Supplementation with a blend based on micro-encapsulated carvacrol, thymol, and cinnamaldehyde in lambs feed inhibits immune cells and improves growth performance
Introduction
In Brazil, the first sheep that produced milk were introduced around 1992; these were derived from the Lacaune breed from France, and they demonstrated excellent adaptation to Brazilian feed and climate (Brito et al., 2006). A major challenge with raising ruminants is their high mortality in the post-weaning period, especially among dairy animals (Budzynska and Weary 2008). This is because the lambs are removed from their mothers (i.e., weaned) very early and the digestive system is not completely adapted to receive solid food only. Until they adapt to the new feed, young animals lose weight, their growth is delayed, and they become more susceptible to diseases; this is directly reflected in their productive capacity. For these reasons, many ranchers seek commercially available immune stimulant products that can minimize these negative effects of the feed transition period.
The use of additives of plant origin in the post-weaning or weaning period can minimize productive losses resulting from the adaptation of the animals when their diets are altered. The use of phytogenics (plant-derived products such as essential oils used as additives) is a compelling alternative to bolster animal health; many of these additives stimulate biological functions in animals (Karásková et al., 2015; Volpato et al., 2019), thereby enhancing growth performance. Essential oils in ruminant feed can help improve the use of nutrients, in addition to reducing the impact on the environment (Benchaar et al., 2007); some researchers claim that the addition of phytogenics in the feed of adult goats improves the utilization of nutrients, in addition to stimulating antioxidant responses (Choubey et al., 2015). The putative mechanism involves protection of cells and tissues from excess free radicals produced in physiological or pathological reactions, including cellular respiration. The use of a phytogenic based on thymol, cinnamaldehyde, and carvacrol had potent anti-inflammatory effects, combined with better weight gain in broiler chickens (Reis et al., 2018; Galli et al., 2020). There is a strong correlation between productive performance and inflammatory responses; that is, greater inflammatory responses correlate with lower weight gain. During inflammatory responses, there is high consumption/expenditure of ATP, a molecule no more utilized to produce meat and milk. Supplements with antioxidant and anti-inflammatory properties are therefore potentially desirable in animal feed.
We tested a blend based on carvacrol, thymol, and cinnamaldehyde in micro-encapsulated form. This formulation is important for ruminants because it passes in inert form through the rumen and reaches the intestine to be absorbed, thereby improving bioavailability. It is worth mentioning that the herbal components present in this product have antimicrobial activity. Since antiquity, properties of essential oils have been used; it is now recognized that these have antiviral, antibacterial, antifungal, and anti-inflammatory effects (Acamovic and Brooker, 2005). Among the activities of essential oils, their antimicrobial effects have been demonstrated in a wide variety of microorganisms, including gram-positive and gram-negative bacteria (Benchaar et al, 2008). It is not desirable that the product be released in the rumen, where there are protozoa and bacteria that could be negatively affected by the herbal components. Recently, a study conducted by Chapman et al. (2019) revealed that addition of 2 and 4 mg cinnamaldehyde/kg diet of dairy cows exerted antimicrobial effect in the rumen, but did not alter feed intake, milk yield or milk components. On the other hand, Chapman et al. (2017) revealed that inclusion of 1 and 2 mg cinnamaldehyde/kg diet not exert performance or coccidia control benefits for weaned heifers, suggesting that it is dose dependent. Carvacrol is a phenolic monoterpenoid. It positions itself in bacterial cytoplasmic membranes and increases permeability and fluidity, thereby allowing passive transport of ions through the membrane (Hyldgaard et al., 2012). Lambert et al. (2001) reported that the mixture of carvacrol and thymol caused total inhibition of Pseudomonas aeruginosa and Staphylococcus aureus, mediated by damage caused to the integrity of the membrane, affecting pH homeostasis and the balance of inorganic ions.
Therefore, the hypothesis in the present study was that the use of a phytogenic additive in the diet would improve growth performance, and antioxidant and immune responses in lambs. The objective of the current study was to determine whether supplementation with a micro-encapsulated commercial product (a phytogenic) formulated with carvacrol, thymol, and cinnamaldehyde in lamb feed would improve performance, as well as antioxidant and immune responses.
Section snippets
Phytogenic blend
We used a commercial phytogenic (Enterosan®, Konkreta Feed, Navegantes, SC) containing essential micro-encapsulated oil components. According to the manufacturer, the product contains carvacrol, thymol and cinnamaldehyde. The levels of these components were measured as described below.
We extracted the micro-encapsulated herbal components to quantify the active ingredients. We transferred 0.05 g of the microcapsules to 2-mL vials and added 1 mL of acetonitrile. Samples were homogenized in vortex
Feed intake
Lambs in all three groups consumed more than 95% of the feed (silage and hay) provided daily (control = 96.7%; T500 = 95.3%; and T1000 = 96.5%); and 100% concentrate was consumed by animals of three groups.
