Marigold Meal Research Articles

Marigold and Orange Skin as Egg Yolk Color Promoting Agents

The research was conducted with natural ingredients and orange skin in the diet of laying pullets to determine the suitability as pigmenting agents of egg yolk for laying chicken The influence of tes t ingredients on production characteristics of laying pullets, internal and external quality characteristics of eggs were investigated simultaneously. The experiment covered the proximate analysis of marigold, orange skin meal to determine the nutrient concentrations, xanthophylls contents and 7-12 weeks feeding trail to th th investigate the yolk pigment availability of marigold and orange skin as well as laying performance of birds. Orange skin and marigold were used for comparing the pigmenting ability to normal feed containing xanthophylls. Yolk color was improved significantly as compared to control for addition of marigold and orange skin during 4 , 8 and 12 weeks of supplementation. Inclusion of marigold and orange skin in the th th th diet of laying pullets did not cause deterioration in natural and external quality of eggs and there were no significant variations with respect to body weight, hen day egg production egg weight and feed conversion. There was no mortality of birds during study period. The results indicates the conclusion that between two natural ingredients marigold contain more xanthophylls (156.32mg/kg DM) than orange skin (83.02mg/kg DM) and use of 4% marigold meal in the diet of laying pullets is enough to produce eggs with yolk color score 11.00 close to 30 mg synthetic pigment/kg diet at the 12 week of supplementation, whereas the birds that th received 4% orange skin in the diet were able to produce eggs with yolk color score 5.0. Moreover, use of marigold and orange skin in diet of pullet has no detrimental effect on internal and external quality of egg as well as egg production characteristics. So, marigold can be used more efficiently for egg yolk pigmentation.

A Method for Estimating the Relative Degree of Saponification of Xanthophyll Sources and Feedstuffs

Saponification of xanthophyll esters in various feed sources has been shown to improve pigmentation efficiency in broiler skin and egg yolks. Three trials were conducted to evaluate a rapid liquid chromatography procedure for estimating the relative degree of xanthophyll saponification using samples of yellow corn, corn gluten meal, alfalfa, and 6 commercially available marigold meal concentrates. In each trial, samples were extracted using a modification of the 1984 Association of Official Analytical Chemists hot saponification procedure with and without the addition of KOH. A comparison of the chromatography results was used to estimate percent saponification of the original sample by dividing the nonsaponified extraction values by the saponified extraction values. A comparison of the percent saponified xanthophylls for each product (mg/kg) was: yellow corn, 101; corn gluten meal, 78; alfalfa, 97.9; and marigold concentrates A through F, 99.8, 4.6, 99.0, 95.6, 96.8, and 6.6, respectively. These results indicate that a modification of the 1984 Association of Official Analytical Chemists procedure and liquid column chromatography can be used to quickly verify saponification and can be used to estimate the relative degree of saponification of an unknown xanthophyll source.

The performance of young pigs fed different amounts of marigold ( Calendula officinalis) meal; a pilot study

Hexane-extracted calendula meal was tested in an acceptance trial with eighty 8–13 weeks old crossbred pigs to determine their response to diets containing 0, 2, 6, 10 or 20% calendula meal. Performance parameters included feed intake, daily growth and post mortem histopathological examination of vital organs. Pigs fed a diet containing 2% calendula meal consumed significantly more feed than the ones fed a calendula-free diet. As the calendula meal content of the diet increased, feed intake tended to decrease. Post mortem examination of vital organs showed statistically significant ( P < 0.10) between-treatment differences in heart, kidney, thyroid and pancreas size expressed as percentage of bodyweight. The results of blood and blood serum analyses for haematocrit, haemoglobin, oxygenated haemoglobin, aspartate-aminotransferase, alanine-aminotransferase, lactate dehydrogenase, creatinine and zinc remained within the reference values for young pigs. Although calendula meal showed potential as a ration ingredient for young pigs it is advised not to include more than 10% of it in the diet.

