Essential oil supplementation in milk replacers: short- and long-term impacts on feed efficiency, the faecal microbiota and the plasma metabolome in dairy calves.

Early supplementation with oregano essential oil (EO) in milk replacer (MR) may improve growth, immune responses, the microbiota and the metabolome in dairy calves during pre-weaning and in adulthood. Sixteen female dairy calves (3 days of age) were divided in two groups (n = 8/group): the control group (no EO) and the EO group (0.23 ml of EO in MR during 45 days). After weaning, calves were kept in a feedlot and fed ad libitum. The animals were weighed, and blood and faecal samples were collected on days 3 (T0), 45 (T1) and 370 (T2) to measure the biochemical profile and characterise peripheral blood mononuclear cells (PBMCs; CD4+, CD8+, CD14+, CD21+ and WC1+), the metabolome and microbiota composition. The EO group only had greater average daily weight gain during the suckling (EO supplementation) period (P = 0.030). The EO group showed higher average CD14+ population (monocytes) values, a lower abundance of Ruminococcaceae UCG-014, Faecalibacterium, Blautia and Alloprevotella and increased abundances of Allistipes and Akkermansia. The modification of some metabolites in plasma, such as butyric acid, 3-indole-propionic acid and succinic acid, particularly at T1, are consistent with intestinal microbiota changes. The data suggest that early EO supplementation increases feed efficiency only during the suckling period with notable changes in the microbiota and plasma metabolome; however, not all of these changes can be considered desirable from a gut health point of view. Additional research studies is required to demonstrate that EOs are a viable natural alternative to antibiotics for improving calf growth performance and health.

Dietary Administration of L-Carnitine During the Fattening Period of Early Feed Restricted Lambs Modifies Ruminal Fermentation but Does Not Improve Feed Efficiency.

Early feed restriction of lambs may program animals to achieve reduced feed efficiency traits as a consequence of permanent mitochondrial dysfunction. The hypothesis at the background of the present study is that dietary administration of L-Carnitine (a compound that promotes the activation and transportation of fatty acids into the mitochondria) during the fattening period of early feed restricted lambs can: (a) improve the biochemical profile of early feed restricted lambs, (b) improve feed efficiency, (c) modulate the ruminal and intestinal microbiota, and (d) induce changes in the gastrointestinal mucosa, including the immune status. Twenty-two newborn male Merino lambs were raised under natural conditions but separated from the dams for 9 h daily to allow feed restriction during the suckling period. At weaning, lambs were assigned to a control group being fed ad libitum a complete pelleted diet during the fattening phase (CTRL, n = 11), whereas the second group (CARN, n = 11) received the same diet supplemented with 3 g of L-Carnitine/kg diet. The results revealed that even though L-Carnitine was absorbed, feed efficiency was not modified by dietary L-Carnitine during the fattening period (residual feed intake, p > 0.05), whereas ruminal fermentation was improved [total short-chain fatty acids (SCFAs), 113 vs. 154 mmol/l; p = 0.036]. Moreover, a trend toward increased concentration of butyrate in the ileal content (0.568 vs. 1.194 mmol/100 ml SCFA; p = 0.074) was observed. Other effects, such as reduced heart weight, lower levels of markers related to muscle metabolism or damage, improved renal function, and increased ureagenesis, were detected in the CARN group. Limited changes in the microbiota were also detected. These findings suggest that L-Carnitine may improve ruminal fermentation parameters and maintain both the balance of gut microbiota and the health of the animals. However, the improved ruminal fermentation and the consequent greater accumulation of intramuscular fat might have hidden the effects caused by the ability of dietary L-Carnitine to increase fatty acid oxidation at the mitochondrial level. This would explain the lack of effects of L-Carnitine supplementation on feed efficiency and points toward the need of testing lower doses, probably in the context of animals being fed in excess non-protein nitrogen.

Dietary supplemental plant oils reduce methanogenesis from anaerobic microbial fermentation in the rumen.

Ruminants contribute to the emissions of greenhouse gases, in particular methane, due to the microbial anaerobic fermentation of feed in the rumen. The rumen simulation technique was used to investigate the effects of the addition of different supplemental plant oils to a high concentrate diet on ruminal fermentation and microbial community composition. The control (CTR) diet was a high-concentrate total mixed ration with no supplemental oil. The other experimental diets were supplemented with olive (OLV), sunflower (SFL) or linseed (LNS) oils at 6%. Rumen digesta was used to inoculate the fermenters, and four fermentation units were used per treatment. Fermentation end-products, extent of feed degradation and composition of the microbial community (qPCR) in digesta were determined. Compared with the CTR diet, the addition of plant oils had no significant (P > 0.05) effect on ruminal pH, substrate degradation, total volatile fatty acids or microbial protein synthesis. Gas production from the fermentation of starch or cellulose were decreased by oil supplementation. Methane production was reduced by 21-28% (P < 0.001), propionate production was increased (P < 0.01), and butyrate and ammonia outputs and the acetate to propionate ratio were decreased (P < 0.001) with oil-supplemented diets. Addition of 6% OLV and LNS reduced (P < 0.05) copy numbers of total bacteria relative to the control. In conclusion, the supplementation of ruminant diets with plant oils, in particular from sunflower or linseed, causes some favorable effects on the fermentation processes. The addition of vegetable oils to ruminant mixed rations will reduce methane production increasing the formation of propionic acid without affecting the digestion of feed in the rumen. Adding vegetable fats to ruminant diets seems to be a suitable approach to decrease methane emissions, a relevant cleaner effect that may contribute to alleviate the environmental impact of ruminant production.

