Canthaxanthin Mitigates Cardiovascular Senescence in Vitro and in Vivo.

BACKGROUND: The number of older people in the world is increasing year by year; studies have shown that more than 90% of cardiovascular disease occurs in the older people population, indicating that aging is one of the major risks involved in the development of cardiovascular disease. Therefore, retarding the development of cardiac aging is an important strategy to prevent aging-related cardiovascular diseases. METHODS: In the current study, we examined the anti-cardiovascular aging potential of canthaxanthin in vitro and in vivo experiments. For this, a model of cardiomyocyte senescence induced by D-galactose was established, which was used to investigate the canthaxanthin's effect on cardiac premature aging. RESULTS: We found that canthaxanthin obviously mitigated the cardiomyocyte senescence in vitro. Further mechanistic studies revealed that canthaxanthin seems to alleviate cardiomyocyte senescence by regulating the autophagy process. Furthermore, the effects of canthaxanthin on cardiovascular senescence were further evaluated. We also observed that canthaxanthin mitigated cardiac aging and fibrosis in the aged mice model. CONCLUSIONS: To sum up, the current work showed that canthaxanthin could obviously alleviate cardiac premature aging, indicating that canthaxanthin could be used as a biologically active molecule for the treatment of cardiac aging and fibrosis.

Dietary supplementation of spirulina and canthaxanthin boosts laying performance, lipid profile in blood and egg yolk, hatchability, and semen quality of chickens.

This study examined the impacts of dietary spirulina (SP) and canthaxanthin (CX) supplementation to laying hens on reproductive performance and cock's semen quality evaluation. Three hundred and sixty of indigenous Sinai chickens (300 hens + 60 cocks), 48-weeks-old, were randomly divided into five treatments; a control and four supplemented (0.6 g SP mg/kg, 0.8 g SP mg/kg, 6 mg CX/kg diet, and 8 mg CX/kg diet) with six replications of 10 females and 1 male per treatment for laying trial and 1 male for semen evaluation. The animals were fed with experimental diets for 12 weeks. Supplementing 6 mg CX/kg diet led to an increase in the final body weight and egg production traits, while 8 mg CX/kg diet resulted in an improving feed conversion ratio. Supplementation of 0.6 g SP mg/kg or 6.0 mg CX/kg diet resulted in the highest egg production. The heaviest egg weight and egg mass were induced by 0.8 g SP mg/kg. Eggs laid from hens fed 0.6 g SP/kg diet had a higher concentration of cholesterol than those fed 0.8 g SP/kg and those fed 6.0 and 8.0 mg CX/kg. Cocks fed 6 mg CX/kg and those fed 0.8 g SP/kg diet had the greatest sperm concentration. The supplementation of 0.8 g SP mg/kg diet or both 6 and 8 mg CX/kg diet enhanced hatchability. So, dietary spirulina (0.8 mg/kg) and canthaxanthin (6 mg/kg) supplementation to Sinai laying hens and cocks could be used to improve productive and reproductive and performance.

Canthaxanthin Attenuates the Vascular Aging or Endothelial Cell Senescence by Inhibiting Inflammation and Oxidative Stress in Mice.

BACKGROUND: Vascular endothelial dysfunction is an early phenotype of aging-related vascular dysfunction. Delaying vascular aging and preventing cardiovascular disease are major public health problems that urgently need to be solved. Scientists have studied various drugs to prevent the occurrence and progress of cardiovascular disease, but progress has been slow. Here, the antisenescence and anti-endothelial damage of canthaxanthin (CX, which is an active molecule from food) has been studied. METHODS: This study was performed by adding CX to a model of cell senescence and oxidative damage induced by hydrogen peroxide. Cellular senescence markers (e.g., p16, p21, and p53) and oxidative damage markers (e.g., reactive oxygen species, nitric oxide, malondialdehyde, superoxide dismutase) were evaluated by the enzyme-linked immunosorbent assay, laser scanning confocal microscopy, and Western blotting. RESULTS: We found that CX downregulated the expression level of senescence-associated molecules, and significantly reduced the oxidative damage of vascular endothelial cells. These observations showed that CX effectively alleviated the senescence of vascular endothelial cells. Furthermore, CX treatment reduced the expression levels of interleukin-6 (IL-6), tumor necrosis factor alpha, and IL-1ß. Finally, in vivo, CX significantly alleviated vascular senescence. CONCLUSIONS: The current study shows that CX has potential application value for treating vascular aging or endothelial cell senescence.

