β-Carotene oxidation products have newly discovered bioactivity in plants and animals. Synthetic fully oxidized β-carotene (OxBC) has application in supporting livestock health, with potential human applications. The safety of synthetic OxBC has been evaluated. An Ames test showed weak-to-moderate mutagenicity in only one cell line at high concentrations. A mouse micronucleus assay established a non-toxic dose of 1800 mg/kg body weight, and no bone marrow micronuclei were induced. Plant sources of β-carotene inevitably contain varying levels of natural OxBC. Vegetable powders and dried forages can be especially rich. Intakes of natural OxBC for humans and livestock alike have been estimated. The exposure range for humans (1-22 mg/serving) is comparable to the safe intake of β-carotene (<15 mg/d). In livestock, OxBC in alfalfa can contribute ~550-850 mg/head/d for dairy cattle but in forage-deficient poultry feed much less (~1 ppm). Livestock intake of supplemental synthetic OxBC is comparable to OxBC potentially available from traditional plant sources. Human intake of synthetic OxBC in meat from livestock fed OxBC is similar to a single serving of food made with carrot powder. It is concluded that consumption of synthetic OxBC at levels comparable to natural OxBC is safe for humans and animals.

In animals carotenoids show biological activity unrelated to vitamin A that has been considered to arise directly from the behavior of the parent compound, particularly as an antioxidant. However, the very property that confers antioxidant activity on some carotenoids in plants also confers susceptibility to oxidative transformation. As an alternative, it has been suggested that carotenoid oxidative breakdown or metabolic products could be the actual agents of activity in animals. However, an important and neglected aspect of the behavior of the highly unsaturated carotenoids is their potential to undergo addition of oxygen to form copolymers. Recently we reported that spontaneous oxidation of ß-carotene transforms it into a product dominated by ß-carotene-oxygen copolymers. We now report that the polymeric product is biologically active. Results suggest an overall ability to prime innate immune function to more rapidly respond to subsequent microbial challenges. An underlying structural resemblance to sporopollenin, found in the outer shell of spores and pollen, may allow the polymer to modulate innate immune responses through interactions with the pattern recognition receptor system. Oxygen copolymer formation appears common to all carotenoids, is anticipated to be widespread, and the products may contribute to the health benefits of carotenoid-rich fruits and vegetables.

We reported previously that the spontaneous oxidation of β-carotene and other carotenoids proceeds predominantly by formation of carotenoid-oxygen copolymers and that β-carotene copolymers exhibit immunological activity, including priming innate immune function and limiting inflammatory processes. Oxidative loss of carotenoids in fruits and vegetables occurs during processing. Here we report evidence for the occurrence of associated analogous copolymer compounds. Geronic acid, an indirect, low molecular weight marker of β-carotene oxidation at ∼2% of β-carotene copolymers, is found to occur in common fresh or dried foods, including carrots, tomatoes, sweet potatoes, paprika, rosehips, seaweeds, and alfalfa, at levels encompassing an approximately thousand-fold range, from low ng/g in fresh foods to μg/g in dried foods. Copolymers isolated from several dried foods reach mg/g levels: comparable to initial carotenoid levels. In vivo biological activity of supplemental β-carotene copolymers has been previously documented at μg/g levels, suggesting that some foods could have related activity.

Oxidized β-carotene (OxBC), a phytochemical that occurs naturally in plants, is formed by the spontaneous reaction of β-carotene with ambient oxygen. Synthetic OxBC, obtained by full oxidation of β-carotene with air, shows considerable promise as an in-feed antimicrobial alternative additive that enhances health and performance in livestock. OxBC is predominantly composed of β-carotene-oxygen copolymers that have beneficial immune-modulating effects that occur within the innate immune system by priming it to face microbial challenges and by mitigating the inflammatory response. OxBC does not have any direct anti-bacterial activity. Further, unlike traditional immune stimulants, OxBC modulates but does not stimulate and utilize the animal’s energy stores unless directly stress-challenged. These immune effects occur by mechanisms distinct from the provitamin A or antioxidant pathways commonly proposed as explanations for β-carotene’s actions. Trials in poultry, swine, and dairy cows with low parts-per-million in-feed OxBC supplementation have shown performance benefits over and above those of feeds containing regular vitamin and mineral premixes. Through its ability to enhance immune function, health, and performance, OxBC has demonstrated utility not only as a viable alternative to in-feed antimicrobials but also in its ability to provide tangible health and performance benefits in applications where antimicrobial usage is precluded.

