore, there were interactions between feeding model and sampling day for gene targets CD14, DRA, NHE2, NHE3, and MCT2. When comparing length of recovery, in the continuous model increased relative absorption of SCFA was sustained at 63 d but dropped to baseline measurements at 105 d. A similar pattern was found with the transient model but it did not reach significance. The only gene target that was found to significantly correlate to relative absorption of SCFA was DRA (correlation coefficient ≤ -0.41). Whereas, genera Alkalibaculum, Anaerorhabdus, Coprococcus, and Dethiobacter all showed positive correlations to gene targets for pH regulation (NHE2 and NHE3) and SCFA uptake (MCT1) but negative correlations to SCFA absorption. We conclude that while the rumen absorption and epithelial bacteria were able to recover to baseline levels after 8 wk of forage feeding, the time needed for re-establishment of homeostasis in host gene expression is longer, especially when high-grain feeding is interrupted.

Metabolic and stress responses in dairy cows fed a concentrate-rich diet and submitted to intramammary lipopolysaccharide challenge.

Feeding dairy cows diets rich in grain often leads to subacute rumen acidosis (SARA), which might affect their responsiveness to immunogenic stimuli such as exogenous lipopolysaccharide (LPS), and can lead to metabolic alterations. The main objective of this study was to investigate if SARA affects the stress and metabolic health responses resulting from an intramammary LPS challenge. Before the intramammary LPS challenge, the SARA cows showed higher blood glucose and a tendency for higher lactate and aspartate aminotransferase as well as a trend toward lower ß-hydroxybutyrate (BHBA) and γ-glutamyltransferase compared with control cows. After the LPS challenge, the serum cortisol concentration markedly increased and the calcium concentration decreased both in SARA and control cows. In SARA-LPS cows, however, the lactate concentration increased due to the LPS infusion, whereas it remained unchanged in the control cows. A lower serum BHBA concentration was found in SARA-LPS compared with control-LPS cows. Higher non-esterified fatty acid concentrations were found in control-LPS cows shortly before the LPS challenge compared with SARA cows, challenged or not with LPS, whereas it did not differ from SARA-LPS cows thereafter. In conclusion, the results suggest that intramammary LPS challenge induced stress and lowered calcium concentration in all dairy cows, whereby this challenge showed lower BHBA and higher lactate levels in cows with SARA conditions.

Signals for identifying cows at risk of subacute ruminal acidosis in dairy veterinary practice.

Controlling rumen disorders is critical to ensure successful dairy herd health management. Lactation diets of dairy cows are commonly rich in concentrates and low in physically effective fibre. Feeding of these diets increases the risk of rumen disorders with far-reaching consequences for cattle health, welfare and sustainability of dairy production. The term subacute ruminal acidosis or SARA is often used as a synonym for poor rumen health. Being subclinical, SARA lacks of clear symptoms and is therefore difficult to diagnose and to control in the practice. This review article summarises common and identifies new direct and indirect cow signals related to SARA. We have performed a scientific evaluation and interpretation of each of these cow signals by highlighting their advantages and disadvantages from the practitioner's point of view. The gold standard of SARA cow signals still remains direct measurement of ruminal pH. However, continuous pH monitoring is cost-intensive and often biased by sensor drift. Single-point ruminal pH measurements by oral stomach tubing or rumenocentesis have strong limitations. Therefore, there is a need for reliable and robust markers of SARA that are easily accessible and inexpensive. Such indirect parameters are the observation of chewing and feeding activities, as well as the monitoring of milk, faecal, urine and blood variables. Also, novel technologies that allow rapid and non-invasive measurement of the rumen mucosa thickness and ruminal motility patterns might provide advantages in SARA diagnosis. Due to several constraints of these indirect diagnostic tools, such as limited specificity and sensitivity, we strongly recommend using a combination of the signals to reliably identify cows at risk of SARA in a dairy herd.

Restoration of in situ fiber degradation and the role of fibrolytic microbes and ruminal pH in cows fed grain-rich diets transiently or continuously.