The effects of canthaxanthin microencapsulation on yolk color and canthaxanthin deposition in egg yolk of laying hens.

Canthaxanthin is widely used as a feed additive to improve skin and yolk color in poultry. It is insoluble in water and sensitive to oxidation, so commercial canthaxanthin is often microencapsulated with wall materials to improve its solubility and stability. The objective of this study was to evaluate the effects of canthaxanthin microencapsulation on yolk color and canthaxanthin deposition in egg yolk of laying hens. A total of 288 Hyline Brown laying hens (48 wk of age) were allocated to 4 groups with 6 replicates of 12 hens each, and fed a basal diet or the basal diet supplemented with 5 mg/kg canthaxanthin microencapsulated with modified starch (CMMS), gelatin (CMG), and sodium lignosulfonate (CMSL), respectively. Canthaxanthin supplementation did not affect laying performance of hens, but improved (P < 0.05) yolk color of fresh, fried, boiled, and stored (4 and 25°C) eggs. The improvement of yolk color of fresh eggs was greatest in the CMSL group and least in the CMG group (P < 0.05). Both CMMS and CMSL resulted in higher (P < 0.05) yolk canthaxanthin concentration than CMG. The CMSL resulted in higher (P < 0.05) yolk color score of fried eggs than CMMS and CMG and higher (P < 0.05) yolk color score of boiled eggs than CMG, but no difference was observed in stored eggs among three canthaxanthin groups. In conclusion, CMMS and CMSL were more effective in yolk pigmentation than CMG, and CMSL was slightly better than CMMS.

Dietary supplementation with canthaxanthin and 25-hydroxycholecalciferol on the incubation performance and fertility of European quail breeders.

This study assessed the effects of combined supplementation with canthaxanthin (Cx) and 25-hydroxycholecalciferol (25-OH-D3) on incubation performance, fertility, and chick quality in European quail breeders. A total of 240 birds were distributed in a completely randomized design with 5 diets and 8 replicates. The animals were fed a basal diet containing 50 µg of vitamin D3 or the basal diet supplemented with 3 ppm Cx and 34.5 µg 25-OH-D3, 6 ppm Cx, and 69 µg 25-OH-D3, 9 ppm Cx and 103.5 µg 25-OH-D3, or 12 ppm Cx and 138 µg 25-OH-D3. Incubation performance was analyzed in 2 periods (32 and 38 wk). Breeders aged 32 wk produced eggs with higher hatchability (P = 0.024), hatchability of fertile eggs (P = 0.026) and lower initial plus mid embryonic mortality (P = 0.021), whereas 38-week-old breeders generated chicks with a higher length at hatching (P < 0.001) and lower final plus pipped embryonic mortality (P = 0.021). In both age groups, Cx + 25-OH-D3 levels had a quadratic effect on egg fertility (P < 0.001), hatchability of total (P < 0.001), and fertile eggs (P < 0.001). The fertility and the number of sperm cells in the perivitelline membrane was analyzed in two periods (26 and 40 wk). A quadratic effect of diet and days after mating on both parameters (P < 0.05) was observed. Eggs from supplementing breeders showed a high fertility (P < 0.001) and sperm cell counts (P < 0.001) for up to 7 and 3 d after mating, respectively, then the control group. Moreover, the supplementation of quail breeder diets with 6 ppm Cx + 69 µg 25-OH-D3 enhances sperm cell longevity in sperm storage tubules, hatchability of total and fertile eggs, fertility, and chick quality, especially in older quail's breeders and reduces embryonic mortality.

Culture optimization to enhance carotenoid production of a selected purple nonsulfur bacterium and its activity against acute hepatopancreatic necrosis disease-causing Vibrio parahaemolyticus.

