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BACKGROUND: Forage inclusion in starters of young dairy calves has become an acceptable strategy in the last decade. To compensate for the lower energy provided by forage, concurrent lipid supplementation can be proposed. However, ruminal microbial activity and forage digestibility may be decreased by lipid supplementation. We hypothesized that the composite effect of forage and lipid supplements may be dependent on forage particle size and the type of lipid supplement. Therefore, we evaluated the effect of long (LP; geometric mean, 4.97 mm) vs. short alfalfa hay particle sizes (SP; geometric mean, 1.26 mm) with either soybean oil (SBO) or palm fatty acids (PLF) as lipid source in a 2 × 2 factorial design with treatments SP-SBO, SP-PLF, LP-SBO, and LP-PLF. Treatments (n = 13 with 6 males and 7 females each) were offered to Holstein calves (3 days old) with equal amounts of lipid (25 g/kg DM) throughout the experimental period. The milk offering scheme (d 1 to 53) was equal for all groups. Data collection continued until 20 d post-weaning. RESULTS: Interaction between forage particle size and lipid supplement was significant for the following readouts: the highest and lowest starter intakes during the pre-weaning period occurred in LP-PLF and LP-SBO, respectively. This was associated with similarly contrasting changes in average daily gain (ADG) during the post-weaning period, body weight at the end of experiment, withers height, digestibility of organic matter and neutral detergent fiber, and blood serum concentrations of glucose, beta-hydroxybutyrate, and insulin during the pre-weaning period. During both pre- and post-weaning periods, the highest and lowest urinary excretion of allantoin and total purine derivatives, representing microbial protein synthesis, were observed in LP-PLF and LP-SBO, respectively, indicating that those diets were most and least favorable for rumen development. Irrespective of forage particle size, supplemental SBO vs. PLF increased serum malondialdehyde as an oxidative stress indicator across periods, increased blood urea nitrogen and feed efficiency in the pre-weaning period, and reduced hip height during the post-weaning period. CONCLUSIONS: It can be concluded that feeding a rumen-inert, mostly saturated fatty acid source with alfalfa hay as long particle size is recommended with view on performance, whereas a combination soybean oil rich in unsaturated fatty acids should not be provided to milk-fed Holstein calves together with long particle forage. Feeding soybean oil and alfalfa hay as long particles is not advisable mainly due to lower starter consumption and impaired development of ruminal function. If dietary supplementation of soybean oil is applied, incorporation of forage as small particles should be preferred to support rumen development.

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Milk proteins are a source of bioactive molecules for calves and humans that may also reflect the physiology and metabolism of dairy cows. Dietary lipid supplements are classically used to modulate the lipid content and composition of bovine milk, with potential impacts on the nutrient's homeostasis and the systemic inflammation of cows that remains to be more explored. This study aimed at identifying discriminant proteins and their associated pathways in twelve Holstein cows (87 ± 7 days in milk), multiparous and non-pregnant, fed for 28 d a diet either, supplemented with 5% DM intake of corn oil and with 50% additional starch from wheat in the concentrate (COS, n = 6) chosen to induce a milk fat depression, or with 3% DM intake of hydrogenated palm oil (HPO, n = 6) known to increase milk fat content. Intake, milk yield and milk composition were measured. On d 27 of the experimental periods, milk and blood samples were collected and label-free quantitative proteomics was performed on proteins extracted from plasma, milk fat globule membrane (MFGM) and skimmed milk (SM). The proteomes from COS and HPO samples were composed of 98, 158 and 70 unique proteins, respectively, in plasma, MFGM and SM. Of these, the combination of a univariate and a multivariate partial least square discriminant analyses reveals that 15 proteins in plasma, 24 in MFGM and 14 in SM signed the differences between COS and HPO diets. The 15 plasma proteins were related to the immune system, acute-phase response, regulation of lipid transport and insulin sensitivity. The 24 MFGM proteins were related to the lipid biosynthetic process and secretion. The 14 SM proteins were linked mainly to immune response, inflammation and lipid transport. This study proposes discriminant milk and plasma proteomes, depending on diet-induced divergence in milk fat secretion, that are related to nutrient homeostasis, inflammation, immunity and lipid metabolism. The present results also suggest a higher state of inflammation with the COS diet.