In this study, we used two different grain-rich feeding models (continuous or transient) to determine their effects on in situ fiber degradation and abundances of important rumen fibrolytic microbes in the rumen. The role of the magnitude of ruminal pH drop during grain feeding in the fiber degradation was also determined. The study was performed in eight rumen-fistulated dry cows. They were fed forage-only diet (baseline), and then challenged with a 60% concentrate diet for 4 weeks, either continuously (n=4 cows) or transiently (n=4 cows). The cows of transient feeding had 1 week off concentrate in between. Ruminal degradation of grass silage and fiber-rich hay was determined by the in situ technique, and microbial abundances attached to incubated samples were analyzed by quantitative PCR. The in situ trials were performed at the baseline and in the 1st and the last week of concentrate feeding in the continuous model. The in situ trials were done in cows of the transient model at the baseline and in the 1st week of the re-challenge with concentrate. In situ degradation of NDF and ADF of the forage samples, and microbial abundances were determined at 0, 4, 8, 24 and 48 h of the incubation. Ruminal pH and temperature during the incubation were recorded using indwelling pH sensors. Compared with the respective baseline, both grain-rich feeding models lowered ruminal pH and increased the duration of pH below 5.5 and 5.8. Results of the grass silage incubation showed that in the continuous model the extent of NDF and ADF degradation was lower in the 1st, but not in the last week compared with the baseline. For the transient model, degradation of NDF of the silage was lower during the re-challenge compared with the baseline. Degradation of NDF and ADF of the hay was suppressed by both feeding models compared with the respective baseline. Changes in fiber degradation of either grass silage or hay were not related to the magnitude of ruminal pH depression during grain-rich feeding. In both feeding models total fungal numbers and relative abundance of Butyrivibrio fibrisolvens attached to the incubated forages were decreased by the challenge. Overall, Fibrobacter succinogenes was more sensitive to the grain challenge compared with Ruminococcus albus and Ruminococcus flavefaciens. The study provided evidence for a restored ruminal fiber degradation after prolonged time of grain-rich feeding, however depending on physical and chemical characteristics of forages.

Intramammary infusion of Escherichia coli lipopolysaccharide negatively affects feed intake, chewing, and clinical variables, but some effects are stronger in cows experiencing subacute rumen acidosis.

Feeding high-grain diets increases the risk of subacute rumen acidosis (SARA) and adversely affects rumen health. This condition might impair the responsiveness of cows when they are exposed to external infectious stimuli such as lipopolysaccharide (LPS). The main objective of this study was to evaluate various responses to intramammary LPS infusion in healthy dairy cows and those experimentally subjected to SARA. Eighteen early-lactating Simmental cows were subjected to SARA (n = 12) or control (CON; n = 6) feeding conditions. Cows of the control group received a diet containing 40% concentrates (DM basis) throughout the experiment. The intermittent SARA feeding regimen consisted in feeding the cows a ration with 60% concentrate (DM basis) for 32 d, consisting of a first SARA induction for 8 d, switched to the CON diet for 7 d, and re-induction during the last 17 d. On d 30 of the experiment, 6 SARA (SARA-LPS) and 6 CON (CON-LPS) cows were intramammary challenged once with a single dose of 50 µg of LPS from Escherichia coli (O26:B6), whereas the other 6 SARA cows (SARA-PLA) received 10 mL of sterile saline solution as placebo. To confirm the induction of SARA, the reticular pH was continuously monitored via wireless pH probes. The DMI remained unchanged between SARA and CON cows during the feeding experiment, but was reduced in both treatment groups receiving the LPS infusion compared with SARA-PLA, whereby a significant decline was observed for cows of the SARA-LPS treatment (-38%) compared with CON-LPS (-19%). The LPS infusion did not affect the reticuloruminal pH dynamics, but significantly enhanced ruminal temperature and negatively affected chewing behavior. The ruminal temperature increased after the LPS infusion and peaked about 1 h earlier in SARA-LPS cows compared with the cows of the CON-LPS treatment. Moreover, a significant decline in milk yield was found in SARA-LPS compared with CON-LPS following the LPS infusion. Cows receiving LPS had elevated somatic cell counts, protein, and fat contents in milk as well as decreased lactose contents and pH following the LPS infusion, whereby the changes in milk constituents were more pronounced in SARA-LPS than CON-LPS cows. Rectal temperature and pulse rate were highest 6 h after LPS infusion, but rumen contractions were not affected by the LPS infusion. The data suggest that a single intramammary LPS infusion induced fever and negatively affected feed intake, chewing activity, rectal temperature, and milk yield and composition, whereby these effects were more pronounced in SARA cows.

Transient feeding of a concentrate-rich diet increases the severity of subacute ruminal acidosis in dairy cattle.

