Effects of feeding rumen-protected linseed fat to postpartum dairy cows on plasma n-3 polyunsaturated fatty acid concentrations and metabolic and reproductive parameters.

High-yielding dairy cows experience a negative energy balance and inflammatory status during the transition period. Fat supplementation increases diet energy density, and plasma n-3 polyunsaturated fatty acids (PUFA) have been proposed to improve immune function. This study tested the hypothesis that dietary supplementation with a rumen-protected and n-3 PUFA-enriched fat could ameliorate both the energetic deficit and immune status of postpartum high-yielding dairy cows, improving overall health and reproductive efficiency. At 11 d in milk (DIM), cows were randomly allocated to groups (1) n-3 PUFA (n = 29), supplemented with encapsulated linseed oil supplying additional up to 64 g/d (mean 25 ± 4 g/d) of α-linolenic acid (ALA), or (2) control (n = 31), supplemented with hydrogenated palm oil without ALA content. Fat supplements of the n-3 PUFA and control groups were available through an automated, off-parlor feeding system, and intake depended on the cow's feeding behavior. Plasma ALA concentrations were higher in n-3 PUFA than control cows, following a linear relation with supplement ingestion, resulting in a lower n-6/n-3 ratio in plasma. Metabolic parameters (body condition score and glucose and ß-hydroxybutyric acid blood concentrations) were unaffected, but milk yield improved with increased intake of fat supplements. Plasma total adiponectin concentrations were negatively correlated with ingestion of n-3 PUFA-enriched fat supplement, following a linear relation with intake. Conception rate to first AI increased with higher intake of both fats, but a decrease of calving-to-conception interval occurred only in n-3 PUFA cows. Postpartum ovarian activity and endometrial inflammatory status at 45 DIM were unaffected. In conclusion, this study evinced a positive linear relation between rumen-protected linseed fat intake and plasma n-3 PUFA concentrations, which modulated adiponectin expression and improved reproductive parameters.

Fatty Acid Content and Composition of the Morphological Fractions of Cistus Ladanifer L. and Its Seasonal Variation.

Cistus ladanifer L. is a shrub from Cistaceae family, widespread in Mediterranean countries. Fatty acids (FA) have multiple roles in plants and are involved in adaption mechanisms to environmental conditions. This work evaluated the FA content and composition of each morphological fraction of C. ladanifer (leaves, stems, flower buds, flowers and seed heads) throughout a full year. Cistus ladanifer plants were collected in southern Portugal, during four consecutive seasons (18 plants/season), and the different morphological plant fractions (leaves, stems, flower buds, flowers and seed heads) were separated. Cistus ladanifer morphological fractions showed distinct FA compositions, being possible to discriminate three groups-the leaves that showed to be dominated by saturated FA (main 20:0) and contain branched-chain FA (iso-19:0 and iso-21:0); the stems that are composed mainly by SFA (main 22:0); and the reproductive organs that showed higher contents of polyunsaturated FA (PUFA) and the 16:0 as the main SFA. The FA composition of leaves changed over seasons, with replacement of the PUFA by monounsaturated FA and branched-chain FA during hot seasons. Regarding the other C. ladanifer morphological fractions, the FA composition was more stable over seasons, suggesting that leaves are more prone to adaptations to environmental changes.

Effect of increasing amounts of olive crude phenolic concentrate in the diet of dairy ewes on rumen liquor and milk fatty acid composition.