Purple nonsulfur bacteria (PNSB) were investigated for their carotenoid production and anti-vibrio activity against acute hepatopancreatic necrosis disease (AHPND)-causing Vibrio parahaemolyticus. To test carotenoid production, selected strains were cultivated in basic isolation medium (BIM), glutamate acetate medium, G5 medium and artificial acetic acid wastewater (AAW) medium. From 144 PNSB, Rhodopseudomonas palustris KTSSG46 was selected to produce carotenoids under microaerobic light conditions in BIM. When the culture medium was optimized, strain KTSSG46 grown in BIM modified with l-glutamate at 1 g/L more effectively inhibited AHPND-causing V. parahaemolyticus strains than standard BIM with 1 g/L (NH4 )2 SO4 . BIM was further modified with 1.23 g/L MgSO4 ·7H2 O and carotenoid production increased 40.22%. Carotenoid production at day 2 by strain KTSSG46 grown in BIM modified with l-glutamate at 1 and 1.23 g/L MgSO4 ·7H2 O was the same as production in BIM modified with monosodium glutamate (MSG). Culture supernatants from all BIM formulations showed similar activity against the resistant AHPND strain SR2. Based on high-performance liquid chromatography, carotenoids of strain KTSSG46 might be canthaxanthin. Grown in BIM modified with MSG, strain KTSSG46 could produce inexpensive carotenoids and release anti-vibrio compounds that, applied as shrimp feed additive, would prevent AHPND strains.

Effect of a commercial product containing canthaxanthin for in ovo feeding to broiler embryos on hatchability, chick quality, oxidation status, and performance.

In ovo feeding has been indicated to improve hatchability, newly hatched chick quality, and broiler performance. The aim of this study was to investigate the effect of in ovo feeding of a commercial canthaxanthin product (CCX) containing lignosulphonate, corn starch, canthaxanthin, dextrin (yellow), and ethoxyquin through assessing incubation results, newly hatched quality and oxidation status and broiler performance at 1 to 14 d of age. A total of 780 egg were distributed in a randomized complete block design with 5 treatments (levels of CCX: 0.0, 0.35, 0.45, 0.55, and 0.65 mg/0.5 mL of sterilized and distilled water) and 156 eggs per treatment. The blocking factor was setters. At 17.5 d of embryo development, in ovo injected treatments were applied, using a manual needle. The in ovo feeding of CCX resulted in lower hatching rates (P < 0.05) and a longer hatching window (P < 0.05) as compared with noninjected CCX treatment. The CCX injection did not affect the bursa and spleen percentage of newly hatched chick (P > 0.05). In addition, a higher percentage of chicks with poor physical quality score (<71.0 points) was obtained among the chicks from eggs injected with 0.55 and 0.65 mg of CCX (P < 0.05). There were higher total proteins and catalase activity in the livers of the chicks injected with CCX. Broiler chicks in the control group (0.0 mg of CCX) presented higher BW and BW gain during 1 to 7 and 7 to 14 d of after hatch (P < 0.05). The viability (%) of chicks at 1 to 14 d of after hatch decreased with inoculation greater than 0.45 mg of CCX in ovo (P < 0.05). Although the CCX shown an improvement in oxidation status of chicks, the hatchability and performance of broilers decreased. We concluded that a commercial CCX is not recommended for injection in ovo, and furthers studies should carried out to elucidate the use of pure canthaxanthin.

Dietary supplementation with canthaxanthin and 25-hydroxycholecalciferol has beneficial effects on bone and oxidative metabolism in European quail breeders.