The objective of this study was to investigate the effect of the pattern of concentrate-rich feeding on subacute ruminal acidosis (SARA), its severity, and the corresponding changes in VFA concentration. Eight rumen-cannulated Holstein cows were assigned to a 2 × 2 crossover design with 2 SARA challenge models and 2 experimental runs ( = 8 per treatment). Each run lasted for 40 d, consisting of a 6-d baseline, a 6-d gradual grain adaptation, and a 28-d SARA challenge period. The 2 SARA challenge models were transient (TRA) and persistent (PER) SARA. Initially, all cows were subjected to a forage-only diet (baseline) and gradually switched to 60% concentrate (DM basis). Then, cows in the PER model were continuously challenged for 28 d, whereas cows in the TRA model had a 7-d break from the SARA diet and were fed the forage-only diet after the first 7 d of SARA challenge. Thereafter, the TRA cows were rechallenged with the SARA diet. Wireless ruminal pH sensors were used to obtain ruminal pH profiles and temperature over the experimental period. For the determination of VFA, free ruminal liquid (FRL) and particle-associated ruminal liquid (PARL) were collected once for the baseline and twice (d 20 and 40 for the PER model) or 3 times (d 13, 30, and 40 for the TRA model) during SARA, each time at 0, 4, and 8 h after the morning feeding. Cows in both models experienced SARA albeit with day-to-day variation. From the start until the first 7-d SARA, cows of both models had similar pH profiles, but during the rechallenge, SARA was more severe in the TRA model than in the PER model based on lower daily mean ruminal pH (5.93 vs. 6.15; SEM 0.058) and double the amount of time at pH < 5.8 (497 vs. 278 min; SEM 68.61, < 0.05). Mean ruminal temperature was raised during SARA compared with the baseline (38.9 vs. 38.7°C; SEM 0.057, < 0.001). Concentrations of VFA increased with increasing time after feeding ( < 0.001). In general, SARA challenge (d 40 vs. the baseline), but not the challenge model, altered VFA concentrations and profile of both FRL and PARL by increasing the amounts of propionate and butyrate, whereas total VFA concentration was less affected. Proportions of VFA shifted over the duration of SARA challenge with more propionate but less acetate and butyrate proportions with advancing days of SARA challenge, leading to the values of the last SARA day being different from the earlier days ( < 0.05). In conclusion, the TRA condition led to the higher severity of SARA, but factors beyond feed intake and VFA alterations seemed to play a role.

Apparent recovery of C18 polyunsaturated fatty acids from feed in cow milk: a meta-analysis of the importance of dietary fatty acids and feeding regimens in diets without fat supplementation.

A meta-analysis was conducted using the results of 82 experiments (78 publications, 266 treatments) to investigate the importance of dietary C18 fatty acids (FA) and feeding regimen for milk C18 FA profile and apparent recovery of selected FA relative to intake of these FA or their precursors. Feeding treatments based on lipid-supplemented diets were excluded. Feeding regimens were defined as grazing [including partial and full-time grazing, at dietary concentrate proportions from 0 to 44% dry matter (DM)], forage-based indoor feeding [>65% forage of total DM intake (DMI)], and concentrate-based indoor feeding (forage DMI ≤65% of DMI). Linoleic acid (LLA), α-linolenic acid (ALA), and total C18 FA proportions in milk fat increased linearly with the respective dietary FA content in all feeding regimens tested. This effect was highest in the forage-based indoor feeding. Slopes were lowest for the grazing regimens, especially regarding ALA and the sum of all C18 FA, whereas the intercepts of the prediction equations of milk ALA and total C18 FA proportions were highest for grazing regimens. This indicates that, in grazing cows, factors other than dietary FA contents determine the C18 FA composition of the milk fat. At equal dietary LLA contents, the type of feeding regimen showed no significant effect on LLA proportion in milk fat. Milk fat proportions of rumenic acid and vaccenic acid were positively related to the sum of dietary ALA and LLA contents. Grazing regimens led to the strongest enrichment of rumenic acid and vaccenic acid in milk fat. The apparent recovery of ALA, LLA, and total C18 FA (secreted, % of intake), an estimate for transfer efficiency, decreased with increasing dietary content. This relationship followed a nonlinear decay function. When the dietary content of these FA exceeded a certain threshold (about 0.2, 0.8, and 2.8% of DM for ALA, LLA, and total C18 FA, respectively) the recovery in milk remained constant at about 5, 10, and 82% of the ingested ALA, LLA, and total C18 FA, respectively. At dietary proportions below 0.01% ALA and 1.5% total C18 FA of DM, their apparent recovery in milk fat exceeded 100%. In conclusion, a general inverse relationship between dietary C18 FA and the corresponding apparent recovery in milk fat seems to exist. Within this frame, the effect of different types of feeding regimens on the eventual milk C18 FA profile varies. Among them, grazing pasture appears to provide the most variable properties.