Effects of Enzymatic Treatments of Marigold Flowers on Lutein Isomeric Profiles

Reversed-phase C30 HPLC was applied to study the identity of lutein isomers and to monitor the effects of solids content and elimination of water-soluble substances on the isomeric carotenoid profiles of marigold (Tagetes erecta) samples treated with enzymes. The tentative identity of four lutein isomers present in saponified marigold extracts was confirmed. Enzymatic treatment on a 5% solids slurry produced the marigold meal with the highest all-trans-lutein content [25.1 g/kg dry weight (dw)]. We did not observe variations in the distribution in percentage of lutein isomers due to enzymatic treatment; the elimination of water solubles had a significant but small effect on such variations. The solids content was the principal factor that affected the carotenoid profiles. An analysis of the distribution showed that 15% solids gave the highest all-trans-lutein percentage in treated meals. Interestingly, with 20% solids both the degradation of lutein and the percentage of all-trans-zeaxanthin were the highest. Keywords: Lutein; carotenoids; pigments; marigold; Tagetes

Enzymatic treatment to enhance carotenoid content in dehydrated marigold flower meal.

The effect of enzymatic treatments using a commercial enzyme (Econase-cep at pH 5.0 and 0.1% w/w concentration) at different levels of dehydrated marigold meal (5, 10, 15 and 20% dry weight), to enhance carotenoid extraction, was evaluated. The AOAC method consisting of a simultaneous extraction and saponification by using hexane-ethanol-acetone-toluene (10:6:7:7 v/v) and metanolic KOH (40% w/v) was used to evaluate the carotenoid content. The measurement of carotenoids was also carried out in samples in which the water soluble compounds were previously eliminated (AOAC-H2O). Total carotenoids ranged from 11.4 to 17.4 g/kg and 18 to 24.7 g/kg of control and treated marigold meal, respectively. Highest amount of carotenoids were noted when 5% level of treated meal was used.

Correlation of HPLC and AOAC Methods to Assess the All-trans-Lutein Content in Marigold Flowers

A reversed-phase C30 high-performance liquid chromatography (HPLC) method was used to separate the lutein isomers of marigold flowers. The pigments were extracted from marigold meal by an Association of Official Analytical Chemists (AOAC) modified method. The unsaponified extract showed the presence of several lutein ester isomers and after saponification four major lutein isomers were detected. The relationship between all-trans-lutein, assessed by HPLC, and total xanthophylls, assessed by AOAC, was determined. A regression equation, with a high correlation coefficient (r2=0·972), which allows for the calculation of all-trans-lutein from total xanthophylls content is proposed.

Correlation of HPLC and AOAC Methods to Assess the All-trans-Lutein Content in Marigold Flowers

A reversed-phase C30 high-performance liquid chromatography (HPLC) method was used to separate the lutein isomers of marigold flowers. The pigments were extracted from marigold meal by an Association of Official Analytical Chemists (AOAC) modified method. The unsaponified extract showed the presence of several lutein ester isomers and after saponification four major lutein isomers were detected. The relationship between all-trans-lutein, assessed by HPLC, and total xanthophylls, assessed by AOAC, was determined. A regression equation, with a high correlation coefficient (r2=0·972), which allows for the calculation of all-trans-lutein from total xanthophylls content is proposed.

Effects of Formaldehyde Fumigation of Housing on Carotenoid Pigmentation in Three Breeds of Chickens

Three breeds of chickens (two broiler breeds, A and B, and one layer breed, C) were raised from 1 day to 3 wk of age in either standard housing or housing additionally fumigated with formaldehyde. Groups of chicks from each breed in both housings received either regular poultry starter ration or this ration supplemented with marigold meal. At 3 wk of age, chicks of all three breeds had higher (111, 113, and 115%) mean body weights when raised in fumigated housing. There was no significant effect of supplemental carotenoids on body weight. Chicks of all three breeds raised in fumigated housing had significantly higher plasma carotenoid concentrations when fed either regular or supplemented feed, and, with the exception of Breed A on regular feed, also had significantly increased skin carotenoid values. No significant differences among breeds in plasma or skin carotenoid values were seen in chicks fed regular ration. However, significant differences were seen among groups of chicks fed supplemental carotenoids. The layer breed, C, had the highest percentage increase in carotenoid pigmentation when fed the carotenoid-supplemented diet in fumigated housing.