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This study examines the effects of diets supplemented with various lipids selected to induce divergent milk fat content responses (including a milk fat depression) between dairy cows and goats on plasma lipid composition. The objective was to better understand the mechanisms behind the regulation of milk fat secretion in these two ruminant species. Twelve Holstein cows and 12 Alpine goats were fed a basal diet not supplemented (CTL) or supplemented with corn oil plus wheat starch (COS, 5% DM intake (DMI)), marine algae powder of Schizochytrium sp. (MAP, 1.5% DMI), or hydrogenated palm oil (HPO, 3% DMI), in a replicated 4 × 4 Latin square design, during 28 days. On day 27, blood samples were collected for lipid analysis. Plasma lipid classes were quantified by high-performance thin-layer chromatography, with triacylglycerol (TAG) and free fatty acid (FFA) fractions analysed for FA composition by GLC. Plasma molecular species of TAG and ceramides were determined by HPLC-high-resolution MS and by liquid chromatography-triple quadrupole, respectively. Irrespective of diet, plasma total lipid content was higher in cows than goats (+61%), and TAG concentration was higher in goats than cows (+157%). In cows, conversely to goats, COS increased the trans-10 C18:1 proportion in the free FA (+248%) and the TAG (+195%) fractions. In cows and goats, MAP induced increases in cholesterol esters, cholesterol and phospholipids compared to CTL and changes in the plasma free FA and FA of TAG profiles. In both ruminant species, the concentrations of the lipid fractions were unchanged by HPO compared to CTL. Our results point to species specificities and different diet effects in plasma concentrations and compositions of lipid fractions in cows and goats. These new data highlight how diets, that induce large variations in milk fat secretions, affect the plasma lipid classes available for milk fat synthesis.

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Lipid added as rapeseed or palm oil to the diet of dairy goats over 8 mo of one lactation alters fat secretion and milk fatty acid (FA) and protein composition. In this study, we examined the contribution of mammary gene expression to these changes and included 30 multiparous goats of Norwegian dairy goat breed for a 230-d experimental period, with indoor feeding from 1 to 120 d in milk (DIM), mountain grazing from 120 to 200 DIM, and indoor feeding from 200 to 230 DIM. After an initial period (1-60 DIM) when the control diet was given to all goats, the animals were subdivided into 3 groups of 10 goats. Treatments (60-230 DIM) were basal concentrate (control) alone or supplemented with either 8% (by weight) hydrogenated palm oil enriched with palmitic acid (POFA) or 8% (by weight) rapeseed oil (RSO). Milk was sampled individually from all animals throughout lactation, at 60, 120, 190, and 230 DIM for milk yield and composition. On d 60, 120, 190, and 230, mammary tissue was collected by biopsy to measure mRNA abundance of 19 key genes. None of the 19 genes involved in milk protein, apoptosis, lipid metabolism, transcription factors, and protein of the milk fat globule membrane, as measured by mRNA abundance, were affected by the lipid supplements, although POFA increased milk fat content, and POFA and RSO affected milk FA composition. Over the experimental period (120-230 DIM), the mRNA abundance of 13 of the 19 studied genes was affected by lactation stage. For some genes, expression either gradually increased from 120 to 230 DIM (CSN2, CASP8, CD36, GLUT4) or increased from 120 to 200 and then remained stable (XDH), or decreased (CSN3, G6PD, SREBF1, PPARG1) or increased only at 230 DIM (SCD1, SCD5, ELF3). For a second group of genes (CSN1, LALBA, FABP3, FASN, LPL, MFGE8), expression was stable over the lactation period. Our results suggest that factors other than gene expression, such as substrate availability or posttranscriptional regulation of these genes, could play an important role in the milk fat and FA responses to dietary fat composition in the goat. In conclusion, mammary gene expression in goats was more regulated by stage of lactation than by the dietary treatments applied.

