Comparison of the Dietary Supplementation of Lactobacillus plantarum, and Fermented and Non-Fermented Artemisia Annua on the Performance, Egg Quality, Serum Cholesterol, and Eggyolk-Oxidative Stability During Storage in Laying Hens

ABSTRACT

Artemisia annua L. is a widely distributed medicinal plant and well-known for treating malaria due to the artemisinin content. We previously found enhanced antioxidant and antibacterial activities of Lactobacillus plantarum-fermented A. annua dried leaves in vitro. The present study compared the effects of the dietary supplementation of L. plantarum, fermented (FA) or non-fermented (NFA) A. annua on laying performance, egg quality, serum cholesterol, and egg yolk oxidative stability during storage in 40-weeks-old Hy-Line Brown layers. In total, 180 layers were randomly allocated into four treatments for 6 weeks: basal diet (CON), basal diet + 0.5% L. plantarum only (LO), basal diet + 0.5% NFA, and basal diet + 0.5% FA. Each treatment comprised five replicates with nine birds each. Egg weight of NFA and FA groups were significantly higher as compared with the CON and LO groups (p<0.01). The FA group displayed higher Haugh unit (HU) compared with the NFA group (p<0.05). Eggshell color of the FA group was significantly increased compared with the other groups (p<0.01). There was no significant difference in triglyceride, total cholesterol, HDL-cholesterol, and VLDL+LDL cholesterol among the different groups. During egg storage, the HU of FA groups was significantly increased as compared with the CON group after 3 weeks storage (p<0.05). The malondialdehyde (MDA) content in the stored eggs was significantly lowered by feeding of FA as compared with the CON and LO groups (p<0.05). Altogether, the fermented A. annua displayed positive effects in promoting egg quality of layers.

Keywords:
Artemisia annua L; egg quality; Lactobacillus plantarum; laying hens; malondialdehyde

INTRODUCTION

There has been an increasing interest in using phytogenic feed additives (PFA) in animal feed during the last two decades (Mohammadi Gheisar & Kim, 2018). To date, many studies have reported antimicrobial, antioxidant, anti-inflammatory, and growth-promoting effects of PFA in animals (Kim et al., 2010; Abdel-Wareth & Lohakare, 2014; Qin & Hou, 2017; Abou-Elkhair et al., 2018; Mohammadi Gheisar & Kim, 2018). Phytogenic sources contain abundant phytochemicals, which have been mainly classified into three categories as carotenoids, isothiocyanates, and polyphenols, and most polyphenols are recognized as having beneficial functions (Fraser, 2009).

A traditional medicinal herb, Artemisia has been used to treat a variety of diseases for a long time. The genus Artemisia comprises over 500 species and it is mainly distributed in the temperate zones of Europe, Asia, and North America. A. annua, also known as sweet wormwood or annual mug wort, is one of the most famous Artemisia species applied to treat several diseases, including malaria, due to its high artemisinin content (Bora & Sharma, 2011). At present, more than 600 secondary metabolites have been identified and classified as terpenoids, flavonoids, coumarins, caffeoylquinic acids, sterols, and acetylenes (Brown, 2010; Abad et al., 2012). The pharmacological effects of A. annua have already been well-studied, but its functional effects as feed additives are not well assessed so far. A few studies have investigated the effects of Artemisia supplementation in broiler diets. Dried A. annua leaves were reported to increase the feed conversion ratio and to reduce intestinal Clostridium perfringens counts in broilers (Engberg et al., 2012). The dietary supplementation of dried A. annua leaves meal lowered lipid oxidation in the breast and thigh muscles of broilers (Cherian et al., 2013). In addition, dried A. annua leaves also displayed anticoccidial effects in free-range broilers (Brisibe et al., 2008; de Almeida et al., 2012). Enzyme-treated A. annua has potential to improve growth performance, antioxidant capacity, tolerance to heat stress, and to alleviate the intestinal inflammatory response of broilers (Wan et al., 2016; Song et al., 2017). A. annua has also exhibited promising economic potential as a feed additive for broilers even before the ban of conventional coccidiostats in EU (Bosselmann & Gylling, 2013).