Modification of Official Method for Analysis of Carotenes and Xanthophylls in Dried Plant Materials and Mixed Feeds

The official method (43.018-43.023) for extraction and separation of carotenes and xanthophylls in dried plant materials and mixed feeds is modified. These pigments are extracted from samples lacking xanthophyll esters by stirring 2 hr, followed by 1-hr cold saponification. The official method recommends 16-hr static extraction preceding the cold saponification. For samples containing xanthophyll esters, we preceded the 20-min hot saponification step by 2 hr stirring. This extra step increased the yield of mono- and dihydroxy pigments, especially from marigold meal. For separation, Merck silica gel 60 plus Hyflo Supercel is substituted for silica gel G in adsorbent I, resulting in 30-min separation. Also, at least 4 separations can be performed using the same adsorbent, and the column is more easily cleaned between separations. Results of the modified method were comparable to those of the official method.

Xanthophyll Availability of Alfalfa Leaf Protein Concentrate (Pro-Xan) for Broilers and Laying Hens

The availability of the xanthophyll of alfalfa leaf protein concentrate (Pro-Xan) was evaluated in two trials with broilers and one trial with laying hens. In the broiler trials, freeze-dried Pro-Xan and Pro-Xan dried in a pilot plant Heil dehydrator at outlet temperatures of 76.7 and 90.6°C. were shown to have xanthophyll availabilities of about 1.7 times that of dehydrated alfalfa and three times that of marigold meal. The xanthophyll availability of dehydrated alfalfa was about half that of pure lutein and twice that of marigold meal. Similar results for xanthophyll availability were found for freeze-dried Pro-Xan, dehydrated alfalfa, and lutein in the laying hen trial.

Effect of Storage and Fatty Acid Esters on the Utilization of Xanthophyll from Marigold Meal by Laying Hens

The feeding of two different harvests of marigold meal on an equivalent analyzed basis to laying hens produced no difference in NEPA numbers, β-carotene equivalents, and fan yolk scores for their eggs. This indicates that there are no structure rearrangements in the xanthophylls which are traceable to the storage of the marigold meal. Marigold meal samples were shown to lose from 21–28% of the xanthophyll content over a 10 month period in a refrigerated environment.Hens fed extracts from saponified marigold meal produced eggs with NEPA numbers 15% higher than did those from hens fed nonhydrolyzed marigold meal. The biological availability of marigold meal was 51% for nonhydrolyzed marigold and 66% for saponified marigold compared to β-apo-8′-carotenoic ethyl ester stabilized beadlets. The digestibilities of marigold and alfalfa meal xanthophylls were 88% and 96%, respectively, when fed to hens at a concentration of 33 mg./kg. diet. The hydrolysis of the 61% natural esterified xanthophyll in marigold meal increased the digestibility to 97%. The data suggest the esterified xanthophylls in marigold meal decreases the bioavailability for egg yolk pigmentation.