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Recently, the quality of beef has received great attention, and health concerns have been focused on fatty acid composition in relation to dietary requirements. The present work aims to evaluate the effect of omega-3 fatty acids (FA) lipid supplement in beef diet on the nutritional characteristics of Longissimus lumborum (LL) muscle. One hundred and eighty Charolais beef were divided in two groups: the control group (CON) received a basal diet and the second one (TR) an isoenergetic diet containing the omega-3 supplement. Dietary treatment did not affect (p > 0.05) growth performances, carcass characteristics and LL colour indices. Cholesterol content resulted lower (p < 0.001) in LL muscle from TR group than CON. The omega-3 FA and conjugated linoleic acid content were higher (p < 0.001) in LL muscle from TR than CON. As expected, LL muscle from TR group showed an increased value of malondialdehyde than CON during refrigerated storage, anyway, remaining within the threshold value of 1 mg/kg meat. In conclusion, the lipid supplement, rich in omega-3 FA improves the fatty acid profile and decreases cholesterol content of LL muscle. This feeding practice is suggested to enhance the nutritional value of meat from beef reared in intensive condition, improving the consumer's health.

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The objective was to investigate the effects of species (cow vs. goat) and of various dietary lipid supplements, known to modulate milk fat content, on selected metabolites and enzymes in milk and to explore their correlations with performance traits. Twelve Holstein cows and 12 Alpine goats, all multiparous and nonpregnant, and at 86 ± 24.9 and 61 ± 1.8 DIM, respectively, were fed a basal diet (45% forage + 55% concentrate) not supplemented (CTL) or supplemented with corn oil plus wheat starch [COS, 5% of diet dry matter (DM)], marine algae powder (MAP, 1.5% of diet DM), or hydrogenated palm oil (HPO, 3% of diet DM) in a replicated 4 × 4 Latin square design with 28-d experimental periods. Intake, milk production and composition, milk fatty acid profile, and plasma metabolite concentrations were previously reported. Concentrations of 9 milk metabolites [ß-hydroxybutyrate (BHB), glucose, glucose-6-phosphate, isocitrate, choline, glutamate, urea, cholesterol, and free amino groups] and 2 milk enzyme activities (alkaline phosphatase and lactate dehydrogenase) were measured on d 24 of each experimental period. Dairy performance data showed marked species and diet effects on milk fat content. Irrespective of diet, cow milk was richer in alkaline phosphatase and glucose compared with goat milk (16 and 3 times more, respectively), whereas goat milk had greater urea and glucose-6-phosphate concentrations compared with cow milk (1.9 and 5.3 times more, respectively). In cows, COS decreased milk BHB and choline (-25 and -43%, respectively) compared with CTL, whereas no effects were observed in goats. The COS and MAP diets increased milk isocitrate compared with CTL in cows, but COS decreased isocitrate concentrations in goat milk. Milk choline was correlated with milk fat content in cows (Spearman r, rS = +0.73) and goats (rs = +0.58), and lactate dehydrogenase activity was correlated with milk somatic cell count (rs = +0.66) in cows but not in goats. We provide evidence of different milk metabolite responses according to species and diets. Metabolites and enzymes secreted in milk may be indicators of specificities of lipid metabolism among ruminant species and may contribute to a better understanding of mechanisms regulating milk fat secretion. Changes in the concentrations of some metabolites considered minor components of milk may be valuable diagnostic tools of mammary gland and animal metabolism as well as of milk processing characteristics.

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A direct comparison of cow and goat performance and milk fatty acid (FA) responses to diets that either induce milk fat depression or increase milk fat content in cows suggests species-specific regulation of lipid metabolism, including mammary lipogenesis. This experiment was conducted to highlight potential mechanisms responsible for the differences in mammary lipogenesis due to diet and ruminant species. Twelve Holstein cows and 12 Alpine goats were fed a basal diet containing no additional lipid (CTL) or a similar diet supplemented with corn oil [5% dry matter intake (DMI)] and wheat starch (COS), marine algae powder (MAP; 1.5% DMI), or hydrogenated palm oil (HPO; 3% DMI), according to a 4 × 4 Latin square design with 28-d experimental periods. Milk yield, milk composition, FA profile, and secretions were measured. On d 27 of each experimental period, the mRNA abundance of 21 genes involved in lipid metabolism or enzyme activities or both were measured in mammary tissue sampled by biopsy. The results showed significant differences in the milk fat response of cows and goats to the dietary treatments. In cows, fat content was lowered by COS (-45%) and MAP (-22%) and increased by HPO (+13%) compared with CTL, and in goats only MAP had an effect compared with CTL, with a decrease of 15%. In both species, COS and MAP lowered the yields (mmol/d per kilogram of body weight) of C16 FA in goats but not in cows, and the >C16 FA yield decreased with MAP in both species. Supplementation of HPO increased the yield of milk C16 FA (mmol/d per kilogram of body weight) in cows. These variations in milk fat content and FA secretion were not associated with modifications in the mammary expression of 21 genes involved in major lipid pathways, except for 3 transcription factors: PPARA, INSIG1, and SP1. This absence of large changes might be due to post-transcriptional regulation of these genes and related to the time of sampling of the mammary tissue relative to the previous meal and milking or to differences in the availability of substrate for the corresponding proteins. However, the abundance of 14 mRNA among the 21 encoding for genes studied in the mammary gland was significantly different among species, with 5 more abundant in cows (FADS3, ACSL1, PPARA, LXRA, and PPARG1) and 10 more abundant in goats (FASN, CD36, FABP3, LPL, GPAM, LPIN1, CSN2, MFGE8, and INSIG1). These species specificities of mammary lipid metabolism require further investigation.