Substitution of common concentrates with by-products modulated ruminal fermentation, nutrient degradation, and microbial community composition in vitro.

A rumen simulation technique was used to evaluate the effects of the complete substitution of a common concentrate mixture (CON) with a mixture consisting solely of by-products from the food industry (BP) at 2 different forage-to-concentrate ratios on ruminal fermentation profile, nutrient degradation, and abundance of rumen microbiota. The experiment was a 2×2 factorial arrangement with 2 concentrate types (CON and BP) and 2 concentrate levels (25 and 50% of diet dry matter). The experiment consisted of 2 experimental runs with 12 fermentation vessels each (n=6 per treatment). Each run lasted for 10d, with data collection on the last 5d. The BP diets had lower starch, but higher neutral detergent fiber (NDF) and fat contents compared with CON. Degradation of crude protein was decreased, but NDF and nonfiber carbohydrate degradation were higher for the BP diets. At the 50% concentrate level, organic matter degradation tended to be lower for BP and CH4 formation per unit of NDF degraded was also lower for BP. The BP mixture led to a higher concentration of propionate and a lower acetate-to-propionate ratio, whereas concentrations of butyrate and caproate decreased. Concentrate type did not affect microbial community composition, except that the abundance of bacteria of the genus Prevotella was higher for BP. Increasing the concentrate level resulted in higher degradation of organic matter and crude protein. At the higher concentrate level, total short-chain fatty acid formation increased and concentrations of isobutyrate and valerate decreased. In addition, at the 50% concentrate level, numbers of protozoa increased, whereas numbers of methanogens, anaerobic fungi, and fibrolytic bacteria decreased. No interaction was noted between the 2 dietary factors on most variables, except that at the higher concentrate level the effects of BP on CH4 and CO2 formation per unit of NDF degraded, crude protein degradation, and the abundance of Prevotella were more prominent. In conclusion, the results of this study suggest that BP in the diet can adequately substitute CON with regard to ruminal fermentation profile and microbiota, showing even favorable fermentation patterns when fed at 50% inclusion rate.

Fortification of dried distillers grains plus solubles with grape seed meal in the diet modulates methane mitigation and rumen microbiota in Rusitec.

The role of dried distillers grains plus solubles (DDGS) and associative effects of different levels of grape seed meal (GSM) fortified in DDGS, used as both protein and energy sources in the diet, on ruminal fermentation and microbiota were investigated using rumen-simulation technique. All diets consisted of hay and concentrate mixture with a ratio of 48:52 [dry matter (DM) basis], but were different in the concentrate composition. The control diet contained soybean meal (13.5% of diet DM) and barley grain (37%), whereas DDGS treatments, unfortified DDGS (19.5% of diet DM), or DDGS fortified with GSM, either at 1, 5, 10, or 20% were used entirely in place of soybean meal and part of barley grain at a 19.5 to 25% inclusion level. All diets had similar DM, organic matter, and crude protein contents, but consisted of increasing neutral detergent fiber and decreasing nonfiber carbohydrates levels with DDGS-GSM inclusion. Compared with the soy-based control diet, the unfortified DDGS treatment elevated ammonia concentration (19.1%) of rumen fluid associated with greater crude protein degradation (~19.5%). Methane formation decreased with increasing GSM fortification levels (≥ 5%) in DDGS by which the methane concentration significantly decreased by 18.9 to 23.4 and 12.8 to 17.6% compared with control and unfortified DDGS, respectively. Compared with control, unfortified DDGS decreased butyrate proportion, and GSM fortification in the diet further decreased this variable. The proportions of genus Prevotella and Clostridium cluster XIVa were enhanced by the presence of DDGS without any associative effect of GSM fortification. The abundance of methanogenic archaea was similar, but their composition differed among treatments; whereas Methanosphaera spp. remained unchanged, proportion of Methanobrevibacter spp. decreased in DDGS-based diets, being the lowest with 20% GSM inclusion. The abundance of Ruminococcus flavefaciens, anaerobic fungi, and protozoa were decreased by the GSM inclusion. As revealed by principal component analysis, these variables were the microorganisms associated with the methane formation. Grape seed meal fortification level in the diet decreased DM and organic matter degradation, but this effect was more related to a depression of nonfiber carbohydrates degradation. It can be concluded that DDGS fortified with GSM can favorably modulate ruminal fermentation.

