The A1/A2 ß-casein genotype of cows, but not their horn status, influences peptide generation during simulated digestion of milk.

The effect of the horn status of cows on their milk composition and quality is a controversial research topic. In this study, 128 milk samples from 64 horned and 64 disbudded Brown Swiss and Original Braunvieh cows were collected from alpine farms where both horned and disbudded cows were grazing on mountain pastures. The samples were analyzed for their detailed composition and protein digestion in a simulated in vitro digestion (INFOGEST). To exclude probable influences on digestion, the ß-casein genotype with its variants A1 and A2 was also included in the study. The effects of horn status and ß-casein genotype were investigated in linear mixed models, which included additional influencing random factors such as Original Braunvieh blood proportion, stage of lactation, and farm. Horn status did not have any effect on milk composition or digestion. In contrast, milk from A1A1 cows showed a different protein digestion than milk of A1A2 and A2A2 cows in the gastric phase, including smaller amounts of ß-casomorphin(BCM)21-associated peptides and larger amounts of BCM11-associated peptides. Abundances of BCM7 did not differ between ß-casein genotypes. At the end of the intestinal phase, the digested milk of A1A1 and A2A2 b-casein genotypes did not differ.

Changes in in vitro ruminal and post-ruminal degradation of tropical tannin-rich legumes due to varying levels of polyethylene glycol.

We evaluated the effects of tannins from Flemingia macrophylla (CIAT 17403) and Calliandra calothyrsus (San Ramón CIAT 22310 and Patulul CIAT 22316) on in vitro ruminal and post-ruminal dry matter and apparent protein degradation. For each tannin source (legumes), different dosages of polyethylene glycol (PEG) (8000 Da) in McDougall buffer were added to achieve ratios of 0:3, 1:3, 2:3 and 3:3 PEG:condensed tannin (CT). Ruminal fluid mixed with McDougall buffer (1:4) was added to tubes containing only legume foliage (control) or PEG-treated legume foliage. For both Calliandra varieties, a higher ruminal dry matter degradation was observed at a PEG:CT ratio of 3:3. For F. macrophylla, no differences were found between 2:3 and 3:3 ratios (p > 0.05), indicating that a PEG:CT ratio of 2:3 might be enough to bind tannins. Increasing PEG:CT ratios increased apparent ruminal degraded protein and ammonia concentration (p < 0.0001) differing among species (species × ratio: p < 0.0001). The degradation of bypass crude protein (dBCP) was influenced by both legume type and PEG:CT ratio (p < 0.0001). For Patulul, as PEG:CT ratio increased, dBCP increased, but after tannin ratio of 2:3, there was not a significant increase, and for San Ramón, dBCP degradation was higher as PEG:CT ratio increased up to 2:3. For Flemingia, dBCP was higher than PEG:CT ratio of 0:3 but not different among 1:3, 2:3 or 3:3. Low concentration of CT (116 mg/g DM) increased the proportion of protein digested in the abomasum, but higher levels of CT (252 mg/g) clearly reduced the proportion of digested CP. For Flemingia, PEG:CT ratio of 2:3 is enough to inactivate tannins, while PEG:CT ratio of 3:3 was needed for Calliandra and consequently increased ruminal degradation of dry mater (rdDM), and crude protein (rdCP), total degradation of dry matter (tdDM), crude protein (tdCP) and ammonia levels.

Modification of milk fatty acid composition by feeding forages and agro-industrial byproducts from dry areas to Awassi sheep.

