Dietary effects of lutein-fortified chlorella on milk components of Holstein cows
- Jin-Young Jeon†2,
- Keun-Kyu Park†1,
- Kyung-Woo Lee1,
- Seung-Wan Jang2,
- Byung-Hern Moon3 and
- Byoung-Ki An1Email author
© The Author(s) 2016
Received: 16 March 2016
Accepted: 17 June 2016
Published: 28 June 2016
This study was conducted to investigate the dietary effect of conventional or lutein-fortified chlorella on milk production and lutein incorporation in milk. Fifteen Holstein cows in mid-lactation were used in a 3 × 3 Latin square design each with a 21-day period. Cows were top-dressed daily with 30 g of conventional or lutein-fortified chlorella for 3 weeks. Cows without chlorella served as the control. The feed intake and milk yield were not affected by dietary treatments. The concentrations of milk protein and solids non-fat in groups fed diets containing both conventional and lutein-fortified chlorella were significantly higher than those of the control group (P < 0.001). There was no significant difference in content of milk fat among groups. The levels of plasma glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, interferon-gamma and interleukin-2 were not influenced by the dietary treatments. Lutein content in milk was significantly increased in groups fed lutein-fortified chlorella as compared with those of conventional chlorella and control, respectively (P < 0.01). These results imply that conventional and lutein-fortified chlorella has positive effects on milk components and the use of lutein-fortified chlorella in a dairy diet is effective in the production of milk enriched with lutein.
Lutein is a natural fat soluble xanthophyll carotenoid occurring as a pigment in some plants and algae. Food sources of natural lutein include green and yellow vegetables and fruits, egg yolks and breast milk (Sommerburg et al. 1998). Several carotenoids, such as lutein and zeaxanthin, occur in significant concentrations in human macula and retina and they are often referred to as macular pigments (Sommerburg et al. 1998). Particularly, lutein, has been known to reduce aged-macular degeneration and cataracts (Olmedilla et al. 2003) and has a strong antioxidant capacity (Sindhu et al. 2010).
Chlorella, a genus of unicellular green algae, is a good source of lutein. Compared with higher plants, chlorella has an advantage of being able to be cultivated in bioreactors on a large scale as a continuous and reliable source of product (Jeon et al. 2012). In previous studies, we found that the lutein content of the chicken eggs was greatly increased after feeding of chlorella powder to commercial laying hens (Jeon et al. 2012). Recently, there have been increasing interests in dietary supplementation of carotenoids and vitamin E to enhance the composition of the fat soluble fractions, particularly lutein, in milk and dairy products (Calderon et al. 2007). In addition, lutein in its role as an antioxidant directly influences nutritional quality of the dairy products (Antone et al. 2012). Therefore, we evaluated the dietary effects of conventional chlorella and lutein-fortified chlorella on milk components and lutein content in milk of dairy cows.
Animals, diets and management
Proximal composition and lutein concentration of conventional and lutein-fortified chlorella
Crude protein (%)
Ether extract (%)
Crude fiber (%)
Crude ash (%)
Formula and chemical composition of experimental diet
Corn gluten feed
Cotton seed meal
Vitamin and mineral mixa
Calculated valuesb (%)
Total digestible nutrients (%)
Sampling and measurements
Cows were milked two times daily in their stall and milk yield was recorded every day. Milk samples were collected on weekly basis and concentrations of fat, protein, solids non-fat (SNF) and lactose were analyzed using a Fossomatic-605 infrared analyzer (Foss Electric, Hillerod, Denmark). An aliquot of milk was prepared separately for further analyses.
Lutein contents in milk were also determined according to the method of Schlatterer and Breithaupt (2006), with some modification. In brief, an aliquot of samples was placed in a round-bottom flask with 45 mL of ternary solvent mixture (light petroleum/ethyl acetate/methanol; 1:1:1, v/v/v). Two milliliters of distilled water was added to the flask to facilitate separation. The separation was involved two immiscible liquid phases, the upper layer phase was recovered. After vacuum evaporation (50 mBar, 30 °C, 10 min), the extract including fatty residues was transferred to the volumetric flask with TMBE/methanol (1:1, v/v) to a total volume of 10 mL. The extracts were filtered through a No. 6789 0.45 µm filter membrane (Whatman International Ltd., Maidstone, England) and assayed using high-performance liquid chromatography (HPLC; Beckman Coulter Inc., CA, USA).