This study aimed to investigate the effect of supplementation with canthaxanthin (Cx) and 25-hydroxycholecalciferol (25-OH-D3) on the production performance, egg quality, bone mineral content, blood biochemical parameters, and antioxidant status of European quail breeders. Two hundred and forty quail breeders were distributed in a completely randomized design with 5 diets and 8 replicates of 4 females and 2 males were used. All quail breeders received one of 5 diets: basal diet (containing 2,000 IU vitamin D3) or the same diet supplemented with 3 ppm Cx and 34.5 µg 25-OH-D3, 6 ppm Cx and 69 µg 25-OH-D3, 9 ppm Cx and 103.5 µg 25-OH-D3, or 12 ppm Cx and 138 µg 25-OH-D3. Production performance and internal and external egg quality parameters were not influenced by diet. Eggshell dry weight decreased linearly with increasing supplementation levels, and eggshell ash and calcium content increased quadratically. Plasma phosphorus, calcium, and ionic calcium levels in females and plasma ionic calcium levels in males showed a positive quadratic response to dietary supplementation. Femoral and tibiotarsal dry weight and calcium content were influenced by diet. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity in the liver of males and females and in the serum of females showed a positive quadratic relationship with Cx and 25-OH-D3 levels, whereas the malonaldehyde concentration showed a negative quadratic relationship. DPPH scavenging activity in the serum of male quail increased linearly with supplementation. There was a positive quadratic effect on superoxide dismutase gene expression and a positive linear effect on glutathione peroxidase 7 gene expression, suggesting that dietary enrichment with Cx and 25-OH-D3 might help protect spermatozoa against oxidative damage. The dietary supplement was pro-oxidative at high concentrations (above 9 ppm Cx). The results indicate that diets with adequate levels of Cx and 25-OH-D3 have a beneficial effect on calcium and phosphorus metabolism as well as on the antioxidant defense system. We recommend supplementing European quail breeders in the laying period with 6 ppm Cx and 69 µg 25-OH-D3.

Activation of TMEM16A by natural product canthaxanthin promotes gastrointestinal contraction.

Transmembrane 16A (TMEM16A), also known as anoctamin 1, is the molecular basis of the calcium-activated chloride channels. TMEM16A is present in interstitial cells of Cajal, which are the pacemaker cells that control smooth muscle contraction. TMEM16A is implicated in gastrointestinal disorders. Activation of TMEM16A is believed to promote the gastrointestinal muscle contraction. Here, we report a highly efficient, nontoxic, and selective activator of TMEM16A, canthaxanthin (CX). The study using molecular docking and site-directed mutation revealed that CX-specific binging site in TMEM16A is K769. CX was also found to promote the contraction of smooth muscle cells in gastrointestinal tract through activation of TMEM16A channels, which provides an excellent basis for development of CX as a chemical tool and potential therapeutic for gastrointestinal dysfunction.

Canthaxanthin protects human sperm parameters during cryopreservation.

Different antioxidants have been introduced to reduce oxidative stress during the cryopreservation. The main goal of this study was to evaluate the effects of canthaxanthin on human sperm parameters during the freeze-thaw process. This study was performed on 25 normozoospermic semen samples dividing into five groups including 0, 0.1, 1, 10, and 25 µM of canthaxanthin. The prepared spermatozoa were cryopreserved by rapid freezing technique. Sperm motility, viability (eosin-nigrosin), morphology (Papanicolaou), acrosome reaction (double staining), DNA denaturation (acridine orange), chromatin packaging (aniline blue and toluidine blue), and DNA fragmentation (sperm chromatin dispersion test) were evaluated before freezing and after thawing. All sperm parameters after thawing significantly were decreased compared to before freezing. Twenty-five micromolar canthaxanthin could significantly improve the progressive and total motility, viability, normal morphology, chromatin packaging, acrosome integrity and DNA denaturation and fragmentation. Ten micromolar canthaxanthin significantly improved total motility, viability, normal morphology, chromatin packaging, acrosome integrity and DNA denaturation and fragmentation. Whereas, in 1 µM group, there were significant differences only in improvement of acrosome integrity, chromatin packaging (toluidine blue) and DNA denaturation and fragmentation. But, in 0.1 µM group, there were no significant differences in any of measured parameters. It seems that canthaxanthin ameliorates detrimental effects of cryopreservation on human sperm parameters.

Effects of astaxanthin and canthaxanthin on oxidative stress biomarkers in rainbow trout.