The study tested the hypothesis that certain underused forages and agro-industrial byproducts available in dry areas may positively influence fatty acid (FA) composition and antioxidative properties of milk by their contents of residual oil or phenolic compounds or both. Sixty multiparous fat-tailed Awassi ewes were allocated to 6 groups in a completely randomized block design. During 50 d, the ewes were group-fed 2.5 kg of dry matter/d per ewe 1 of 6 isonitrogenous and isoenergetic diets (forage:concentrate, 0.3:0.7). The test feeds, comprising 30% of the diets, replaced either barley straw [lentil straw, olive leaves, and Atriplex (saltbush) leaves, rich in phenolic compounds or electrolytes] or conventional concentrate ingredients (olive cake and tomato pomace; ∼10% lipids) from the control diet. The diets containing olive cake and tomato pomace were rich in oleic acid (18:1 cis-9; 27% of total dietary FA) and linoleic acid (18:2 cis-9,cis-12; 37%), respectively. Profiles of FA were determined in individual milk samples drawn on d 0 and in wk 1, 3, 5 and 7. Data was analyzed by repeated measurement analysis. No consistent treatment effects on yield and gross nutrient composition of the milk were observed, although some differences occurred. Milk resulting from the Atriplex leaf diet expressed the highest antiradical activity, which was low with control and olive leaves. Feeding the tomato pomace and olive cake diets decreased the proportions of short- and medium-chain FA, whereas oleic acid clearly increased in proportion to total FA. Olive leaves most effectively increased rumenic acid (18:2 cis-9,trans-11) and α-linolenic acid (18:3 cis-9,cis-12,cis-15) in milk fat. This also resulted in the highest α-linolenic acid transfer rate from feed to milk and suggests that olive leaves affect ruminal biohydrogenation at several steps. Several alternative feeds exist with an added value, as they enhance FA with potential health benefits and the stability of the milk with higher antioxidative activity, even though responses to test feeds differed largely. It remains to be investigated whether combinations of these feeds would be complementary in these beneficial effects.

In vitro evidence for the importance of cultivation conditions on the effects of Calliandra tannins on ruminal escape of soybean protein and its post-ruminal degradability.

Purified condensed tannins (CT) extracted from the legume Calliandra calothyrsus (var. San Ramón CIAT 22310), harvested in the dry and the rainy season and cultivated with low or high level of fertilization were added to soybean meal in a ratio of 600 mg/g of the incubated crude protein (CP). Effects on degradation either in ruminal fluid only, or in ruminal fluid followed by incubation in HCl/pepsin, were evaluated using a modified two-step in vitro method. Season was found to have larger effects on in vitro ruminal and post-ruminal CP degradation than fertilization. Condensed tannins from the rainy season harvest reduced ruminal CP degradation less than that from the dry season harvest. They had also less negative effects on the degradability of rumen escape protein and enhanced the proportion of post-ruminally degraded CP more than CT from the dry season harvest. An increase in level of fertilization reduced ruminal CP degradation in CT from the rainy season plants but this was not associated with effects on post-ruminal degradation. The study demonstrated the importance of environmental factors for the efficiency of CT in modifying ruminal and post-ruminal CP degradation.

The ACTH challenge and its repeatability in fattening bulls-influences of physiological state, challenge time standardization, and horn status.