Agro-industrial by-products contain several secondary plant metabolites, such as polyphenols, tannins, saponins, and essential oils. The effects of these compounds on animal metabolism may vary significantly according to the dose, the chemical nature of the molecules, and the overall composition of the diet. In the Mediterranean area, the olive oil extraction is associated with 2 by-products: olive pomace and wastewater, both rich in polyphenols. In particular, wastewater may be further processed to obtain olive crude phenolic concentrate (OCPC). An experiment was carried out aiming to evaluate animal performance, milk fatty acid (FA) profile, diversity of rumen microbial population, and rumen liquor FA profile in dairy ewes fed diets containing extruded linseed (EL) and increasing doses of OCPC. Twenty-eight Comisana ewes in mid lactation were allotted to 4 experimental groups. The experiment lasted 5 wk after 3 wk of adaptation. Diets were characterized by lucerne hay administrated ad libitum and by 800 g/ewe and day of 4 experimental concentrates containing 22% of EL on dry matter and increasing dose of OCPC: 0 (L0), 0.6 (L0.6), 0.8 (L0.8), and 1.2 (L1.2) g of OCPC/kg of dry matter. Milk yield was daily recorded and milk composition was analyzed weekly. At the beginning and at the end of the experiment, samples of rumen liquor were collected to analyze FA profile, changes in rumen microbial population, and dimethylacetal (DMA) composition. The inclusion of OCPC did not affect milk yield and gross composition, whereas milk from L0.8 and L1.2 sheep contained higher concentrations of linoleic (+18%) and α-linolenic acid (+24%) and lower concentration of the rumen biohydrogenation intermediates. A similar pattern was observed for rumen liquor FA composition. No differences were found in the diversity of the rumen microbial population. Total amount of DMA did not differ among treatments, whereas significant differences were found in the concentration of individual DMA; in the diet with a higher amount of OCPC, DMA 13:0, 14:0, 15:0, and 18:0 increased, whereas DMA 16:0 decreased. Probably the presence of polyphenols in the diet induced a rearrangement of bacteria membrane phospholipids as a response to the rumen environment stimulus. Overall, the use of OCPC allowed a significant increase in the polyunsaturated FA content of milk, probably due to a perturbation of the rumen biohydrogenation process. Further studies are needed to understand the correlation between diet composition and the pattern of DMA in rumen liquor.

New insights on the metabolism of ricinoleic acid in ruminants.

Dairy goats were fed a total mixed ration with or without the inclusion of castor oil [40 g/kg of dry matter (DM)] to study the metabolism of ricinoleic acid (12-OH,cis-9-18:1). Ten goats, at 39.7 ± 4.0 d in milk, were individually penned and allocated at random to the 2 experimental diets. Goats were manually milked twice a day. Milk fatty acids (FA) were analyzed as methyl esters and hydroxyl groups were derivatized in trimethylsilyl ethers. Apart from ricinoleic acid, 6 FA were only detected in the milk of the castor oil group. Ricinoleic acid composed 0.3% of total FA in milk of the castor oil group, whereas the hydroxy-FA (8-OH-14:0, 10-OH-16:0, and 12-OH-18:0) and oxo-FA (8-oxo-14:0, 10-oxo-16:0, and 12-oxo-18:0) reached 7.5% of total FA in milk. We anticipate that these FA were derived from the metabolism of ricinoleic acid, although it was not clear if they were produced in the rumen or in the tissues. To confirm that, we conducted in vitro batch incubations repeated for 3 consecutive weeks with castor oil (40 g/kg of DM) and strained rumen fluid from 2 fistulated sheep. To examine the products formed over time, incubation tubes were stopped at 0, 6, 12, 24, 48, and 72 h. The results of the in vitro experiment showed that ricinoleic acid was metabolized in the rumen at a slow rate and the main products formed were 12-OH-18:0 and 12-oxo-18:0, by hydrogenation of the cis-9 double bond, followed by oxidation of the hydroxyl group, respectively. Our results suggest that the 12-OH-18:0 and 12-oxo-18:0 escape rumen and are further metabolized through partial ß-oxidation in ruminant tissues. We propose that the 10-OH-16:0 and 8-OH-14:0 found in goat milk of the castor oil group are successive products of the ß-oxidation of 12-OH-18:0, and the 10-oxo-16:0 and 8-oxo-14:0 are successive products of the 12-oxo-18:0 in tissues. Overall, our results indicate that ricinoleic acid is extensively metabolized in the rumen and tissues, producing mainly oxo- and hydroxy-FA that are further excreted in milk.

Mammary gland and milk fatty acid composition of two dairy goat breeds under feed-restriction.

