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.

Secretory IgA as Biomarker for Gastrointestinal Nematodes Natural Infection in Different Breed Sheep.

Specific IgA antibody has been shown to play an important role in resistance to gastrointestinal nematode (GIN) infections in sheep, particularly in Teladorsagia circumcincta parasitosis. In some breeds, negative associations have been shown between IgA levels and worm burden in experimentally infected sheep. In the present study, we have studied the relationship between IgA levels in naturally infected sheep (582 ewes in total; 193 younger than one year old and 389 older than one year old) and fecal egg count (FEC) in the Assaf, Castellana, and Churra breeds. ELISA assays were performed to measure IgA levels against the somatic antigen of T. circumcincta third larval stage (L3) and a 203-amino-acid fragment of the protein disulfide isomerase from the same GIN species. A multilevel random intercept model was developed to predict the infection risk according to age or breed. Spearman's correlation rank was used for statistical analysis. The prediction model showed that breed was not an influential factor in this study, although the Assaf breed could be considered slightly more susceptible than the others. In addition, age affected the infection risk, with the young ewes more susceptible to infection than the adult groups, except for the Castellana breed, whose risk of infection was similar at all ages. The most significant positive association was found between FEC and IgA measured in the nasal secretions of young ewes using both antigens (Rho = 0.5; p = 0.00); the correlation of FEC with IgA in serum was moderately significant (Rho = 0.306; p = 0.00). Comparing both antigens, the protein disulfide isomerase antigen was less reactive than the somatic antigen from L3. In conclusion, under natural conditions, specific IgA against GIN was positively associated with FEC in sheep, with nasal secretions from young animals being the sample where this association is stronger, which, therefore, could be used as a marker of infection in further studies.

Effects of Thymbra capitata essential oil on in vitro fermentation end-products and ruminal bacterial communities.

An in vitro trial was carried out to investigate the effects of natural Thymbra capitata essential oil (NEO) and its main compounds [including carvacrol, p-cymene, γ-terpinene given alone or in a synthetic combination (SEO)] on ruminal fermentation and the bacterial community using batch cultures inoculated with ruminal digesta and incubating two different basal diets [high-forage (F) and high-concentrate (C) diet]. After 24 h of incubation, primary fermentation end-products [gas, methane, volatile fatty acids (VFAs) and ammonia] and rumen microbial diversity were determined. NEO reduced the total VFA concentration (P < 0.05) only in the C diet. In contrast, SEO and carvacrol decreased the total VFA concentration (P < 0.05) only in the F diet. Methane production was not affected (P > 0.05) by any of the experimental treatments or diets evaluated. Microbial diversity analysis showed only a moderate effect of carvacrol and SEO on 13 genera, including, mainly, Atopobium and Blautia (involved in subacute ruminal acidosis) or Candidatus Saccharimonas (related to laminitis). In conclusion, T. capitata EO has a limited potential to attain nutritional or environmental benefits, but further research should be carried out to clarify its effects on animal health and microbial food safety.

The Effects of Aflatoxin B1 Intake in Assaf Dairy Ewes on Aflatoxin M1 Excretion, Milk Yield, Haematology and Biochemical Profile.

The aim of this study was to investigate the in vivo transfer of aflatoxin B1 (AFB1) to Assaf ewes' milk (aflatoxin M1, AFM1) and its effect on animal performance and health. Thirty Assaf ewes were allocated to three groups (C, L, H), and received a different individual daily dose of AFB1 (0, 40 and 80 µg) for 13 days. Milk (days 1, 2, 3, 4, 7, 14, 16 and 18) and blood (days 1, 7, 14 and 18) samples were collected. Milk yield, composition (except protein) and somatic cell counts (SCC) were not affected by AFB1 intake (p > 0.05). Haemoglobin concentration increased (p < 0.05) and haematocrit and alanine aminotransferase levels tended to increase (p < 0.10) in group H on day 14. AFM1 excretion was highly variable and detected in L and H animals from days 1 to 16 (3 days increase, 10 days steady-state, 3 days clearance). Carry-over rate (0.23%) was significantly higher in L (0.22-0.34%) than in H (0.16-0.19%) animals (p < 0.05). AFB1 daily doses of 40 to 80 µg do not impair milk yield; however, it may start affecting animals' health. Milk AFM1 depends mainly on the AFB1 intake whereas carryover rate is positively influenced by the level of milk production.

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.

The Role of Feed Restriction on DNA Methylation, Feed Efficiency, Metabolome, Biochemical Profile, and Progesterone Patterns in the Female Filial Generation (F1) Obtained From Early Feed Restricted Ewes (F0).

Deficient management of replacement animals in the farm during early developmental windows may promote adverse programming effects on reproductive traits and subsequent transmission to the next generation. In this sense, DNA methylation profiles allow researchers to decode epigenetic regulation mechanisms in mammals and identify novel candidate genes correlated with phenotype differences in both dams and offspring. Therefore, improving knowledge in the field of epigenetics and intergenerational effects caused by prenatal and postnatal early nutritional events (e.g., feed restriction) is crucial for refining strategies dedicated to animal breeding. In this study, we determined differences in the global blood methylation patterns, biochemical profile, and metabolome of ewe lambs (F1) born from either early feed restricted dams (F0-RES) or fed ad libitum (F0-ADL). Our data show that functional categories such as those related to cellular processes, phosphorylation, nervous system, immunity response, or reproductive function were enriched significantly in the F1-RES lambs due to differences in the methylation of genes in these categories. These F1-RES lambs did not show differences in feed efficiency during the replacement period but presented higher levels of insulin and triglycerides and reduced concentration of progesterone, whereas the metabolome profile demonstrated variations in the bile acid composition when compared with the F1-ADL lambs. Taken together, all these results suggest that intergenerational effects caused by early feed restriction of dams (F0) may persist in the F1 female lambs with negative consequences on genes involved in cellular processes and reproductive traits.

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).

Programming Merino lambs by early feed restriction reduces growth rates and increases fat accretion during the fattening period with no effect on meat quality traits.

The objective of the study was to determine the effect of early feed restriction on carcass and meat quality traits in Merino lambs. Twenty-four male Merino lambs (4.81±0.256kg) were assigned to two groups (12 animals in each group). The first group (ad libitum, ADL) was kept permanently with the dams. The second group (restricted, RES) was milk-restricted. After weaning (15kg), daily feed intake of all lambs was identical, but a lower gain: feed ratio was recorded in the RES group (0.320 vs. 0.261; P<0.001). After animals were slaughtered (27kg), lower ultimate pH and reduced yellow colouration of subcutaneous fat was recorded in meat from the RES lambs when compared to ADL group, whereas a trend towards a significant increment of intramuscular fat content and no effect on meat quality traits was observed in the RES lambs. Early feed restriction may be useful to increase intramuscular fat accretion in Merino lambs during the fattening period.

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.