Effects of supplemental plant oils on rumen bacterial community profile and digesta fatty acid composition in a continuous culture system (RUSITEC).

Lipid supplementation of ruminant diets may trigger changes in the ruminal microbiota and in anaerobic digestion. Changes in the bacterial community composition and in the fatty acid hydrogenation caused by the addition of different supplemental plant oils to a high concentrate diet were investigated in vitro using RUSITEC (rumen simulation technique) fermenters. The control (CTR) diet was a high-concentrate total mixed ration for dairy sheep, with no supplementary oil. The other experimental diets were supplemented with olive (OLV), sunflower (SFL) or linseed (LNS) oils at 6% (dry matter basis). Four RUSITEC fermenters were used for each experimental diet, all inoculated with rumen digesta of sheep. Extent of dry matter and fat degradation, composition of the bacterial community and long-chain fatty acids in digesta were determined. The addition of plant oils increased (P < 0.001) apparent degradation of fat in the fermenters, whereas fermentation kinetics (gas production and average fermentation rate) were lower (P < 0.05) with the LNS than with the CTR diet. Hydrogenation of C18 unsaturated fatty acids (P < 0.05), in particular that of oleic acid (P < 0.001), and stearic acid proportion (P < 0.001) were reduced, and oleic acid proportion was increased (P < 0.001) with all oil supplements. Addition of OLV decreased linoleic and LNS increased α-linolenic (P < 0.001), whereas conjugated linoleic was increased with SFL oil (P = 0.025) and vaccenic increased with both SFL and LNS oils (P = 0.008). Addition of 6% OLV and LNS reduced (P < 0.05) microbial community diversity and quantity of total bacteria relative to the control. Some specific microbial groups were affected (P < 0.001) by oil addition, with less relative abundance of Clostridiales and Actinobacteria and increased Bacteroidales, Aeromonadales and Lactobacillales species. In conclusion, the supplementation of high-concentrate ruminant diets with plant oils, in particular from sunflower or linseed, causes shifts in the rumen microbiota and fatty acid hydrogenation in the rumen increasing the formation of vaccenic and conjugated linoleic acids.

Effects of dietary astaxanthin supplementation on the oxidative stability of meat from suckling lambs fed a commercial milk-replacer containing butylated hydroxytoluene.

Meat colour and lipid oxidative stability can be improved by adding antioxidants to animal diet. This study investigated the effects of the addition of astaxanthin to a butylated hydroxytoluene (BHT)-containing commercial milk-replacer, at a rate of 25 mg of astaxanthin/kg of milk-replacer powder, on suckling lamb meat quality. Twenty newborn (2 day old) lambs allocated to individual pens were artificially reared for 22 days. Ten lambs (Control) were fed a commercial milk-replacer and the other ten (Astaxanthin) received the same milk-replacer but included astaxanthin. After the feeding trial, meat and fat colour, astaxanthin and BHT levels in meat, oxidative stability in refrigerated and frozen raw meat and refrigerated cooked meat, and meat volatiles in cooked meat were determined. Astaxanthin in artificially reared suckling lambs at the levels used reduced the accumulation of BHT in the meat, slightly affected meat colour, by reducing meat lightness and increasing meat and fat redness, and increased the lipid stability of frozen meat.

Effect of hop (Humulus lupulus L.) inclusion in the diet for fattening lambs on animal performance, ruminal characteristics and meat quality.

Thirty male merino lambs were fed with a pelleted total mixed ration (TMR) alone or supplemented with hop (Humulus lupulus L.) cones at two different doses (1.5 and 3.0 g hop cones/kg pelleted TMR, respectively), to study the effects of this dietary source of antioxidants on animal performance, ruminal parameters and meat quality attributes. The results showed that dietary supplementation with hop cones decreased lambs' growth rate (P < 0.05) due to a shift in ruminal fermentation, towards a more acetic and less propionic acid production (P < 0.05). These changes in animal growth rate might have promoted microstructural modifications in the quantity and size of muscle fibres, thereby inducing the differences observed in meat chemical composition, colour and texture (P < 0.05), regardless of the lack of differences in meat antioxidant status (P > 0.10).

Effect of Sunflower and Marine Oils on Ruminal Microbiota, In vitro Fermentation and Digesta Fatty Acid Profile.