Goat dairy products are an important source of animal protein in the tropics. During the dry season, pasture scarcity leads animals to lose up to 40% of their body weight, a condition known as Seasonal Weight Loss (SWL) that is one of the major constraints in ruminant production. Breeds with high tolerance to SWL are relevant to understand the physiological responses to pasture scarcity so they could be used in programs for animal breeding. In the Canary Islands there are two dairy goat breeds with different levels of tolerance to SWL: the Palmera, susceptible to SWL; and the Majorera, tolerant to SWL. Fat is one of the milk components most affected by environmental and physiological conditions. This study hypothesises that feed-restriction affects Majorera and Palmera breeds differently, leading to different fatty acid profiles in the mammary gland and milk. An interaction between breed and feed-restriction was observed in the mammary gland. Feed-restriction was associated with an increase in oleic acid and a decrease in palmitic acid percentage in the Palmera breed whereas no differences were observed in the Majorera breed. Palmitic and oleic acids together constituted around 60% of the total fatty acids identified, which suggests that Palmera breed is more susceptible to SWL. In milk, feed-restriction affected both breeds similarly. Regarding the interaction of the breed with the treatment, we also observed similar responses in both breeds, but this influence affects only around 2% of the total fatty acids. In general, Majorera breed is more tolerant to feed-restriction.

Influence of betaine and arginine supplementation of reduced protein diets on fatty acid composition and gene expression in the muscle and subcutaneous adipose tissue of cross-bred pigs.

The isolated or combined effects of betaine and arginine supplementation of reduced protein diets (RPD) on fat content, fatty acid composition and mRNA levels of genes controlling lipid metabolism in pig m. longissimus lumborum and subcutaneous adipose tissue (SAT) were assessed. The experiment was performed on forty intact male pigs (Duroc×Large White×Landrace cross-breed) with initial and final live weights of 60 and 93 kg, respectively. Pigs were randomly assigned to one of the following five diets (n 8): 16·0 % of crude protein (control), 13·0 % of crude protein (RPD), RPD supplemented with 0·33 % of betaine, RPD supplemented with 1·5 % of arginine and RPD supplemented with 0·33 % of betaine and 1·5 % of arginine. Data confirmed that RPD increase intramuscular fat (IMF) content and total fat content in SAT. The increased total fat content in SAT was accompanied by higher GLUT type 4, lipoprotein lipase and stearoyl-CoA desaturase mRNA expression levels. In addition, the supplementation of RPD with betaine and/or arginine did not affect either IMF or total fat in SAT. However, dietary betaine supplementation slightly affected fatty acid composition in both muscle and SAT. This effect was associated with an increase of carnitine O-acetyltransferase mRNA levels in SAT but not in muscle, which suggests that betaine might be involved in the differential regulation of some key genes of lipid metabolism in pig muscle and SAT. Although the arginine-supplemented diet decreased the mRNA expression level of PPARG in muscle and SAT, it did not influence fat content or fatty acid composition in any of these pig tissues.

Replacing cereals with dehydrated citrus pulp in a soybean oil supplemented diet increases vaccenic and rumenic acids in ewe milk.

This study evaluates the effect of the replacement of cereals by dried citrus pulp (DCP) in diets supplemented with 5% of soybean oil, on ewe milk yield and composition, including milk fatty acid (FA). Four Serra da Estrela multiparous ewes in the second month of lactation were used in a double 2×2 Latin square design. Ewes were individually penned and milked twice a day with an 8-h interval. Each experimental period included 14 d of diet adaptation followed by 5d of measurements and sampling. The 2 diets included on dry matter basis 450 g/kg of corn silage and 550 g/kg of either a soybean oil-supplemented concentrate meal containing barley and maize (cereal) or dried citrus pulp (DCP; citrus). Feed was offered ad libitum, considering 10% of orts, and intake was measured daily. Milk yield was higher and dry matter intake tended to be higher with the citrus diet. Milk composition and technological properties for cheese production were not affected by treatments, except for lactose, which was lower with the citrus diet. Replacement of cereals by DCP resulted in a 3-percentage-point decrease of both 18:0 and cis-9-18:1 that were mostly compensated by the 4.19- and 1.68-percentage-point increases of trans-11-18:1 and cis-9,trans-11-18:2, respectively. The intake of C18 FA tended to increase with the citrus diet compared with the cereal diet, but the apparent transfer of 18:2n-6 and of 18:3n-3 did not differ between diets. The milk output of C18 FA increased with the citrus compared with the cereal diet, mostly due to the increase of trans-11-18:1 and cis-9,trans-11-18:2 because the daily milk output of 18:0, trans-10-18:1, cis-9-18:1, 18:2n-6 and 18:3n-3 did not differ between diets. Replacing cereals with DCP in an oil-supplemented diet resulted in a selective increase of trans-11-18:1 and cis-9,trans-11-18:2 in milk, with no major effect on other biohydrogenation intermediates.