Adrenocorticotropic hormone (ACTH) challenges are frequently performed in repetition when evaluating stress or welfare in animals. To our knowledge, the repeatability of ACTH challenges in cattle, although fundamental to further studies of this type, has not yet been the subject of research. Therefore, the objective of this study was to evaluate the repeatability of ACTH challenges in fattening bulls of different horn status. Eight-one bulls were subjected to 3 consecutive ACTH challenges. The first challenge (C1) was performed in calves aged 1.5 mo. Subsequently, animals were characterized as high or low cortisol responders and either disbudded or left with horns. They were then assigned to 1 of 3 rearing groups: a horned group (H+), a disbudded group (H-), and a mixed group (M; 50% horned and 50% disbudded), with each group containing an equal number of high and low responders. The second ACTH challenge (C2) was performed at the age of 11 mo. Time of day (TOD) of challenge was either fixed (ST = same TOD) or alternated (AT = alternate TOD) between C1 and C2. The third ACTH challenge (C3) was performed 7 d after and at the same TOD as C2. Saliva samples were taken 60 and 30 min before and 30, 60, 90, 120, and 150 min after each intravenous ACTH injection. The area under the curve (AUC) was calculated with respect to both ground (AUCG) and to increase (AUCI). The AUCI increased markedly between C1 and C2 (P < 0.05) in ST bulls, and no effects were observed for AUCG between C1 and C2 in ST or AT bulls, nor for any AUC between C2 and C3 (P > 0.1). The overall repeatability of AUCG and AUCI between C1 and C2, reflecting the repeatability between 2 different physiological states, was poor and moderate, respectively, for ST bulls (AUCG: r = 0.24, P > 0.1, intraclass correlation coefficient (ICC) = 0.21; AUCI: r = 0.48, P < 0.01, ICC = 0.41) and lacked in AT bulls (AUCG: r = 0.07, P > 0.1; ICC = 0.03; AUCI: r = 0.08, P > 0.1, ICC = 0.06). The repeatability of AUCG and AUCI between C2 and C3, reflecting the repeatability within the same physiological state, was moderate (AUCG: r = 0.59, P < 0.001; ICC = 0.53; AUCI: r = 0.58, P < 0.001, ICC = 0.52). Assignment to high and low responder groups based on peak cortisol concentration in C1 did not persist over time. H+ bulls showed higher AUCI in C2 and C3 (P < 0.1 and P < 0.05, respectively) than H- bulls. The M group differed from the H- group only in C3 (P < 0.05). Thus, the effect of horn status on ACTH challenges needs further investigation. In conclusion, our results report poor repeatability of the cortisol response to ACTH challenges for challenges performed in different physiological states and moderate repeatability for challenges performed within the same physiological state. Moreover, they point out the importance of standardization of TOD when performing repeated ACTH challenge.

Effect of different potassium levels in hay on acid-base status and mineral balance in periparturient dairy cows.

Forages commonly used in dry cow rations contain high K concentrations. This results in a high dietary cation-anion difference (DCAD), which can compromise the calcium homeostasis of periparturient cows. The aim of this study was to determine the effect of 2 types of hay, fed during the prepartum period and differing in their K concentrations, on the peripartum acid-base status and mineral balance of dairy cows. During the prepartum period, the cows of group K(33) (n = 6) received a diet based on hay with a high K concentration (33 g/kg of DM), whereas the cows of group K(13) (n = 6) received a diet based on hay with a low K concentration (13 g/kg of DM). Both experimental diets were formulated to be isoenergetic and isonitrogenous. After calving, all cows received the same diet based on hay K(33). Blood and urine samples were taken on d 14, 7, and 3 before parturition, at parturition, and then daily during the first 8 d after calving. Concentrations of minerals were analyzed in both blood and urine. Creatinine was also measured in urine for the calculation of the mineral:creatinine ratio. The acid-base parameters in blood (pH and HCO(3)(-) concentration) and urine (pH, net acid-base excretion, and base-acid quotient) were determined on d 14, 7, and 3 before parturition, at parturition, and on d 1 after parturition. The use of hay K(13) reduced the DCAD value of the prepartum diet by half (195 vs. 514 mEq/kg of DM). No significant differences between the 2 groups were observed for blood acid-base indicators or plasma minerals except for the Mg plasma concentration, which tended to be higher in group K(13) from d 3 prepartum to d 2 after calving. In group K(13), urinary Ca excretion tended to be higher from d 3 prepartum to d 1 after parturition than that in group K(33). On d 3 before parturition, urinary pH and net acid-base excretion were significantly lower in group K(13) than in group K(33). On d 14, 7, and 3 before parturition, base-acid quotient was significantly lower in group K(13) than in group K(33). In group K(13), daily feed intake and hence daily intake of Ca, P, and Mg during d 3 and 4 after parturition were higher than in group K(33). The decrease of the DCAD in positive ranges by feeding a low-K hay before parturition induced a reduction of the metabolic alkalotic charge, as observed in acid-base parameters in urine, and increased the availability of Ca and P as a result of higher feed intake at the onset of lactation.

Pre-slaughter stress and horn status influence physiology and meat quality of young bulls.