Farmed trout are commonly fed carotenoid-enriched diets during the finishing period to acquire typical red-to-pink flesh color in salmonid muscle. The aim of this study was to examine the effects of two xanthophylls, astaxanthin (Ax) or canthaxanthin (Cx), administered individually or in combination, on oxidative stress biomarkers in kidney and liver of rainbow trout. Specimens were fed Ax (75 mg/kg) or Cx (25 mg/kg) individually or in combination in the diets for 8 weeks. Changes in concentration of oxidative stress biomarkers, including total glutathione, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione S-transferase, were recorded in Ax- and Cx-dosed trout. These two carotenoids, predominantly Cx, initiated enzymatic responses in rainbow trout. It is noteworthy that lipid peroxidation processes were not apparent in all Ax or Cx-dosed trout. Further, both combined xanthophylls did not exert significant synergistic effects in liver and kidney. Biomarker responses were generally altered in both tissues through the 4 and 8 weeks suggesting that different time-dependent mechanisms led to enhanced antioxidant defense in Ax and/or Cx-fed trout. Data demonstrated that these two xanthophylls did not exert detrimental effects on rainbow trout.

The dietary supplementation of canthaxanthin in combination with 25OHD3 results in reproductive, performance, and progeny quality gains in broiler breeders.

The objective of this study was to evaluate the effects of the dietary supplementation of canthaxanthin and cholecalciferol (25OHD3) to broiler breeders and their progenies on their performance. Eighty 25-wk-old Cobb 500 broiler breeders were distributed according to a completely randomized design in 2 experimental treatments, with 10 replicates of 4 birds each. The treatments consisted of the supplementation or not of a corn-soybean meal diet with canthaxanthin + 25OHD3 to broiler breeders fed from 25 to 62 wk of age. Egg production, fertility, hatchability, hatchability of fertile eggs, embryonic mortality, and egg specific gravity were evaluated. Breeders were inseminated at 35, 45, and 62 wk of age, the eggs were incubated, and the performance of the progenies was evaluated. From the progeny of each breeder age, 300 male chicks were distributed according to a completely randomized design in a 2 × 2 factorial arrangement (chicks from breeders supplemented or not with canthaxanthin + 25OHD3 and chicks supplemented or not with canthaxanthin + 25OHD3), totaling 4 treatments with 5 replicates of 15 birds each. Canthaxanthin + 25OHD3 were supplied to the chicks until 21 D of age. The combination of canthaxanthin and 25OHD3, containing 69 mg of 25OHD3 and 6 g of canthaxanthin, was supplemented at 1 kg/t of feed. Breeders supplemented with canthaxanthin + 25OHD3 showed higher egg production, total hatchability, hatchability of fertile eggs, and lower early embryo mortality compared with those fed the control diet. Broilers from breeders fed canthaxanthin + 25OHD3 and supplemented with this additive up to 21 D of age presented a better feed conversion ratio and higher carcass and breast yields than those derived from nonsupplemented breeders.

Dietary canthaxanthin reduces xanthophyll uptake and red coloration in adult red-legged partridges.

Carotenoids give color to conspicuous animal signals that are often the product of sexual selection. Knowledge of the mechanisms involved in carotenoid-based signaling is critical to understanding how these traits evolve. However, these mechanisms remain only partially understood. Carotenoids are usually viewed as scarce dietary antioxidants whose allocation to ornaments may trade off against health. This trade-off would ensure its reliability as a signal of individual quality. In the case of red (keto)carotenoids, the literature suggests that some species may show constraints in their uptake. Canthaxanthin is one of the most common ketocarotenoids in red ornaments of animals. It is often commercially used as a dietary supplement to obtain redder birds (e.g. poultry). We increased the dietary canthaxanthin levels in captive red-legged partridges (Alectoris rufa). This species shows red non-feathered parts mostly pigmented by another common ketocarotenoid: astaxanthin. We studied the impact on the uptake of carotenoids and vitamins and, finally, on coloration. We also tested the potential protective effect of canthaxanthin when exposing birds to a free radical generator (diquat). Canthaxanthin did not apparently protect birds from oxidative stress, but interfered with the absorption of yellow carotenoids (lutein and zeaxanthin). Zeaxanthin is a precursor of astaxanthin in enzymatic pathways, and their levels in tissues and eggs were lower in canthaxanthin-supplied birds. This led to lower astaxanthin levels in ornaments and paler coloration. As far as we know, this is the first report of a carotenoid supplementation decreasing animal coloration. The results have implications for understanding carotenoid-based signaling evolution, but also for improving husbandry/experimental procedures.