Combined effects of dietary arginine, leucine and protein levels on fatty acid composition and gene expression in the muscle and subcutaneous adipose tissue of crossbred pigs.

The cumulative effects of dietary arginine, leucine and protein levels on fat content, fatty acid composition and mRNA levels of genes controlling lipid metabolism in pig longissimus lumborum muscle and subcutaneous adipose tissue (SAT) were investigated. The experiment was performed on fifty-four intact male pigs (Duroc × Pietrain × Large White × Landrace crossbred), with a live weight ranging from 59 to 92 kg. The pigs were randomly assigned to one of six experimental treatments (n 9). The treatments followed a 2 × 3 factorial arrangement, with two levels of arginine supplementation (0 v. 1 %) and three levels of a basal diet (normal protein diet, NPD; reduced protein diet, RPD; reduced protein diet to achieve 2 % of leucine, RPDL). The results showed that dietary arginine supplementation did not affect the intramuscular fat (IMF) content and back fat thickness, but increased the total fat in SAT. This effect was associated with an increase in fatty acid synthase (FASN) and stearoyl-CoA desaturase (SCD) mRNA levels in SAT, which suggests that arginine might be involved in the differential regulation of some key lipogenic genes in pig muscle and SAT. The increase in IMF content under the RPD, with or without leucine supplementation, was accompanied by increased FASN and SCD mRNA levels. Arginine supplementation did not influence the percentage of main fatty acids, while the RPD had a significant effect on fatty acid composition in both tissues. Leucine supplementation of RPD did not change IMF, total fat of SAT and back fat thickness, but increased 16 : 0 and 18 : 1cis-9 and decreased 18 : 2n-6 in muscle.

Effect of coconut processing by-product graded feeding on carcass traits and meat quality of lambs.

The inclusion of by-product coconut mesocarp skins (CMS) in diets was evaluated in feedlot lambs. The objective of this study was to evaluate CMS levels effects on carcass traits and meat quality of lambs. Thirty-five male lambs with an initial body weight of 16.9 ± 2.93 kg were distributed in a completely randomized design with five CMS levels in total dry matter (0; 4.8; 9.6; 14.4 and 19.2%) and fed during 71 d until slaughter. High levels of CMS decreased the intake of dry matter and negatively affected the performance of lambs. Fat and protein contents of Longissimus lumborum muscle (P < 0.05) and the saturated fatty acid (FA) decreased (P < 0.001) whereas polyunsaturated FA increased (P < 0.01) with the inclusion of CMS. The ratio t10/t11-18:1 increased with the inclusion of CMS (P < 0.001). The instrumental color descriptors were unaffected by CMS levels. According to the effects on the investigated meat quality traits we recommend up to 4.8% CMS in diets of confined lambs.

Differential effects of reduced protein diets on fatty acid composition and gene expression in muscle and subcutaneous adipose tissue of Alentejana purebred and Large White × Landrace × Pietrain crossbred pigs.

The present study assessed the effect of pig genotype (fatty v. lean) and dietary protein and lysine (Lys) levels (normal v. reduced) on intramuscular fat (IMF) content, subcutaneous adipose tissue (SAT) deposition, fatty acid composition and mRNA levels of genes controlling lipid metabolism. The experiment was conducted on sixty intact male pigs (thirty Alentejana purebred and thirty Large White × Landrace × Pietrain crossbred), from 60 to 93 kg of live weight. Animals were divided into three groups fed with the following diets: control diet equilibrated for Lys (17·5 % crude protein (CP) and 0·7 % Lys), reduced protein diet (RPD) equilibrated for Lys (13·2 % CP and 0·6 % Lys) and RPD not equilibrated for Lys (13·1 % CP and 0·4 % Lys). It was shown that the RPD increased fat deposition in the longissimus lumborum muscle in the lean but not in the fatty pig genotype. It is strongly suggested that the effect of RPD on the longissimus lumborum muscle of crossbred pigs is mediated via Lys restriction. The increase in IMF content under the RPD was accompanied by increased stearoyl-CoA desaturase (SCD) and PPARG mRNA levels. RPD did not alter backfat thickness, but increased the total fatty acid content in both lean and fatty pig genotype. The higher amount of SAT in fatty pigs, when compared with the lean ones, was associated with the higher expression levels of ACACA, CEBPA, FASN and SCD genes. Taken together, the data indicate that the mechanisms regulating fat deposition in pigs are genotype and tissue specific, and are associated with the expression regulation of the key lipogenic genes.