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Abstract Background: Dietary linoleic (LA) and alpha-linolenic (LN) acids are extensively isomerized and hydrogenated by rumen microbes, and this activity can further contribute to the fatty acid profile of ruminant- derived food products. Objective: To evaluate the effects of LA:LN ratio in lipid supplements on the rumen biohydrogenation kinetics of LA and LN, as well as on the trans-vaccenic acid (VA) production, using an in vitro system. Methods: Rumen fluid was collected from a fistulated steer, diluted with incubation buffer, and then incubated with 500 mg of kikuyu grass (Cenchrus clandestinus) supplemented with 16.3 mg of different LA:LN mixtures (100:0, 75:25, 50:50, 25:75 or 0:100). Incubations were performed in triplicate for a period of 0, 2, 4, 6, 8 or 16 hours. Differences between treatments were evaluated in a completely randomized design. Alternatively, computational chemistry was used to determine the changes in the Gibbs free energy (ΔGrxn) at 39 °C for the principal steps of LA and LN ruminal biohydrogenation. Results: Partial replacement of LA by LN decreased the VA concentration and its accumulation rate; it also increased the stearic acid concentration and the rates of transfer from LA to conjugated linoleic acid (CLA), and from CLA to VA. The conversion from CLA to VA (ΔGrxn = -2.65 kJ/mol) was more spontaneous than that from trans-11, cis-15 octadecadienoic acid (TA) to VA (ΔGrxn = -0.29 kJ/mol). Conclusion: The LA:LN ratio in lipids can modulate LA and LN biohydrogenation (BH) kinetics, as well as the VA production in the rumen.

Resumen Antecedentes: los ácidos linoleico (LA) y alfa-linolénico (LN) de la dieta son extensivamente isomerizados y biohidrogenados por los microorganismos ruminales, lo cual puede contribuir al perfil de ácidos grasos de los productos derivados de rumiantes. Objetivo: evaluar el efecto de la relación LA:LN en suplementos lipídicos sobre la cinética de biohidrogenación ruminal del LA y LN, como también sobre la producción del ácido trans-vaccénico (VA), usando un sistema in vitro. Métodos: se colectó fluido ruminal de un toro fistulado, el cual fue diluido con buffer de incubación y posteriormente incubado con 500 mg de pasto kikuyo (Cenchrus clandestinus) suplementado con 16,3 mg de diferentes mezclas de LA:LN (100:0, 75:25, 50:50, 25:75, o 0:100). Las incubaciones fueron desarrolladas en triplicado durante 0, 2, 4, 6, 8 o 16 horas. Diferencias entre tratamientos fueron evaluadas mediante un modelo completamente al azar. Alternativamente, se determinaron los cambios en energía libre de Gibbs ( Δ Grxn) a 39 °C para los pasos principales de la biohidrogenación del LA y LN, usando química computacional. Resultados: la sustitución parcial de LA por LN disminuyó la concentración de VA y su tasa de acumulación, como también incrementó la concentración de ácido esteárico y las tasas de transferencia de LA para ácido linoleico conjugado (CLA) y de CLA para VA. La conversión de CLA para VA ( Δ Grxn = -2,65 kJ/mol) fue más espontánea que la conversión del ácido trans-11, cis-15 octadecadienóico (TA) para VA ( Δ Grxn = -0,29 kJ/mol). Conclusiones: la relación LA:LN en lípidos puede modular la cinética de biohidrogenación (BH) del LA y LN y la producción de VA en el rumen.