Effect of dietary canthaxanthin and iodine on the production performance and egg quality of laying hens.

In this study, we aimed to evaluate the effect of canthaxanthin (CX) and iodine (I) on the production of laying hens, on counteracting debilitation of the vitelline membrane, and on inhibiting Salmonella growth in eggs stored at 30°C. Three hundred hens were reared in cages. Birds were divided into six feeding groups (10 hens × 5 repetitions) that were administered 0, 3 or 6 ppm of CX and 1 or 10 ppm of I with their diets. Laying rate, egg weights, and feed conversion ratios were controlled. The quality of fresh eggs was assessed in wks 25-26, 48-50 and 62-63 of hens lives. An additional batch of eggs was incubated at the temperature of 30°C, and egg quality changes were monitored on days 3, 6 and 9 of storage. Additionally, eggs collected from four experimental groups of hens whose diets had been iodated with 1 or 10 ppm of I and supplemented with 0 or 6 ppm of CX were infected under laboratory conditions with Salmonella, and incubated for 5 and 10 d. The laying rate, egg weights, and feed conversion ratio were significantly improved. Dietary inclusion of CX contributed to a higher resistance of the vitelline membrane of egg yolks, but only for fresh eggs. Vitelline membrane degradation during egg storage at 30°C was significantly counteracted by dietary inclusion of I at a dose of 10 ppm. The same I dose resulted in the complete inhibition of Salmonella growth until day 10 of incubation, but exclusively for eggs collected from 40-week-old hens. Dietary supplementation with 10 ppm of I was found to impart high level of resistance to the vitelline membrane against the growth of Salmonella in case of eggs stored at 30°C; therefore, I was found to be more beneficial by ensuring longer preservation than that of CX. However, dietary supplementation with CX was found to increase the resistance of vitelline membrane in fresh eggs.

Anti-Skin Cancer Activities of Apostichopus japonicus Extracts from Low-Temperature Ultrasonification Process.

OBJECTIVES: This work aimed to enhance anti-skin cancer activities of Apostichopus japonicus, spiky sea cucumber, through ultrasonification extraction process at low temperature. METHODS: Dried Apostichopus japonicus was extracted with an ultrasonification process at 50°C and 95 kHz for two hours (UE), and anti-skin cancer activities of the extract from the UE were also compared with those from conventional extraction processes using hot water (WE) or 70% ethanol at 80°C (EE) for 12 hours. RESULTS: The amount of canthaxanthin in the UE was higher than that in the WE or EE, and its cytotoxicity against human keratinocytes was less than the others. The extract from the UE showed 93.5% inhibition against human malignant cell growth, which was also higher than those from both WE and EE. The extract from the UE demonstrated the ability of inhibiting both cancer cell proliferation and metastasis by downregulating the skin tumor-promoting genes such as Bcl-2, STAT3, and MMP-9. CONCLUSIONS: The ultrasonification process was proved to be effective especially in extracting heat-sensitive marine biomass, A. japonicus having higher amounts of canthaxanthin and better anti-skin cancer activities, possibly due to less destruction and high elution of bioactive substances under low temperature extraction condition.

Effect of broiler breeders fed with corn or sorghum diet and canthaxanthin supplementation on production and reproductive performance.