Meta-analysis of the effects of essential oils and their bioactive compounds on rumen fermentation characteristics and feed efficiency in ruminants.

The present study aimed at investigating the effects of essential oils and their bioactive compounds (EOBC) on rumen fermentation in vivo as well as animal performance and feed efficiency in different ruminant species, using a meta-analysis approach. Ruminant species were classified into 3 classes consisting of beef cattle, dairy cattle, and small ruminants. Two datasets (i.e., rumen fermentation and animal performance) were constructed, according to the available dependent variables within each animal class, from 28 publications (34 experiments) comprising a total of 97 dietary treatments. In addition, changes in rumen fermentation parameters relative to controls (i.e., no EOBC supplementation) of all animal classes were computed. Data were statistically analyzed within each animal class to evaluate the EOBC dose effect, taking into account variations of other variables across experiments (e.g., diet, feeding duration). The dose effect of EOBC on relative changes in fermentation parameters were analyzed across all animal classes. The primary results were that EOBC at doses <0.75 g/kg diet DM acted as a potential methane inhibitor in the rumen as a result of decreased acetate to propionate ratio. These responses were more pronounced in beef cattle (methane, P = 0.001; acetate to propionate ratio, P = 0.005) than in small ruminants (methane, P = 0.068; acetate to propionate ratio, P = 0.056) and in dairy cattle (P > 0.05), respectively. The analysis of relative changes in rumen fermentation variables suggests that EOBC affected protozoa numbers (P < 0.001) but only high doses (>0.20 g/kg DM) of EOBC had an inhibitory effect on this variable whereas lower doses promoted the number. For performance data, because numbers of observations in beef cattle and small ruminants were small, only those of dairy cattle (DMI, milk yield and milk composition, and feed efficiency) were analyzed. The results revealed no effect of EOBC dose on most parameters, except increased milk protein percentage (P< 0.001) and content (P = 0.006). It appears that EOBC supplementation can enhance rumen fermentation in such a way (i.e., decreased acetate to propionate ratio) that may favor beef production. High doses of EOBC do not necessarily modify rumen fermentation or improve animal performance and feed efficiency. Furthermore, additional attention should be paid to diet composition and supplementation period when evaluating the effects of EOBC in ruminants.

Effects of species-diverse high-alpine forage on in vitro ruminal fermentation when used as donor cow's feed or directly incubated.

Alpine forages are assumed to have specific effects on ruminal digestion when fed to cattle. These effects were investigated in an experiment from two perspectives, either by using such forages as a substrate for incubation or as feed for a rumen fluid donor cow. In total, six 24-h in vitro batch culture runs were performed. Rumen fluid was collected from a non-lactating donor cow after having grazed pastures at ∼2000 m above sea level for 2, 6 and 10 weeks. These 'alpine runs' were compared with three lowland samplings from before and 2 and 6 weeks after the alpine grazing where a silage-concentrate mix was fed. In each run, nine replicates of four forages each were incubated. These forages differed in type and origin (alpine hay, lowland ryegrass hay, grass-maize silage mix, pure hemicellulose) as well as in the content of nutrients. Concentrations of phenolic compounds in the incubated forages were (g/kg dry matter (DM)): 20 (tannin proportion: 0.47), 8 (0.27), 15 (0.52) and 0 (0), respectively. Crude protein was highest in the silage mix and lowest with hemicellulose, whereas the opposite was the case for fiber. The total phenol contents (g/kg DM) for the high altitude and the lowland diet of the donor cow were 27 (tannins: 0.50 of phenols) and 12 (0.27), respectively. Independent of the origin of the rumen fluid, the incubation with alpine hay decreased (P < 0.05) bacterial counts, fermentation gas amount, volatile fatty acid (VFA) production as well as ammonia and methane concentrations in fermentation gas (the latter two being not lower when compared with hemicellulose). Alpine grazing of the cow in turn increased (P < 0.001) bacterial counts and, to a lesser extent, acetate proportion compared with lowland feeding. Further, alpine grazing decreased protozoal count (P < 0.05) and VFA production (P < 0.001) to a small extent, whereas methane remained widely unchanged. There were interactions (P < 0.05) between forage type incubated and feeding period of the donor cow in protozoal counts, acetate:propionate ratio, fermentation gas production and its content of methane, in vitro organic matter digestibility and metabolizable energy. Although increased phenolic compounds were the most consistent common property of the applied alpine forages, a clear attribution to certain effects was not possible in this study. As a further result, adaptation (long-term for donor cow, short term for 24 h incubations) appears to influence the expression of alpine forage effects in ruminal fermentation.