Resumo Antecedêntes: o ácido linoleico (LA) e alfa-linolênico (LN) da dieta, são extensivamente isomerizados e biohidrogenados pelos microorganismos do rúmen, o que pode contribuir ao perfil de ácidos graxos dos produtos derivados de ruminantes. Objetivo: avaliar o efeito da relação LA:LN em suplementos lipídicos sobre a cinética de biohidrogenação ruminal do LA e LN como também sobre a produção do ácido trans- vaccênico (VA), utilizando um sistema in vitro. Métodos: coletou-se fluido ruminal de um novilho fistulado, o qual foi diluído com tampão de incubação e, em seguida, incubado com 500 mg de pasto kikuyu (Cenchrus clandestinus) suplementado com 16,3 mg de diferentes misturas LA:LN (100:0, 75:25 , 50:50, 25:75 ou 0:100). As incubações foram desenvolvidas em triplicata, durante 0, 2, 4, 6, 8 ou 16 horas. Diferenças entre tratamentos foram avaliadas utilizando-se um delineamento inteiramente casualizado. Alternativamente, foram determinadas as mudanças em energia livre de Gibbs ( Δ Grxn) a 39 °C para as principais etapas da biohidrogenação do LA e LN, utilizando-se química computacional. Resultados: a substituição parcial de LA por LN diminuiu a concentração de VA e sua taxa de acumulação, como também aumentou a concentração de ácido esteárico e as taxas de transferência do LA para o ácido linoleico conjugado (CLA) e do CLA para VA. A conversão do CLA para VA ( Δ Grxn = -2,65 kJ/ mol) foi mais espontânea que a conversão do ácido trans-11, cis-15 octadecadienóico (TA) para VA ( Δ Grxn = -0,29 kJ/mol). Conclusões: a relação LA:LN em lipídeos pode modular a cinética de biohidrogenação (BH) do LA e LN e a produção de VA no rúmen.

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A direct comparative study of dairy cows and goats was performed to characterize the animal performance and milk fatty acid (FA) responses to 2 types of diets that induce milk fat depression in cows as well as a diet that increases milk fat content in cows but for which the effects in goats are either absent or unknown. Twelve Holstein cows and 12 Alpine goats, all multiparous, nonpregnant, and at 86 ± 24.9 and 61 ± 1.8 DIM, respectively, were allocated to 1 of 4 groups and fed diets containing no additional lipid (CTL) or diets supplemented with corn oil [5% dry matter intake (DMI)] and wheat starch (COS), marine algae powder (MAP; 1.5% DMI), or hydrogenated palm oil (HPO; 3% DMI), according to a 4 × 4 Latin square design with 28-d experimental periods. Dietary treatments had no significant effects on milk yield and DMI in both species, except for COS in cows, which decreased DMI by 17%. In cows, milk fat content was lowered by COS (-45%) and MAP (-22%) and increased by HPO (13%) compared with CTL, and in goats only MAP had an effect compared with CTL by decreasing milk fat content by 15%. In both species, COS and MAP lowered the yields (mmol/d per kg of BW) of C16 FA in goats, but not in cows, and the >C16 FA yield decreased with MAP in both species. HPO supplementation increased the milk yield of C16 FA in cows. Compared with CTL, COS induced an increase of trans-10,cis-12 conjugated linoleic acid by 18 fold in cows and 7 fold in goats and of trans-10 18:1 by 13 fold in cows and 3 fold in goats. Moreover, other conjugated linoleic acid isomers, such as trans-10,trans-12 and trans-7,cis-9, were increased to a greater extent in cows (8 and 4 fold, respectively) compared with goats (4 and 2 fold, respectively) on the COS treatment. In both species, the responses to MAP were characterized by a decrease in the milk concentration of 18:0 (3 fold, on average) and cis-9 18:1 (2 fold, on average) combined with a 3-fold increase in the total trans 18:1, with an increase in trans-10 18:1 only observed in cows. Compared with CTL, the response to HPO was distinguished by an increase in 16:0 (10%) in cows. This comparative study clearly demonstrated that each ruminant species responds differently to COS and HPO treatments, whereas MAP caused similar effects, and that goats are less sensitive than cows to diets that induce a shift from the trans-11 toward the trans-10 ruminal pathways.