The study used 81 young bulls, half of which were disbudded at 7 weeks of age. The effects of horn status during rearing and of acute physical and emotional stress just before slaughter on the physiological status at slaughter and subsequent meat quality were investigated. Bulls were reared in groups containing i) only bulls with horns, ii) only bulls without horns, or iii) mixed (half with, and half without horns). Bulls of each rearing condition were assigned to one of two slaughter conditions: with limited (LS) or with supplementary stress (SS). LS resulted in lower heart rates, stress hormone concentrations and carcass temperature, whereas SS resulted in faster post mortem pH decline and lower juiciness. Horned bulls from unmixed rearing groups had lower early pm temperature, shorter sarcomeres, and lower tenderness compared to disbudded bulls. Correlations and regression analysis revealed relationships between physiological indicators, mainly heart rate before slaughter, and meat quality, including water-holding capacity and indicators of proteolysis.

Effects of Sapindus saponaria fruits on ruminal fermentation and duodenal nitrogen flow of sheep fed a tropical grass diet with and without legume.

Six adult African-type hair sheep (BW = 40.3 +/- 6.3 kg) fitted with ruminal and duodenal cannulas were subjected to four treatments. Sheep were offered basal diets at a rate of 80 g of DM/kg of metabolic BW (equivalent to ad libitum access) consisting either of a low-quality grass hay (Brachiaria dictyoneura, 3.7% CP, DM basis) alone or in combination with a forage legume (Cratylia argentea, 18.6% CP, DM basis) in a 3:1 ratio (DM basis). In addition, 0 or 8 g of DM of Sapindus saponaria fruits (12.0% crude saponins, DM basis) per kilogram of metabolic BW was administered intraruminally. Supplementation of C. argentea increased intakes of OM (+21%; P < 0.01) and CP (+130%; P < 0.001), as well as ruminal fluid ammonia N concentrations (from 2.40 to 8.43 mg/dL; P < 0.001). Apparent OM and N digestibilities were not affected by legume addition, but ADF digestibility decreased by 10% (P < 0.01). Total ruminal VFA concentration was unchanged, but acetate:propionate was lower (P < 0.01) and isobutyrate proportion was greater (P < 0.001) with the legume addition. Legume supplementation increased duodenal flows of total N (+56%; P < 0.001), nonammonia N (+52%; P < 0.001), ruminal escape N (+80%; P < 0.001), and microbial N (+28%; P < 0.05). Microbial efficiency was not affected by legume addition. Supplementation of S. saponaria increased (P < 0.05) dietary OM intake by 14%, but had no effect on CP intake and ruminal fluid ammonia concentration or on OM and N digestion. Digestibility of ADF was decreased (P < 0.01) by 10% with S. saponaria as was acetate:propionate (P < 0.001) and the isobutyrate proportion (P < 0.001). Ruminal protozoa counts increased (P < 0.01) by 67% with S. saponaria. Duodenal N flows were not significantly affected by S. saponaria supplementation, except for microbial N flow (+34%; P < 0.01). Microbial efficiency was greater (P < 0.05) by 63% with the addition of S. saponaria. Few interactions between legume and S. saponaria supplementation were observed. The NDF digestibility was decreased with S. saponaria in the grass-alone diet, but not in the legume-supplemented diet (interaction; P < 0.05). Interactions were absent in ruminal fermentation measures and duodenal N flow, indicating that effects were additive. Results suggest that, even when not decreasing ruminal protozoa count, supplementation of S. saponaria fruits is a beneficial way to improve ruminal VFA profile, microbial efficiency, and duodenal flow of microbial protein in sheep fed tropical grass-alone or grass-legume diets.

Investigation of sainfoin (Onobrychis viciifolia) cultivar differences on nitrogen balance and fecal egg count in artificially infected lambs.