This study was designed to evaluate the effect of corn or sorghum diet and canthaxanthin (CX) supplementation on performance in broiler breeders. A total of 440 females with similar body weight (BW) (3.71 ± 0.14 kg) and 60 roosters were placed in an open-sided house with 20 pens, randomly distributed in a factorial arrangement (2 × 2). There were 4 diets of 2 ingredients; corn (CO) or sorghum (SO) and 2 levels of CX; 6 mg/kg (CX) and 0 mg/kg (NCX) totaling 5 replicate pens of 22 females and 3 males each, from 42 to 65 wk, divided in 2 periods (from wk 42 to 53 and wk 54 to 65). Birds' BW was measured every 28 d and mortality rate was calculated at the end of trial. Egg production (%), egg specific gravity (g/cm3), egg weight (g), yolk weight (%), albumen weight (%), eggshell weight (%) and yolk colorimetric score were measured weekly. Incubation parameters were recorded in 12 incubations to evaluate hatching eggs, hatching (%), hatchability (%), fertility (%), weight of the chicks born and their quality. The BW, mortality, percentage of yolk and albumen weight, fertility and some incubation parameters were not affected (P > 0.05) by diets used. An increase in the egg production, hatching eggs, chicks born and first quality chick by hen at the second period were observed in CX breeder's diets (P = 0.0066; P = 0.0060; P = 0.0368; P = 0.0326). Egg specific gravity and eggshell weight were improved at the first period by SO+CX diet (P = 0.0138; P = 0.0209) and the same effect to egg weight, but at the second period (P = 0.0251). The CX was well absorbed from the diet and effectively transferred to the egg yolk, thereby increasing egg yolk pigmentation in the both periods (P < 0.0001). The CX supplementation in broiler breeder diets improved the productive and reproductive performance (laying% and hatchable eggs) at the second period, also to the both periods improved the egg yolk pigmentation.

Effect of maternal canthaxanthin and 25-hydroxycholecalciferol supplementation on the performance of ducklings under two different vitamin regimens.

This study investigated the effects of maternal canthaxanthin (CX, 6 mg/kg) and 25-hydroxycholecalciferol (25-OH-D3 , 0.069 mg/kg) supplementation on the performance of Cherry Valley ducklings under two different vitamin regimens. A total of 780 duck breeder females and 156 males were randomly allotted to two diets with or without the addition of the mixture of CX and 25-OH-D3 (CX+25-OH-D3 ) for 32 weeks. Ducklings (males and females separately) hatched from eggs laid at 24 weeks of the duck breeder trial were fed with a NRC vitamin regimen, and ducklings (males and females separately) hatched from eggs laid at 32 weeks of the duck breeder trial were fed with a HIGH vitamin regimen (had higher levels of all vitamins except biotin than NRC vitamin regimen), for 14 days. The results showed that, maternal CX+25-OH-D3 supplementation increased the shank pigmentation for 7-days post hatch in ducklings under a NRC vitamin regimen, and for 14-days post hatch in ducklings under a HIGH vitamin regimen. Growth performance, antioxidant status and serum phosphorus of ducklings under a NRC vitamin regimen were increased by maternal CX+25-OH-D3 supplementation; however, these positive effects were not observed in ducklings under a HIGH vitamin regimen. Males revealed increased growth performance in ducklings under both NRC and HIGH vitamin regimens. Sexual differences in shank pigmentation, antioxidant status, tibia strength and serum phosphorus were not consistent as they were dependent on maternal CX+25-OH-D3 status or dietary vitamin regimens. Data suggest that maternal CX+25-OH-D3 supplementation is important for starter ducklings under a NRC vitamin regimen, but not HIGH vitamin regimen.

The Effect of Canthaxanthin on the Quality of Frozen Ram Spermatozoa.

The addition of antioxidants to semen cryopreservation extenders has been employed for combating oxidative damage. This work aimed to evaluate the addition of carotenoid canthaxanthin to a cryopreservation extender of ram semen. Three breeder rams were used and, after semen collection, with 48-hour intervals between collection, the samples were included in the pool formation (n = 6). The experimental groups comprised 0 (control), 0.1, 1, 10, and 25 µM of canthaxanthin. After thawing (37°C/30 s) and incubation at 37°C for 2 hours, semen aliquots from each group were evaluated for sperm kinetics (CASA), the integrity of the plasma and acrosomal membranes (iPAM), intracellular reactive oxygen species (ROS) production, and lipid peroxidation (LPO) by flow cytometry associated with the image. The control group and canthaxanthin 1 µM after incubation at 37°C for 2 hours showed increases of curvilinear velocity and amplitude of lateral head displacement with decreases of linearity, straightness, and wobble (p < 0.05), which were not observed for the canthaxanthin 10 and 25 µM. The supplementation of a Tris-egg yolk extender with canthaxanthin had no effect on the iPAM, intracellular ROS production in viable spermatozoa, or LPO. In conclusion, supplementation with 10 and 25 µM of canthaxanthin in a Tris-egg yolk extender used for ram semen cryopreservation is able to protect ovine sperm from kinetic changes after incubation at 37°C for 2 hours post-thawing.