Probiotics are also well-studied feed additives, with reported benefits on laying performance, egg quality, and immune response of laying hens (Zhang et al., 2012; Forte et al., 2016; Abd et al., 2017; Mazanko et al., 2018). Lactobacillus plantarum is one of the most common probiotic strains, and possesses strong antioxidant and antimicrobial activities (Tsai et al., 2012; Niu et al., 2018). The dietary supplementation of L. plantarum metabolites increased egg production, reduced plasma and yolk cholesterol, and improved fecal volatile fatty acids content (Loh et al., 2014) in layers. Plant materials may be used as probiotic carriers by fermentation, and, in turn, fermented plants with selected autochthonous probiotics may improve its functional properties (Hossain et al., 2012; Lokaewmanee et al., 2012; Peres et al., 2012; Di cagno et al., 2013; Zhao et al., 2013). Phenolic and flavonoid compounds and their antioxidant capacity in herbal teas were increased by lactic acid fermentation (Ibrahim et al., 2014). Enhanced antioxidant and antibacterial capacities were also reported in a fungi-fermented medicinal plant (Bletilla striata) (Dong et al., 2015). The supplementation of Lactobacillus-fermented Artemisia princeps has been reported to improve the growth performance, meat lipid stability, and gut health of broilers (Kim et al., 2012a).

A suitable probiotic strain is essential to achieve synergetic effects in phytogenic fermentation. We had previously observed enhanced antioxidant and antibacterial activities of L. plantarum-fermented A. annua L. (Lee et al., 2017). To the best of our knowledge, very few studies regarding the dietary effects of fermented A. annua L. in laying hens have been reported. Therefore, we compared the effects of the L. plantarum, fermented and non-fermented A. annua L., in layer diets on laying performance, egg quality, serum cholesterol, and egg yolk oxidative stability during storage.

MATERIALS AND METHODS

Experimental diets, birds, and manage-ment

In the study, the experimental feed additives were prepared as follows. L. plantarum SK3494 was cultured in MRS (deMan, Rogosa and Sharpe, Difco, USA) medium at 37ºC for 24 h to achieve 1.0×10⁹ CFU/mL cells and used as a probiotic feed additive (LO). A volume of 10% (w/v) A. annua L. dried leaves powder in 10-fold diluted MRS medium was used as phytogenic feed additive (NFA). The fermented A. annua L. (FA) was prepared by inoculating 2% (v/v) of L. plantarum SK3494 in 10-fold diluted MRS containing 10% (w/v) A. annua L. dried leaves powder and cultured at 37°C for 24 h to achieve 1.0×10⁹ CFU/mL cells. Finally, 0.5% (w/v) of each additive was evenly mixed with the basal diet (Table 1) in a feed blender and used as experimental diets. The supplemented diets were prepared freshly each 2 weeks to maintain the shelf life. The basal diet not supplemented with feed additives was used as a control feed (CON). The feeding experiment was conducted for 6 weeks.

Table 1
Feedstuffs and calculated chemical composition of the basal diet.

Ingredients	Composition (%)
Corn		50.00
Tallow		0.90
Corn dried distiller’s grains with solubles (DDGS)		21.24
Soybean meal		7.42
Rapeseed meal		5.00
Sesame seed oil meal		2.00
Feather meal		1.50
Syn. Lysine-sulfate		0.38
Syn. Methionine (liq.)		0.108
Syn. Threonine		0.026
Limestone		10.20
Mono dicalcium phosphate (MDCP)		0.59
Salt		0.20
Sodium bicarbonate		0.10
Vitamin premix1		0.11
Mineral premix2		0.18
Choline-chloride (50%)		0.049
Total		100.00
Calculated nutritional composition
Crude protein, %		17
Crude fat, %		5.3
Crude fiber, %		3.7
Ca, %		4.0
Available P, %		0.27
Lys, %		0.83
TSAA4, %		0.72
TMEn3, kcal/kg		2,800

• ¹
  Vitamin premix provided followings per kg of diet: vitamin A, 40,000 IU; vitamin D3, 8,000 IU; vitamin E, 10 IU; vitamin K3, 4 mg; vitamin B1, 4 mg; vitamin B2, 12 mg; vitamin B6, 6 mg; vitamin B12, 0.02 mg; pantothenic acid, 20 mg; folic acid, 2 mg; nicotinic, 60 mg.
• ²
  Mineral premix provided followings per kg of diet: Fe, 30 mg; Zn, 25 mg; Mn, 20 mg; Co, 0.15 mg; Cu, 5 mg; Se, 0.1 mg.
• ³
  TMEn: true metabolizable energy corrected for nitrogen.
• ⁴
  TSAA: Total sulfur amino acids.