β-carotene spontaneously copolymerizes with molecular oxygen to form a β-carotene–oxygen copolymer compound (“copolymer”) as the main product, together with small amounts of many apocarotenoids. Both the addition and scission products are interpreted as being formed during progression through successive free radical β-carotene–oxygen adduct intermediates. The product mixture from full oxidation of β-carotene, lacking both vitamin A and β-carotene, has immunological activities, some of which are derived from the copolymer. However, the copolymer’s chemical makeup is unknown. A chemical breakdown study shows the compound to be moderately stable but nevertheless the latent source of many small apocarotenoids. GC–MS analysis with mass-spectral library matching identified a minimum of 45 structures, while more than 90 others remain unassigned. Newly identified products include various small keto carboxylic acids and dicarboxylic acids, several of which are central metabolic intermediates. Also present are glyoxal and methyl glyoxal dialdehydes, recently reported as β-carotene metabolites in plants. Although both compounds at higher concentrations are known to be toxic, at low concentration, methyl glyoxal has been reported to be potentially capable of activating an immune response against microbial infection. In plants, advantage is taken of the electrophilic reactivity of specific apocarotenoids derived from β-carotene oxidation to activate protective defenses. Given the copolymer occurs naturally and is a major product of non-enzymatic β-carotene oxidation in stored plants, by partially sequestering apocarotenoid metabolites, the copolymer may serve to limit potential toxicity and maintain low cellular apocarotenoid concentrations for signaling purposes. In animals, the copolymer may serve as a systemic source of apocarotenoids.

Fully oxidized β-carotene (OxBC) containing β-carotene-oxygen copolymers is proposed as an alternative to antimicrobial growth promoters. Two trials were conducted to determine the efficacy of OxBC in enhancing growth and feed intake (FI) in male and female Ross × Ross 308 broilers in Ontario, Canada, and in Ross 308 male broilers in the United Kingdom. In the first trial, 0, 1, 2, or 5 ppm OxBC were added to diets in a 20% cornstarch premix, whereas in the second trial, 0, 2, or 5 ppm OxBC were added in a 1% cornstarch or 1% corncob grits premix. In trial 1, 2, and 5 ppm OxBC improved bird final body weights (BW) compared with the unsupplemented, nonmedicated (no bacitracin methylene disalicylate included), negative control birds after 39 d of feeding under commercial conditions (P < 0.05). All levels of OxBC improved feed conversion (FCR) during the finisher period (P < 0.05), whereas 2 and 5 ppm OxBC enhanced FCR relative to the negative control group during the full production cycle (P < 0.05). Average daily FI was not affected by OxBC, whereas 2 and 5 ppm OxBC increased broiler average daily gain (ADG) (P < 0.05). Oxidized β-carotene did not affect bird mortality. The optimal OxBC dose was 2 ppm under the conditions used. In trial 2, 2 or 5 ppm OxBC on cornstarch and 5 ppm OxBC on corncob grits improved ADG, BW, and FI when fed for 35 d, as compared with the negative, nonmedicated control (P < 0.05). Feed conversion was not improved in the OxBC groups compared with the control group (P > 0.05). There were no differences among the 3 OxBC groups (P > 0.05). When birds were fed 2 ppm OxBC on corncob grits, the overall ADG, BW, and FI were lower than the respective control values (P < 0.001). Overall bird mortality was higher than expected for all groups, including the control group, but no dose effect was evident. Cornstarch was the preferred carrier for OxBC, and 2 ppm OxBC was the optimal dose under the conditions used.