Enrichment of Brain n-3 Docosapentaenoic Acid (DPA) and Retinal n-3 Eicosapentaenoic Acid (EPA) in Lambs Fed Nannochloropsis oceanica Microalga.

Omega-3 polyunsaturated fatty acids (n-3 PUFAs) have special physiological functions in both brain and retinal tissues that are related to the modulation of inflammatory processes and direct effects on neuronal membrane fluidity, impacting mental and visual health. Among them, the long-chain (LC) n-3 PUFAs, as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are of special importance. Scarce data are available about the fatty acid (FA) composition of the ruminant brain in response to dietary intervention. However, we decided to examine the brain and retina FA composition of lambs supplemented with an EPA-rich microalga feed for 21 days, as it is known that despite the extensive biohydrogenation of dietary PUFAs in the rumen, ruminants can selectively accumulate some n-3 LC-PUFAs in their brain and retinal tissues. Twenty-eight male lambs were fed a control diet, or the same diet further supplemented with Nannochloropsis sp. microalga. Their brains and retina were collected for FA characterization. Overall, the brain FA profile remained unchanged, with little alteration in omega-3 docosapentaenoic acid (DPA) enhancement in both the hippocampus and prefrontal cortex. Retinal tissues were particularly responsive to the dietary intervention, with a 4.5-fold enhancement of EPA in the freeze-dried-fed lambs compared with the control lambs. We conclude that retinal tissues are sensitive to short-term n-3 PUFA supplementation in lambs.

Forage based diet as an alternative to a high concentrate diet for finishing young bulls - Effects on growth performance, greenhouse gas emissions and meat quality.

Two groups of 8 individually housed young crossbred-bulls, in the finishing period, were used to test the effect of a Total Mixed Ration diet with high forage content (54% DM), low starch content (14% DM), supplemented with sunflower seeds (10% DM) (HFS) on growth performance, carcass and meat quality, fatty acid profile and carbon footprint, with reference to a conventional concentrate-based (90% DM) (Control) diet. The experiment lasted 64 days before slaughter. During the experiment, feed intake was monitored daily and live weight every 14 days. Individual CH4 emissions were assessed at 16-days intervals, using a GreenFeed for Large Animal unit. Feed intake and feed conversion ratio were higher for HFS diet, but average daily weight gain and feeding costs were similar for the two diets. Dressing percentage was reduced with HFS diet. The HFS increased redness, yellowness and Chroma of subcutaneous fat, but did not compromise commercial value of the carcasses. Meat colour, shear force, or sensory parameters were not affected by diet. The HFS diet allowed a healthier FA profile, due to the higher proportions of 18:3n-3, t11-18:1 and c9,t11-18:2 and the lower proportion of t10-18:1. The HFS diet did not reduce the carbon footprint in the finishing period of young bulls, due to increased digestive CH4 emissions. The results of this experiment showed that the HFS diet can be an alternative to the conventional diets used in finishing young-bulls. Although it may result in a slight reduction in animal performance, it has a strong impact on reducing dependence on inputs from outside the farm.

Effect of slaughter season and muscle type on the fatty acid composition, including conjugated linoleic acid isomers, and nutritional value of intramuscular fat in organic beef.