Evidence for the inhibition of the terminal step of ruminal alpha-linolenic acid biohydrogenation by condensed tannins.

Effects of condensed tannins (CT), either via extract or plant-bound, and saponin extract on ruminal biohydrogenation of alpha-linolenic acid (ALA) were investigated in vitro. Grass-clover hay served as basal diet (control). The control hay was supplemented with extracts contributing either CT from Acacia mearnsii [7.9% of dietary dry matter (DM)] or saponins from Yucca schidigera (1.1% of DM). The fourth treatment consisted of dried sainfoin (Onobrychis viciifolia), a CT-containing forage legume, in an amount also providing 7.9% CT in dietary DM. All diets were supplemented with linseed oil at a level contributing 60% of total dietary ALA in all treatments. Diets were incubated for 10 d (n = 4) in the rumen simulation technique system, using the last 5 d for statistical evaluation. Fatty acids were analyzed in feed, feed residues, incubation fluid, and its effluent. Data were subjected to ANOVA considering diet and experimental run as main effects. Both CT treatments reduced ruminal fiber and crude protein degradation, and lowered incubation fluid ammonia concentration. Only the CT extract suppressed methane formation and shifted microbial populations toward bacteria at cost of protozoa. The saponin extract remained without clear effects on fermentation characteristics except for increased protozoal counts. The extent of ALA biohydrogenation was 20% less with the CT plant, but this probably resulted from reduced organic matter degradability rather than from an inhibition of biohydrogenation. After incubation analysis of incubation fluid effluent and feed residues showed a considerable proportion of the 3 biohydrogenation intermediates, cis-9, trans-11, cis-15 C18:3, trans-11, cis-15 C18:2, and trans-11 C18:1, which did not occur in the initial feeds. Only the CT-extract diet led to a different profile in the effluent compared with the control diet with trans-11 C18:1 being considerably increased at cost of C18:0. This could have been achieved by suppressing protozoa and enhancing the bacterial population, thus removing potential microbes involved in biohydrogenation and increasing competition between bacteria involved in biohydrogenation and others. The elevation of trans-11 C18:1 as the precursor of cis-9, trans-11 conjugated linoleic acid formed in body tissue and mammary gland is probably favorable from a human health point of view.

Early deposition of n-3 fatty acids from tuna oil in lean and adipose tissue of fattening pigs is mainly permanent.

A total of 600 crossbred pigs, whereof 56 were randomly selected for more in-depth studies of carcass and pork quality, were employed to test different tuna oil feeding regimens. The focus was put on the efficiency to enrich lean and adipose tissue with n-3 fatty acids and the expression of adverse side-effects on performance, carcass, and pork quality. The 4 treatments were 0% tuna oil in diet (T0; control), 1% of unrefined tuna oil in diet fed from 35 to 90 kg of BW (T1), and 3% of unrefined tuna oil in diet offered during the early (35 to 60 kg of BW; T3-E) or late stage of fattening (75 to 90 kg of BW; T3-L). With this arrangement, pigs consumed equal lifetime amounts of tuna oil (approximately 1.6 kg per pig). None of the tuna oil treatments had significant effects on performance. There were no differences in carcass quality among tuna oil groups except for group T3-E where carcasses and loin chops were fatter than those of the other groups. Water-holding capacity and texture of the loin as well as firmness and melting properties of the backfat remained widely unaffected by the treatments. Tuna oil feeding resulted in a lighter, less red and less yellow backfat and was found to increase the proportion of n-3 fatty acids to total fatty acids in all treatments. This especially concerned eicosapentaenoic acid and docosahexaenoic acid, but not alpha-linolenic acid. There was also a slight increase in oleic acid, whereas n-6 fatty acids largely decreased. Feeding tuna oil during a short period at the end of fattening (T3-L) or permanently during fattening (T1) proved to be similarly efficient in increasing n-3 fatty acid content of lean and adipose tissue (to about 1.6-fold of T0). By contrast, only two-thirds of this increase was found when the same amount of tuna oil had been fed exclusively during early fattening (T3-E). The decreased efficiency in T3-E was associated with better sensory flavor, overall acceptability grading, and oxidative status. The results show that, particularly under the condition of a continuous supply, much of the n-3 fatty acids ingested in early fattening can be recovered in pork. These findings give farmers flexibility as to when and how pork can be enriched in n-3 fatty acids with fish oil.