Pigmentation of Egg Yolks by Xanthophylls from Corn, Marigold, Alfalfa and Synthetic Sources

Leghorn-type hens in cages and floor pens were used to evaluate pigmenting potential for producing table-egg and intensely colored egg yolks from the xanthophylls in corn gluten meal, alfalfa and marigold meal and for β-apo 8′ carotenal (BAC) and the carotenoic acid ethyl ester (BACE). There were no consistent differences in utilization efficiencies between corn gluten meal, 20% protein dehydrated alfalfa meal, a 40% protein alfalfa concentrate, total xanthophyll extracts of corn or alfalfa or BAC. Xanthophylls in marigold meal, although most concentrated, were least utilizable and BACE was most efficiently utilized. Lutein isolated from alfalfa was not utilized as well at levels up to 22 p.p.m. as the xanthophylls of alfalfa meal or the alfalfa extracts, suggesting the presence of a factor in the latter materials which enhances xanthophyll utilization. Utilization was superior for caged hens vs. floor housed hens.

Use of Heat to Saponify Xanthophyll Esters and Speed Analysis for Carotenoids in Feed Materials: Collaborative Study

Abstract Twelve collaborators analyzed 4 pairs of samples to compare a 20 min hot (56°C) saponification technique with the overnight extraction technique at room temperature in the official first action method for analysis of carotenoids in feed materials, 39.018–39.023, with a view to (a) make the method applicable to products which contain xanthophyll esters and (b) speed up the analysis. In general, the collaborators were in good agreement, with saponification of esters essentially complete after the 20 min heating of samples that contained marigold meal. While the sum of extracted pigments from corn gluten and alfalfa meal tended to be slightly higher with the longer (overnight) contact with the solvent than with the hot extraction, the differences were not statistically significant. Significant differences between hot and cold treatments were absent also in alfalfa pigment fractions, but hot saponification significantly lowered monohydroxy pigments and increased dihydroxy pigments in extracts from corn gluten. The results showed good precision with either extraction technique. The method, modified to include hot saponification as an alternative step, is now applicable to all dried feed materials, including those which contain xanthophyll esters (e.g., marigold meal); the modification has been incorporated into the official first action method, 39.018–39.023.

The Use of Beta-Apo-8′-Carotenoic Acid Ethyl Ester as a Standard for Determination of Xanthophyll Utilization from Natural Sources by Broilers

A synthetic pigment, beta-apo-8′-carotenoic acid ethyl ester, was evaluated as a standard from which xanthophyll utilization by broilers could be determined for various feed ingredients. The ingredients tested were yellow corn, alfalfa meal, corn gluten meal, kenaf meal, marigold meal and canthaxanthin. Xanthophyll utilization from each ingredient was determined by comparison with standard curves developed for beta-apo-8′-carotenoic acid ethyl ester at levels of 5, 10, 20, 30 and 40 grams per ton of feed. Pigmentation measurement of both skin and shanks was accomplished using visual scoring systems and a reflectance colorimeter.It was concluded that beta-apo-8′-carotenoic acid ethyl ester could be used effectively as a standard in xanthophyll utilization studies. Both visual scoring and instrumental determinations of pigmentation appeared to be efficient in determining the xanthophyll utilization values.

Oxycarotenoids in Poultry Pigmentation: 2. Broiler Studies

IN addition to egg yolk color (Marusich and Bauernfeind, 1970), dietary oxycarotenoids in the rations of chickens are needed (1) to adequately pigment broilers and fryers, (2) to provide shank and skin pigmentation in replacement pullets at the start of the laying period and (3) to endow day-old chicks with acceptable yellow color. Yellow corn, corn gluten meal and dehydrated alfalfa meal have served as the primary sources of oxycarotenoids (Day and Williams, 1958; Blessin et al., 1958; Ratcliff et al., 1959, 1962; Waldroup et al., 1960; Dua et al., 1967; Combs and Nott, 1967; Wilkinson and Barbee, 1968; Kuzmicky et al., 1968; and Livingston et al., 1969). In addition, several other feed additives have been shown to be effective including algae meal (Perdue and Smith, 1962; Smith and Perdue, 1966); and marigold meal (Brambila et al., 1963). In the present studies, the chemically synthesized oxycarotenoids, 4,4′ diketo-β-carotene (canthaxanthin) and…