This study using the rumen simulation technique (RUSITEC) investigated the changes in the ruminal microbiota and anaerobic fermentation in response to the addition of different lipid supplements to a ruminant diet. A basal diet with no oil added was the control, and the treatment diets were supplemented with sunflower oil (2%) only, or sunflower oil (2%) in combination with fish oil (1%) or algae oil (1%). Four fermentation units were used per treatment. RUSITEC fermenters were inoculated with rumen digesta. Substrate degradation, fermentation end-products (volatile fatty acids, lactate, gas, methane, and ammonia), and microbial protein synthesis were determined. Fatty acid profiles and microbial community composition were evaluated in digesta samples. Numbers of representative bacterial species and microbial groups were determined using qPCR. Microbial composition and diversity were based on T-RFLP spectra. The addition of oils had no effect on substrate degradation or microbial protein synthesis. Differences among diets in neutral detergent fiber degradation were not significant (P = 0.132), but the contrast comparing oil-supplemented diets with the control was significant (P = 0.039). Methane production was reduced (P < 0.05) with all oil supplements. Propionate production was increased when diets containing oil were fermented. Compared with the control, the addition of algae oil decreased the percentage C18:3 c9c12c15 in rumen digesta, and that of C18:2 c9t11 was increased when the control diet was supplemented with any oil. Marine oils decreased the hydrogenation of C18 unsaturated fatty acids. Microbial diversity was not affected by oil supplementation. Cluster analysis showed that diets with additional fish or algae oils formed a group separated from the sunflower oil diet. Supplementation with marine oils decreased the numbers of Butyrivibrio producers of stearic acid, and affected the numbers of protozoa, methanogens, Selenomonas ruminantium and Streptococcus bovis, but not total bacteria. In conclusion, there is a potential to manipulate the rumen fermentation and microbiota with the addition of sunflower, fish or algae oils to ruminant diets at appropriate concentrations. Specifically, supplementation of ruminant mixed rations with marine oils will reduce methane production, the acetate to propionate ratio and the fatty acid hydrogenation in the rumen.

Effect of dietary supplementation with carnosic acid or vitamin E on animal performance, haematological and immunological characteristics of artificially reared suckling lambs before and after road transport.

To elucidate the influence of dietary carnosic acid (CA) and vitamin E on animal performance, immune response indicators and haematological parameters before and after transport stress, 24 lambs were individually fed ad libitum with milk replacer (MR) using an auto-feeder. Once daily the lambs received MR alone (Group CON, n = 8), MR + 0.096 g CA/kg live weight (LW) (Group CARN, n = 8) or MR + 0.024 g of α-tocopheryl acetate per kg LW (Group VitE, n = 8). After reaching the target slaughter weight (12 ± 0.5 kg), blood samples were collected to measure haematological and immunological parameters. Then, lambs were subjected to 4-h road transport and blood samples were collected again for haematological assessment. The animals were subsequently slaughtered. Before road transport, dietary CA supplementation promoted a descent of circulating white blood cells (WBC), red blood cells (RBC), haematocrit and haemoglobin concentration when compared with Groups CON and VitE (p < 0.05), but it did not affect production of cytokines by blood mononuclear cells. Road transport did not affect either RBC or haematocrit significantly. Nevertheless, transport affected leucocyte profile similarly in all the treatments, increasing granulocytes and monocytes proportions and decreasing lymphocytes. In contrast, after transport, WBC was increased in Group CARN, reaching similar values than Groups CON and VitE. However, under conditions of the present study, those modifications did not influence animal performance or immunity parameters of artificially reared suckling lambs.

Effect of dietary carnosic acid on the fatty acid profile and flavour stability of meat from fattening lambs.

Thirty-two lambs were fed with barley straw supplemented by a concentrate alone, or a concentrate enriched with either vitamin E (VITE006: 0.6 g kg(-1) feed concentrate) or carnosic acid (CARN006: 0.6 g kg(-1) feed concentrate; or CARN012: 1.2 g kg(-1) feed concentrate). In order to elucidate the influence of the dietary supplementation of carnosic compared with a reference diet antioxidant (vitamin E), the animals were slaughtered and the longissimus thoracis were lyophilised to determine the FAs profile and the phenolic compounds. In addition, longissimus lumborum slices were stored in a modified atmosphere package for 3 days and then grilled to determine volatile compounds. Dietary carnosic acid did not modify the FAs profile, but had a clear effect on the production of volatile compounds, in a dose-dependent manner. These results have implications for the food industry, since dietary carnosic acid seems to extend the shelf life of lamb meat.

Meat texture and antioxidant status are improved when carnosic acid is included in the diet of fattening lambs.

Thirty-two Merino lambs fed barley straw and a concentrate alone (CONTROL group) or enriched with carnosic acid [0.6 g kg(-1) dry matter (DM), CARN006 group; 1.2 g kg(-1) DM, CARN012 group] or vitamin E (0.6 g kg(-1) DM, VITE006 group) were used to assess the effect of these antioxidant compounds on meat quality. After being fed the experimental diets for at least 5 weeks, the animals were slaughtered with the 25 kg intended body weight and the different muscles (longissimus lumborum; LL, gluteus medius; GM) were sliced and kept refrigerated under modified atmosphere packaging during 0, 7 and 14 days. The results indicate that carnosic acid seemed to be useful to delay lipid peroxidation in a medium colour-stable muscle such as GM, but this effect was lower than that observed when vitamin E was supplemented to fattening lambs. On the contrary, meat texture and protection against cholesterol oxidation were equally improved with both compounds.