The procedures with animals were performed according to the guidelines of the animal policy of the Institutional Animal Care and Use Committee at Konkuk University, Seoul, South Korea.

One hundred eighty 40-week-old Hy-Line Brown layers were randomly assigned to four treatments with five replicates of nine birds each. Hens were housed in a two-tier iron cage (length, 90 cm; width, 90 cm; space allowance, 735 cm²) with nine birds each.

Hens were housed at 22 ± 3°C and 76.4 ± 15.7% humidity, with 16 h of light per day, and were fed the experimental diets and water ad libitum.

Determination of laying performance

All the eggs laid, including intact and broken eggs, were collected daily to determine hen-day egg production ratio (EPR). Average egg weight (AEW) was determined using only intact eggs to divide the egg number. Daily egg mass was calculated by multiplying EPR with AEW on the same day. The feed intake was recorded weekly and expressed as g/day/hen. The feed conversion ratio (FCR) was determined as grams of feed intake per grams of egg mass.

Determination of egg quality

Egg quality was measured weekly in three eggs per replicate. The eggs were individually weighed and exposed to a breaking force in an eggshell strength tester (FHK, Fugihira, Ltd, Japan). After breaking, egg contents were poured into a glass plate to measure albumen height. Haugh unit (HU) was calculated using the records of albumen height (H) and egg weight (W) following the formula [HU = 100 Log (H + 7.57 - 1.7 W^0.37)] as described in Haugh (1937). Egg yolk color was determined by comparing the color with egg yolk color fan (Yolk color fan, Roche, Switzerland). Eggshell color was determined by comparing the color with an eggshell color fan (Eggshell color fan, Samyang, Korea). Eggshell thickness was measured at the center of eggshell fragments using a micrometer (Digimatic Micrometer, Series 293?330, Mitutoyo, Japan).

Determination of serum cholesterol

At the end of the experiment, eight laying hens per treatment were selected for blood sampling. Birds were sacrificed using carbon dioxide and then cardiac blood was collected. The collected blood was stored at 4ºC for 1 day in EDTA-treated blood collection tubes. Serum was collected by centrifugation at 1,500 g for 10 min and stored at -20°C until use. Serum total cholesterol, total triglyceride, and high-density lipoprotein (HDL) cholesterol levels were determined in a biochemical analyzer (HITACHI 7600, Japan), using the corresponding diagnostic kits (Youngdong Medical Corporation, Korea) according to the manufacturer’s direction. VLDL (very low-density lipoprotein) +LDL (low-density lipoprotein)-cholesterol was calculated by subtracting HDL-cholesterol from the total cholesterol as described earlier (Nishizawa & Fudamoto, 1995).

Determination of egg freshness and lipid oxidation during storage

Haugh units and egg yolk oxidative stability were used to determine egg storage quality. At the end of the experiment (6^th week), four intact eggs per replicate (20 eggs/treatment) were randomly collected and stored at 18°C for 4 weeks to determine HU change (Haugh, 1937). Additional 5 intact eggs per replicate (25 eggs/treatment) were stored at room temperature (15 ± 5°C) for 4 weeks, then they were broken and the egg yolks were stored for 2 more weeks (total 6 weeks). Egg yolk oxidative stability was evaluated by thiobarbituric acid reactive substances (TBARS) assay with some modifications (Botsoglou et al., 1994). Egg yolk samples (1.5 g) were homogenized in hexane with 5% (v/v) aqueous trichloroacetic acid (TCA) and 0.8% (v/v) butylated hydroxytoluene, and then the solution was centrifuged at 1,500g for 5 min. A 2.5-mL aliquot of the bottom layer was removed and mixed with 1.5 mL of 0.8% 2-thiobarbituric acid. After incubation at 70°C for 30 min, absorbance was measured at a range of 400-650 nm using a spectrophotometer (Shimadzu, Model UV-1601, Tokyo, Japan) by producing a third-derivative spectrum. Malondialdehyde (MDA) concentration in the analyzed samples was directly quantified by referring a standard calibration curve using tetraethoxypropane (an MDA precursor).

Statistical analysis

All data obtained in this study were subjected to one-way analysis of variance (ANOVA) using the general linear model (GLM) procedure of SAS statistical software (SAS Institute, 2002). Cage was used as the experimental unit for laying performance data, whereas individual birds as experimental unit for serum cholesterol data. The egg was an experimental unit for egg quality data and storage quality data. Dietary treatment was a fixed factor in all statistical models and the LSMEANS procedure was used to calculate mean values. Significant differences among treatments were determined using Duncan’s multiple range test at the level of p<0.05. The data were presented as mean ± standard deviation (SD).