Phytogenic analyses and daily intake of carvacrol, thymol and cinnamaldehyde per animal
The micro-encapsulated commercial product contained 21.55 mg carvacrol/g, 18.76 mg thymol/g, and 27.62 mg cinnamaldehyde/g. Based on these results, the levels of each herbal component used in the lamb feed were calculated to contain the following:
T500 (10.77 mg carvacrol/kg, 9.38 mg thymol/kg
Discussion
We observed greater weight gain in T500 and T1000 animals than in Control animals. In another study, Ornaghi et al. (2017) evaluated supplementation with essential oils from cinnamon and cloves and reported greater weight gain in the animals in the treated group than in the Control group. This effect was related to the presence of volatile and odorant compounds in the essential oils that influence ingestion of dry matter, likely due to the improvement in palatability caused by these oils. In
Conclusion
Supplementation with the phytogenic product in lamb feed increases weight gain, likely in association with an anti-inflammatory effect. This supplementation changes the antioxidant profile without changing the levels of free radicals and lipid peroxidation, which we believe to be a consequence of the high doses consumed by the animals. Although there was a difference between treatments in terms of cholesterol levels at one point in the study, we nevertheless conclude that the consumption of
Ethical note
The study was approved by the Committee of Ethics in the Use of Animals of the State University of Santa Catarina (CEUA / UDESC) under number 9502300918.
Authors' contribution
Favaretto J.A., Alba D.F., Bianchi A.E. and Da Silva A.S. contributed to the design and implementation of the research, to the analysis of the results. Wagner R and Vedovatto M. helped in the elaboration of the project and its execution and financing. Marchiori M.S., Marcon H.J. and Zanluchi M. participated in the execution of the experiment and collection of samples and data. Souza C.F., Baldissera MD and Klein B. did the laboratory analysis. All authors discussed the results and contributed
Declaration of Competing Interest
The authors declare no conflict of interest.
Acknowledgements
We thank CAPES, CNPq, and UDESC for financial support. We also thank Cabanha Três Leite that made the animals available and the facility for this study.
References (36)
- et al.
Effects of essential oils on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition in dairy cows fed alfalfa silage or corn silage
J. Dairy. Sci.
(2007) - et al.
A review of plant-derived essential oils in ruminant nutrition and production
Anim. Feed Sci. Technol.
(2008) - et al.
Role of essential oils in food safety: Antimicrobial and antioxidant applications
Grain Oil Sci. Technol.
(2019) - et al.
Weaning distress in dairy calves: effects of alternative weaning procedures
Appl. Anim. Behav. Sci.
(2008) - et al.
Effects of cinnamaldehyde or monensin on performance of weaned Holstein dairy heifers
J. Dairy Sci.
(2017) - et al.
Combination of herbal components (curcumin, carvacrol, thymol, cinnamaldehyde) in broiler chicken feed: Impacts on response parameters, performance, fatty acid profiles, meat quality and control of coccidia and bacteria
Microb. Pathog.
(2020) - et al.
Glutathione S-transferases: the first enzymatic step in mercapturic acid formation
J. Biol. Chem.
(1974) - et al.
Quantification of lipid peroxidation in tissue extracts based on Fe (III) xylenol orange complex formation
Free Radic. Biol. Med.
(1995) - et al.
Effect of carvacrol essential oils on immune response and inflammation-related genes expression in broilers challenged by lipopolysaccharide
Poult. Sci.
(2019) - et al.
Essential oils in the feed of young bulls: Effect on animal performance, digestibility, temperament, feeding behaviour and carcass characteristics
Anim. Feed Sci. Technol.
(2017)
Effects of phytogenic feed additive based on thymol, carvacrol and cinnamic aldehyde on body weight, blood parameters and environmental bacteria in broilers chickens
Microb. Pathog.
Effect of essential oils on meat and fat qualities of feedlot finished young bulls
Meat Sci.
Effects of feed supplementation with clove and rosemary essential oils and protected oils (eugenol, thymol and vanillin) on animal performance, carcass characteristics, digestibility, and ingestive behavior activities for Nellore heifers finished in feedlot
Livestock Sci.
Symposium: carbohydrate methodology, metabolism, and nutritional implications in dairy cattle
J. Dairy Sci.
A theoretically-based model for predicting total digestible nutrient values of forages and concentrates
Anim. Feed Sci. Technol.
Official Methods of Analysis
Biochemistry of plant secondary metabolites and their effects in animals
Proc. Nutr. Soc.
Toxicity of thymol, carvacrol and their respective phenoxyacetic acids in Lactuca sativa and Sorghum bicolor
Ind. Crops Prod.
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