Research in ruminant nutrition and helminth control with forages, which contain condensed tannins (CT), suggests that varying responses may depend not only on CT concentration but also on CT composition. An experiment was designed to test this by feeding 2 dried sainfoin cultivars (Visnovsky and Perly), which differed in CT properties, to lambs that were artificially infected with the abomasal blood-sucking nematode Haemonchus contortus. Twenty-four infected lambs received 1 of these 2 cultivars; the feeds were either untreated or treated with the CT-binding polyethylene glycol over 4 wk (n = 6). The 2 cultivars were also fed to 2 × 6 uninfected lambs. Nutrient digestibility, N balance, ADG, plasma urea, together with indicators of infection [fecal egg count (FEC), abomasal worm count, per capita female fecundity, erythrocytic indices, and serum protein], were determined. The specific effects of sainfoin cultivar, CT, and infection were evaluated by contrast analysis. Digestibility of both NDF and ADF were less (P < 0.001) with Perly compared with Visnovsky. The apparent nutrient digestibility was reduced (P < 0.001) by CT. However, no clear cultivar effects were evident on N excretion and retention. Condensed tannins reduced (P = 0.05) body N retention and shifted (P < 0.001) N excretion from urine to feces. Unlike cultivar and CT, infection decreased (P = 0.002) ADG. Plasma urea concentration was decreased (P = 0.007) in Perly- compared with Visnovsky-fed lambs and was decreased (P < 0.001) by CT. Plasma concentrations of essential and semiessential AA were increased (P < 0.001) by CT. The groups of infected lambs did not clearly differ in abomasal worm counts and erythrocytic indicators. In the last 2 to 3 wk of the experiment, FEC was decreased (P ≤ 0.01) when feeding CT. The lack of substantial cultivar effects suggests that the differences in CT properties may have been too small to result in nutritional and anthelmintic effects. The present results indicate that sainfoin CT had a mitigating effect on FEC and, consequently, pasture infectivity. However, the reduction was too small to expect any significant benefits in an Haemonchus-dominated system. Therefore, the use of sainfoin for controlling H. contortus should only be one component within an integrated worm control system.

Effect of the tropical tannin-rich shrub legumes Calliandra calothyrsus and Flemingia macrophylla on methane emission and nitrogen and energy balance in growing lambs.

The objective of this study was to test whether the use of tannin-rich shrub legume forage is advantageous for methane mitigation and metabolic protein supply at unchanged energy supply when supplemented in combination with tannin-free legumes to sheep. In a 6 × 6 Latin-square design, foliage of two tannin-rich shrub legume species (Calliandra calothyrsus and Flemingia macrophylla) were used to replace either 1/3 or 2/3, respectively, of a herbaceous high-quality legume (Vigna unguiculata) in a diet composed of the tropical grass Brachiaria brizantha and Vigna in a ratio of 0.55 : 0.45. A Brachiaria-only diet served as the negative control. Each experimental period lasted for 28 days, with week 3 serving for balance measurement and data collection inclusive of a 2-day stay of the sheep in open-circuit respiration chambers for measurement of gaseous exchange. While Vigna supplementation improved protein and energy utilisation, the response to the partial replacement with tannin-rich legumes was less clear. The apparent total tract digestibilities of organic matter, NDF and ADF were reduced when the tannin-rich plants partially replaced Vigna, and the dose-response relationships were mainly linear. The tannin-rich plants caused the expected redistribution of more faecal N in relation to urinary N. While Flemingia addition still led to a net body N retention, even when fed at the higher proportion, adding higher amounts of Calliandra resulted in body protein mobilisation in the growing lambs. With respect to energy, supplementation of Vigna alone improved utilisation, while this effect was absent when a tannin-rich plant was added. The inclusion of the tannin-rich plants reduced methane emission per day and per unit of feed and energy intake by up to 24% relative to the Vigna-only-supplemented diet, but this seems to have been mostly the result of a reduced organic matter and fibre digestion. In conclusion, Calliandra seems less apt as protein supplement for ruminants while Flemingia could partially replace a high-quality legume in tropical livestock systems. However, methane mitigation would be small due to associated reductions in N and energy retention.

Nitrogen and mineral balance of lambs artificially infected with Haemonchus contortus and fed tanniferous sainfoin (Onobrychis viciifolia).