At the end of each period, blood samples were taken from the jugular vein in heparinized vacutainer tubes. The heparinized tubes were immediately chilled on ice box after sampling, centrifuged at 2000×g for 15 min and stored at −60 °C until analysis. The levels of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and total cholesterol (TC) were measured according to the colorimetric method using a modular system biochemical analyzer (Hitachi Ltd., Tokyo, Japan). Bovine interferon-gamma (IFN-γ) was measured using the Bovine IFN-γ ELISA Kit (Cusabio Biotech Co. Ltd., Wuhan, China) as suggested by the manufacturer. The concentration of plasma interleukin-2 (IL-2) was also measured using the Bovine IL-2 ELISA Kit (Cusabio Biotech Co. Ltd.). The samples and standard curves were measured in triplicate.
Results and discussion
Dietary effects of conventional or lutein fortified chlorella powder on yield and components of milks in dairy cows
Feed intake (kg/head/day) (on DM basis)
Milk yield (kg/head/day)
Milk components (g/kg)
The positive effect of cows fed chlorella on milk protein and SNF may be mediated through the lutein derived from chlorella and/or changes in ruminal fermentation. Xu et al. (2014) reported that supplementation of lutein in the diet can improve milk components, such as fat and lactose, and reduce SCC in lactating cows. The ruminal fermentation by microbes enables growing ruminants to utilize cellulose from chlorella very efficiently (Chowdhury et al. 1995). Feeding algae, including chlorella and Scenedesmus, increases fiber digestibility and feed efficiency in growing calves (Chowdhury et al. 1995). Milk yield and/or composition can be greatly influenced by changes in ruminal fermentation via microbial activity (Li et al. 2012). But there are little data available concerning the effect of dietary chlorella on ruminal fermentation in dairy cows. The possibility of changed ruminal fermentation should not be precluded and further study is required to clarify dietary effects of chlorella on ruminal micro-organisms. Milk components are economically important to both milk producers and processors and nutritionally important to consumers. The present results suggest that the conventional or lutein-fortified chlorella is effective in improving milk protein and SNF when applied to dairy cow diets.
Dietary effects of conventional or lutein fortified chlorella powder on blood profiles in dairy cows
Some studies suggest a variety of physiological and pharmacological effects of chlorella powder and extracts of chlorella, including lipid metabolism, immunomodulatory and antibacterial activity (Hasegawa et al. 1997; Shibata et al. 2001). An et al. (2008) reported significantly increased levels of serum IFN-γ in chlorella fed mice as compared with the non-fed control, especially under the condition of energy-protein malnutrition. In contrast, Queiroz et al. (2002) found that chlorella extract administration did not affect levels of all cytokines measured in normal/non infected mice. In this study, the plasma IFN-γ and IL-2 in groups fed diets with lutein-fortified chlorella were slightly higher than those of other groups, but not significantly (Table 4). Therefore, it is likely that immune-stimulating effect by dietary chlorella does not always occur because differences in animal species, dosage levels, feeding frequency and health status.
Dietary effects of conventional or lutein fortified chlorella powder on lutein concentration in dairy cows
Lutein (µg/L milk)
Dietary chlorella has positive effects on milk components in Holstein cows and the use of conventional or lutein fortified chlorella is effective for the production of lutein fortified milk.
JYJ and KKP carried out the experimental work and wrote the manuscript. SWJ, KWL and BHM helped in the data analysis. BKA provided funding and designed the experiment. All authors read and approved the final manuscript.
This paper was supported by the KU Research Professor Program of Konkuk University.
The authors declare that they have no competing interests.
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