BACKGROUND: Consumer awareness regarding the intake of beef of organic origin is strongly associated with the beneficial outcomes to human health, the environment and animal welfare. In this paper the effects of slaughter season and muscle type on the fatty acid composition, conjugated linoleic acid (CLA) isomeric profile, total cholesterol, α-tocopherol and ß-carotene contents and nutritional quality of intramuscular fat in organic beef (n = 30) are reported for the first time. RESULTS: Organic beef showed a very low total lipid content, with seasonal changes in the levels of some fatty acids, CLA isomers, n-6/n-3 polyunsaturated fatty acid (PUFA) ratio, total cholesterol and ß-carotene. In addition, differences between longissimus lumborum (relatively red) and semitendinosus (relatively white) muscles were found for many fatty acids, specific CLA contents, many CLA isomers and both PUFA/saturated fatty acid (SFA) and n-6/n-3 ratios. However, in spite of the seasonal and carcass variations, all organic meats analysed had values of beef similar to pasture-fed cattle. CONCLUSION: From a nutritional perspective, organic meat from both slaughter seasons seems to have high CLA contents, PUFA/SFA and n-6/n-3 indices within the recommended values for the human diet. The data indicate that intramuscular fat in organic meat has a high nutritional value throughout the year.

Fatty Acid Composition of M. Biceps Femoris of Edible Dormouse (Glis glis L.).

This study aimed to investigate the fatty acid (FA) composition of edible dormouse m. biceps femoris in both sexes. More than 20 FA were identified in the muscle, with the 18:1cis-9 (oleic acid) being the most abundant in both sexes, comprising more than 50% of total FA in muscle. The most dominated FA were monounsaturated (MUFA), followed by saturated FA (SFA) and polyunsaturated FA (PUFA), reaching 54.8%, 25.43% and 19.8% of total FA, respectively. Sums of PUFA and n-3 PUFA tended (p > 0.05) to be higher in males than in females. There were no significant differences between sexes on the FA composition. Nevertheless, the 18:2n-6 tended to differ between sexes (p = 0.063). Several long-chain PUFA (LC-PUFA) were detected in dormouse muscle, with the 20:4 n-6 (arachidonic acid, AA) and the 22:6 n-3 (docosahexaenoic acid, DHA) being the most abundant in both sexes. The relatively high stearoyl-CoA desaturase (SCD) indexes and the large concentration of 18:1cis-9 in dormouse muscle tissues might point to a low mobilization of the SCD products. Furthermore, finding the unusual FA 20:3 ∆5,∆11,∆14, suggests feeding on leaf and wood lipids of Coniferophytes. We demonstrated sexual size monomorphism in edible dormouse. The literature regarding the composition of dormouse meat is scarce and no studies reported the FA composition of muscle, thus, this work can contribute to increasing the knowledge on edible dormouse physiology and nutritional traits.

Partial replacement of soybean meal with Chlorella vulgaris in broiler diets influences performance and improves breast meat quality and fatty acid composition.

Alternative feed ingredients, such as microalgae, may be more sustainable in comparison to conventional feedstuffs that need large amounts of arable land and are often imported. This study evaluates the effects of Chlorella vulgaris various inclusion levels in the diet of broiler chickens on performance, carcass yield, organ measurements, breast meat quality, fatty acids profile, and antioxidant capacity. A total of two hundred forty 5 d old male Ross 308 broilers were randomly allotted to 4 groups (6 replicates of 10 birds each). Each group received either a control diet or a diet where soybean meal was replaced with 10% (CV10%), 15% (CV15%), or 20% C. vulgaris for 40 d. Performance parameters, carcass and meat traits were evaluated. Compared to the control group, birds supplemented with C. vulgaris (15% and 20%) had lower body weight, weight gain, and feed intake (P < 0.0001), whereas no differences were observed between the control and CV10% groups (P > 0.05). Feed conversion ratio did not differ between control and CV groups. Diets containing C. vulgaris significantly increased ileal digesta viscosity, weight and size of several gastrointestinal compartments, as well as breast muscle yield (P < 0.0001). Incorporation of C. vulgaris resulted in yellower breast muscle (P < 0.0001), with significantly increased chlorophyll a (P < 0.05), chlorophyll b, and total carotenoids contents (P < 0.0001). Inclusion of C. vulgaris decreased bacterial count in meat samples in comparison to controls (P < 0.0001). A 20% C. vulgaris inclusion resulted in higher water holding capacity (P < 0.05) and lower cooking loss (P < 0.05). As dietary C. vulgaris increased, concentrations of DHA + EPA (P < 0.05) and n-3 PUFA (P < 0.0001) increased in breast meat, while the n-6/n-3 PUFA ratio decreased (P < 0.0001). Sensory analysis showed that breast meat from the CV10% group had the highest acceptance score. Overall, dietary concentrations of C. vulgaris of up to 20% improve breast meat quality, whereas 10% of C. vulgaris inclusion is recommended.