The effects of canthaxanthin on porcine oocyte maturation and embryo development in vitro after parthenogenetic activation and somatic cell nuclear transfer.

The objective of this study was to examine the effects of canthaxanthin (Cx) treatment during in vitro maturation (IVM) of porcine oocytes on embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT), on intracellular glutathione (GSH) and reactive oxygen species (ROS) levels in mature oocytes, and on gene expression in both PA- and SCNT-derived blastocysts. To determine the optimal effective concentration of Cx, porcine oocytes were cultured in IVM medium supplemented with various concentrations (0, 20, 40 and 80 µM) of Cx for 22 hr. Compared to other groups, supplementation with 40 µM Cx significantly improved blastocyst formation rates after PA (p < .05), but no significant differences were observed among groups in total blastocyst cell numbers. Subsequently, oocytes were cultured in IVM medium supplemented with or without 40 µM Cx. Oocytes treated with 40 µM Cx showed significantly increased cleavage and blastocyst formation rates after SCNT compared to the control group (p < .05). Moreover, significantly increased intracellular GSH and reduced ROS levels were observed in the Cx-treated group (p < .05). In addition, both PA- and SCNT-derived blastocysts from the 40 µM Cx-treated group showed significantly increased mRNA expression of Bcl2 and Oct4 and decreased Caspase3 expression level (p < .05), when compared with the control group. PA-derived blastocysts from the 40 µM Cx-treated group also exhibited significantly decreased expression of Bax (p < .05). Our results demonstrated that treatment with 40 µM Cx during IVM improves the developmental competence of PA and SCNT embryos. Improvement of embryo development by Cx is most likely due to increased intracellular GSH synthesis, which reduces ROS levels in oocytes, and it may also positively regulate apoptosis- and development-related genes.

In Vitro Inhibition of Human UDP-Glucuronosyl-Transferase (UGT) Isoforms by Astaxanthin, ß-Cryptoxanthin, Canthaxanthin, Lutein, and Zeaxanthin: Prediction of in Vivo Dietary Supplement-Drug Interactions.

Despite the widespread use of the five major xanthophylls astaxanthin, ß-cryptoxanthin, canthaxanthin, lutein, and zeaxanthin as dietary supplements, there have been no studies regarding their inhibitory effects on hepatic UDP-glucuronosyltransferases (UGTs). Here, we evaluated the inhibitory potential of these xanthophylls on the seven major human hepatic UGTs (UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A9, UGT2B7 and UGT2B15) in vitro by LC-MS/MS using specific marker reactions in human liver microsomes (except UGT2B15) or recombinant supersomes (UGT2B15). We also predicted potential dietary supplement-drug interactions for ß-cryptoxanthin via UGT1A1 inhibition. We demonstrated that astaxanthin and zeaxanthin showed no apparent inhibition, while the remaining xanthophylls showed only weak inhibitory effects on the seven UGTs. ß-Cryptoxanthin mildly inhibited UGT1A1, UGT1A3, and UGT1A4, with IC50 values of 18.8 ± 2.07, 28.3 ± 4.40 and 34.9 ± 5.98 µM, respectively. Canthaxanthin weakly inhibited UGT1A1 and UGT1A3, with IC50 values of 38.5 ± 4.65 and 41.2 ± 3.14 µM, respectively; and lutein inhibited UGT1A1 and UGT1A4, with IC50 values of 45.5 ± 4.01 and 28.7 ± 3.79 µM, respectively. Among the tested xanthophyll-UGT pairs, ß-cryptoxanthin showed the strongest competitive inhibition of UGT1A1 (Ki, 12.2 ± 0.985 µM). In addition, we predicted the risk of UGT1A1 inhibition in vivo using the reported maximum plasma concentration after oral administration of ß-cryptoxanthin in humans. Our data suggests that these xanthophylls are unlikely to cause dietary supplement-drug interactions mediated by inhibition of the hepatic UGTs. These findings provide useful information for the safe clinical use of the tested xanthophylls.