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PURPOSE: To compare the prophylactic efficacy of single application of lipid and non-lipid containing tear supplements, prior to exposure of symptomatic dry eye subjects to a simulated adverse environment. METHODS: Thirty subjects with mild-to-moderate dry eye symptoms participated in the prospective, randomised, double-masked, paired-eye trial. A lipomimetic drop (Systane® Balance) was applied to one eye (randomised), and a non-lipid containing drop (Systane® Ultra) applied simultaneously to the contralateral eye. Subjects were subsequently exposed to a validated simulated adverse environment model created by a standing fan directed towards the eye, at a distance of 1m, for 2.5min. Low contrast glare acuity, lipid layer grade (LLG), non-invasive tear break-up time (NIBUT), temperature variation factor (TVF), and tear meniscus height (TMH) were evaluated at baseline, following eye drop instillation and following simulated adverse environment exposure. RESULTS: Both therapies resulted in increased NIBUT (both p<0.001), and prevented its decline below baseline with simulated adverse environment exposure (both p>0.05). However, only the lipomimetic drop increased LLG (p<0.001) and precluded its fall below baseline post-adverse environment exposure (p=0.15). Furthermore, post-instillation and post-exposure LLGs and NIBUT were significantly higher in the lipomimetic group (all p<0.05). No significant changes were observed in glare acuity, TVF and TMH (all p>0.05). More subjects (67%) reported greater ocular comfort in the eye receiving the lipomimetic. CONCLUSIONS: Single application of both lipid and non-lipid containing eye drops conferred protective effects against exposure to adverse environmental conditions in subjects with mild-to-moderate dry eye, although the lipomimetic demonstrated superior prophylactic efficacy.

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We previously reported that 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (PLAG) accelerates hematopoiesis and has an improving effect on animal disease models such as sepsis and asthma. The effects of PLAG supplementation on immune modulation were assessed in healthy men and women. The objective was to evaluate the effects of PLAG supplementation on immune regulatory functions such as activities of immune cells and cytokine production. A randomized double blind placebo-controlled trial was conducted. Seventy-five participants were assigned to one of two groups; all participants had an appropriate number of white blood cells on the testing day. The PLAG group (n=27) received oral PLAG supplements and the control group (n=22) received oral soybean oil supplements. IL-4 and IL-6 production by peripheral blood mononuclear cells (PBMC) were lower (p<0.001 and p<0.001, respectively) with PLAG than with soybean oil. However, the production of IL-2 and IFN-γ by PBMC was unaltered with PLAG supplementation. The B cell proliferation decreased significantly in the PLAG group compared to the soybean oil control (p<0.05). The intake of PLAG in healthy adults for 4 weeks was deemed safe. These data suggest that PLAG has an immunomodulatory function that inhibits the excessive immune activity of immunological disorders such as atopic and autoimmune diseases. PLAG could improve the condition of these diseases safely as a health food supplement.

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We previously reported that 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (PLAG) accelerates hematopoiesis and has an improving effect on animal disease models such as sepsis and asthma. The effects of PLAG supplementation on immune modulation were assessed in healthy men and women. The objective was to evaluate the effects of PLAG supplementation on immune regulatory functions such as activities of immune cells and cytokine production. A randomized double blind placebo-controlled trial was conducted. Seventy-five participants were assigned to one of two groups; all participants had an appropriate number of white blood cells on the testing day. The PLAG group (n=27) received oral PLAG supplements and the control group (n=22) received oral soybean oil supplements. IL-4 and IL-6 production by peripheral blood mononuclear cells (PBMC) were lower (p<0.001 and p<0.001, respectively) with PLAG than with soybean oil. However, the production of IL-2 and IFN-gamma by PBMC was unaltered with PLAG supplementation. The B cell proliferation decreased significantly in the PLAG group compared to the soybean oil control (p<0.05). The intake of PLAG in healthy adults for 4 weeks was deemed safe. These data suggest that PLAG has an immunomodulatory function that inhibits the excessive immune activity of immunological disorders such as atopic and autoimmune diseases. PLAG could improve the condition of these diseases safely as a health food supplement.

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