Effects of partial substitution of grain by agroindustrial byproducts and sunflower seed supplementation in beef haylage-based finisher diets on growth, in vitro methane production and carcass and meat quality.

Thirty-two bulls were assigned to four total mixed biodiverse haylage-based diets to evaluate the effects of partial substitution of grains by agroindustrial byproducts, sunflower seeds (SS) supplementation and haylage level on growth, in vitro methane production and carcass and meat quality. Dietary treatments included a grain-based diet with 30% grain and haylage:concentrate ratio (H:C) of 60:40 (DM basis) (MCe); a by-product-based diet where 50% of the grain was substituted for by-products (H:C, 60:40) (MBp); a byproducts diet with 10% sunflower seed and 90% (DM) MBp (H:C, 54:46) (MBpSS); and a byproducts, SS diet with increased haylage (H:C, 67.5:32.5) (HBpSS). Dry matter intake and growth rate were lower in HBpSS, but feed conversion ratio was unaffected by diet. In vitro methane emissions were reduced by SS. Meat colour and shear force were similar among diets. Lipid oxidation in cooked meat was reduced and fatty acid composition was improved with SS. Biodiverse haylage-based diets may be a viable option for finishing bulls.

Acetonitrile covalent adduct chemical ionization tandem mass spectrometry of non-methylene-interrupted pentaene fatty acid methyl esters.

Acetonitrile covalent adduct chemical ionization tandem mass spectrometry (CACIMS/MS) has shown to be an efficient method for the identification of double-bond position in homoallylic, conjugated and several polyene non-methylene-interrupted (NMI) fatty acid methyl esters. However, it has not been thoroughly evaluated for NMI highly unsaturated fatty acids (HUFA) with more than four double bonds. Docosahexaenoic acid (DHA)-rich single cell oil (DHASCO(®); Martek Biosciences, Corp.) was partially hydrogenated (partially hydrogenated DHASCO; PHDO) producing ten novel 22:5 and 22:6 HUFA isomers. In single-stage MS, the ratio of [M+54](+)/[M+54-32](+) for the 22:5 and 22:6 isomers indicated the presence of homoallylic or partially conjugated double-bond systems. The CACIMS/MS spectra revealed six 22:5 isomers with diagnostic ions corresponding to the homoallylic 22:5n-6 and 22:5n-3 isomers, and four distinct NMI 22:5 isomers. Diagnostic ions for four 22:6 isomers were identical to the native DHA illustrating that CACIMS/MS is sensitive to double-bond position but not geometry. Three gas chromatography (GC) peaks for partially conjugated 22:6 isomers were also detected and clearly distinguishable from homoallylic 22:6 isomers, but their CACIMS/MS spectra did not yield prominent ions indicative of double-bond position, possibly due to co-elution. Overall, CACIMS/MS was effective for determining double-bond position in NMI 22:5 isomers. Further investigations are warranted to evaluate and determine fragmentation patterns for partially conjugated and NMI 22:6 HUFA.

Condensed Tannins as Antioxidants in Ruminants-Effectiveness and Action Mechanisms to Improve Animal Antioxidant Status and Oxidative Stability of Products.

Condensed tannins (CTs) are widely distributed in plants, and due to their recognized antioxidant activity are considered as possible natural antioxidants for application in ruminant diets. A wide range of CT-rich sources has been tested in ruminant diets, and their effects on animal antioxidant status and oxidative stability of their products are reviewed in the present work. Possible mechanisms underlying the CT antioxidant effects in ruminants are also discussed, and the CT chemical structure is briefly presented. Utilization of CT-rich sources in ruminant feeding can improve the animals' antioxidant status and oxidative stability of their products. However, the results are still inconsistent. Although poorly understood, the evidence suggests that CTs can induce an antioxidant effect in living animals and in their products through direct and indirect mechanisms, which can occur by an integrated and synergic way involving: (i) absorption of CTs with low molecular weight or metabolites, despite CTs' poor bioavailability; (ii) antioxidant action on the gastrointestinal tract; and (iii) interaction with other antioxidant agents. Condensed tannins are alternative dietary antioxidants for ruminants, but further studies should be carried out to elucidate the mechanism underlying the antioxidant activity of each CT source to design effective antioxidant strategies based on the use of CTs in ruminant diets.