Tanniferous temperate legumes are assumed to possess anthelmintic properties, but it is unclear whether this is the direct result of condensed tannins (CT) or is mediated indirectly via an improved metabolic protein supply. A metabolism experiment was conducted to differentiate between these factors by feeding the CT plant sainfoin (19.7% CP in DM) to lambs infected with the abomasal blood-sucking nematode Hemonchus contortus. A total of 18 infected lambs were fed sainfoin either untreated or treated with polyethylene glycol, a CT-inactivating agent, or a grass-clover mixture (13.2% CP in DM) over 3 wk (n = 6). Six uninfected lambs received the grass-clover mixture as a control. In addition to indicators of infection (fecal egg count, packed-cell volume, abomasal worm burden, and serum protein), nutrient digestibility, the balance of N and selected minerals, ruminal fluid characteristics, and plasma AA levels were determined mostly in the final experimental week. The specific effects of the sainfoin CT, the extra CP with sainfoin, and the infection were statistically evaluated by contrast analysis. The sainfoin CT exerted no beneficial effects on resilience to nematode infection and exerted only minor effects on ruminal ammonia or blood urea concentrations and the excretory pattern of N. Plasma alanine, aspartate, and proline concentrations tended to be greater (P < or = 0.09) because of the sainfoin CT, whereas the other AA remained unaffected. Intake of the mineral supplement was lower (P < 0.001) for lambs fed sainfoin compared with those fed sainfoin treated with polyethylene glycol. Feeding the high-protein sainfoin instead of the grass-clover mixture increased (P < 0.001) N retention and apparent OM digestibility, whereas digestibility of NDF and ADF were decreased (P < 0.001). Feeding sainfoin also decreased (P < or = 0.04) plasma alanine, glycine, isoleucine, and total nonessential AA compared with the grass-clover mixture. Although fecal egg count, worm burden, and packed cell volume were not affected by the greater CP supply associated with sainfoin feeding, serum albumin level was increased (P = 0.008). The lack of effects of sainfoin on resilience to nematode infection might have been the result of the unexpectedly low CT content (3.6% in DM) of the material used. It cannot be excluded that longer term feeding of this batch of sainfoin might have been effective.

Influence of different calcium contents in diets supplemented with anionic salts on bone metabolism in periparturient dairy cows.

At the initiation of lactation, Ca homeostatic mechanisms have to react to a tremendous increase in demand for Ca. Mobilization of Ca from bone and increased absorption from the gastrointestinal tract are required to re-establish homeostasis. It has been shown that dietary anions play an important role in the prevention of milk fever by mobilizing Ca from bone and by increasing Ca absorption in the GI tract. The purpose of the present study was to investigate the influence of different Ca contents in diets supplemented with anionic salts on bone metabolism of dairy cows. Twenty-four holstein cows (housed inside, second to fourth lactation) without a milk fever history were divided into four groups (A, B, C, D). Each group was fed a different diet which was given from day 263 of gestation till the day of parturition. Group A and B received a low calcium diet (4 g/kg DM) whereas group C and D received a high Ca diet (8 g/kg DM). In addition group B and D received anionic salts. The DCAD was calculated with the formula: DCAD (mEq/kg DM)=(0.2 Ca2++0.16 Mg2++Na++K+)-(Cl-+0.6 S2-+0.65 P3-). Blood and urine samples were collected on days 256, 270 and 277 of gestation, on the day of parturition as well as the following 5 days and on days 9, 14 and 19 after parturition. Serum Ca, P, Mg, ICTP, OC, VITD, PTH and urinary pH were analysed. The bone resorption marker ICTP showed a significant increase after parturition in all the groups. On the contrary, the bone formation marker OC decreased after parturition in all the groups. The VITD concentrations in group D and the urinary pH in group B were significantly lower compared to the other groups (p<0.05). The Ca concentrations tended to be higher in group B around parturition than in all the other groups. No significant influence of the four different diets on all the other parameters could be shown. In conclusion, this data showed that the addition of anions and the different Ca contents had no significant influence on bone resorption and bone formation markers. This may be because of the fact that the dietary cation-anion balance was not low enough (DCAD-group A: 181 mEq/kg DM, group B: -48 mEq/kg DM, group C: 210 mEq/kg DM and group D: 28 mEq/kg DM) to induce a metabolic acidosis with all its positive effects on calcium metabolism.