Effects of Increasing Doses of Condensed Tannins Extract from Cistus ladanifer L. on In Vitro Ruminal Fermentation and Biohydrogenation.

Cistus ladanifer (rockrose) is a perennial shrub quite abundant in the Mediterranean region, and it is a rich source in secondary compounds such as condensed tannins (CTs). Condensed tannins from C. ladanifer were able to change the ruminal biohydrogenation (BH), increasing the t11-18:1 and c9,t11-18:2 production. However, the adequate conditions of the C. ladanifer CTs used to optimize the production of t11-18:1 and c9,t11-18:2 is not yet known. Thus, we tested the effect of increasing the doses of C. ladanifer CT extract (0, 25, 50, 75 and 100 g/kg dry matter (DM)) on in vitro rumen BH. Five in vitro batch incubations replicates were conducted using an oil supplemented high-concentrate substrate, incubated for 24 h with 6 mL of buffered ruminal fluid. Volatile fatty acids (VFAs) and long chain fatty acids (FA) were analyzed at 0 h and 24 h, and BH of c9-18:1, c9, c12-18:2 and c9, c12, c15-18:3, and BH products yield were computed. Increasing doses of C. ladanifer CTs led to a moderate linear decrease (p < 0.001) of the VFA production (a reduction of 27% with the highest dose compared to control). The disappearance of c9-18:1 and c9,c12-18:2 as well as the production of t11-18:1 and c9, t11:18:2 was not affected by increasing doses of C. ladanifer CTs, and only the disappearance of c9, c12, c15-18:3 suffered a mild linear decrease (a reduction of 24% with the highest dose compared to control). Nevertheless, increasing the C. ladanifer CT dose led to a strong depression of microbial odd and branched fatty acids and of dimethyl acetals production (less than 65% with the highest dose compared to control), which indicates that microbial growth was more inhibited than fermentative and biohydrogenation activities, in a possible adaptative response of microbial population to stress induced to CTs and polyunsaturated fatty acids. The ability of C. ladanifer to modulate the ruminal BH was not verified in the current in vitro experimental conditions, emphasizing the inconsistent BH response to CTs and highlighting the need to continue seeking the optimal conditions for using CTs to improve the fatty acid profile of ruminant fat.

Seasonal Variations of the Nutritive Value and Phytotherapeutic Potential of Cladium mariscus L. (Pohl.) Targeting Ruminant's Production.

In our endeavor to identify salt-tolerant plants with potential veterinary uses in ruminants' production strategies, we focused on Cladium mariscus L. Pohl (sawgrass), due to its high total phenolic and tannin content, anti-radical properties, and ethnomedicinal uses. Aerial parts were collected along the year in Southern Portugal and evaluated for the nutritional profile and in vitro organic matter digestibility (IVOMD), aiming for its use as feed. Acetone extracts were appraised for total contents in phenolics (TPC), flavonoids (TFC), and tannins (CTC), as well as the chemical composition by HPLC-DAD and in vitro antioxidant and anti-inflammatory properties, targeting its exploitation as phytotherapeutic products. Sawgrass biomass has a limited nutritive value, due to its high neutral detergent fiber (NDF; 596-690 g kg-1 dry matter (DM)) and acid detergent fiber (ADF; 330-418 g kg-1 DM) contents, low crude protein (51.8-87.3 g kg-1 DM) and IVOMD (172-317 g kg-1 organic matter (OM)). Despite differences among seasons, the mineral profile was adequate. The extracts were rich in TPC (88-112 mg g-1), CTC (115-169 mg g-1), and TFC (18.5-20.2 mg g-1), and displayed significant antioxidant capacity, particularly in summer and autumn, whilst no seasonal influence was detected for anti-inflammatory properties (30% reduction of nitric oxide production). Eleven phenolics were quantified: chlorogenic, ferulic, and syringic acids were the most abundant, especially in the autumn sample. Overall, despite the low nutritional interest, sawgrass extracts hold the potential as a source of antioxidant and anti-inflammatory phenolic compounds.