RESULTS

Laying performance

The effects of the dietary supplementation of the probiotic L. plantarum (LO), fermented (FA), and non-fermented A. annuaL (NFA) on laying performance were compared. There were no significant differences (p>0.05) in performance-related parameters, such as egg production rate, egg mass, feed intake, and FCR with the administration of different feed additives (Table 2). However, egg weight of the groups fed NFA and FA groups was significantly higher compared with the CON and LO groups (p<0.01).

Table 2
Effects of the supplementation of the evaluated feed additives on laying performance.

Item	Treatment1				p-value
	CON	LO	NFA	FA
Egg production, %	85.89 ± 6.35	87.60 ± 8.06	85.50 ± 5.97	86.27 ± 6.92	0.66
Egg weight, g	63.18 ± 1.69b	62.89 ± 1.37b	64.55 ± 2.16a	64.19 ± 1.48a	<0.01
Egg mass, g/hen/d	54.33 ± 5.02	55.08 ± 5.18	55.14 ± 3.39	55.40 ± 4.67	0.83
Feed intake, g/hen/d	113.05 ± 9.77	116.28 ± 6.29	116.35 ± 6.27	115.97 ± 5.48	0.22
FCR2, g feed/g egg	2.08 ± 0.23	2.12 ± 0.19	2.11 ± 0.11	2.12 ± 0.23	0.86

• 1CON: basal diet; LO: basal diet + 0.5% L. plantarum; NFA: basal diet + 0.5% non-fermented A. annua L.; FA: basal diet + 0.5% fermented A. annua L.
• 2FCR: feed conversion ratio, measure of an animal’s efficiency in converting feed mass into increases of the desired output.
• a,bDifferent superscripts (a, b) in the same row indicate significantly (p<0.05) different values in mean score (Mean ± SD) among treatment groups.

Egg quality

The effects of the dietary treatments on the egg quality of laying hens are shown in Table 3. Haugh unit a of the NFA group was significantly lower compared with the CON and FA groups (p=0.03). Eggshell color was significantly increased in the FA group compared to the other groups (p<0.01). No significant difference was found in egg yolk color, eggshell breaking strength, and eggshell thickness with the supplementation of different additives. The FA diet was likely to show a trend on increasing the eggshell breaking strength (p=0.08).

Table 3
Effects of the supplementation of the evaluated feed additives on egg quality.

Item	Treatment1				p-value
	CON	LO	NFA	FA
Haugh unit	93.07 ± 6.48a	92.95 ± 7.67ab	90.69 ± 8.32b	94.01 ± 7.35a	0.03
Eggshell color	10.75 ± 1.13b	10.98 ± 0.97b	11.03 ± 1.18b	11.36 ± 0.90a	<0.01
Egg yolk color	7.17 ± 0.66	7.25 ± 0.73	7.19 ± 0.75	7.22 ± 0.72	0.89
Eggshell breaking strength, kg/cm2	3.20 ± 0.79	3.42 ± 0.79	3.22 ± 0.95	3.45 ± 0.69	0.08
Eggshell thickness, mm	0.36 ± 0.03	0.36 ± 0.02	0.36 ± 0.03	0.37 ± 0.03	0.25

• 1CON: basal diet; LO: basal diet + 0.5% L. plantarum; NFA: basal diet + 0.5% Non-fermented A. annua L.; FA: basal diet + 0.5% fermented A. annua L.
• a,bDifferent superscripts (a, b) in the same row indicate significantly (p<0.05) different values in mean score (Mean ± SD) among treatment groups.

Serum cholesterol

The effects of the dietary treatments on serum cholesterol of laying hens are shown in Table 4. There were no significant differences in triglyceride, total cholesterol, HDL-cholesterol and VLDL+LDL-cholesterol among the different diet groups.

Table 4
Effects of the supplementation of the evaluated feed additives on serum cholesterol (mg/dL).

Item	Treatment1				p-value
	CON	LO	NFA	FA
Triglycerides	1,174.63 ± 419.73	1,557.38 ± 192.98	1,128.25 ± 471.41	1,351.13 ± 434.05	0.14
Total cholesterol	133.63 ± 31.08	159.50 ± 12.92	136.75 ± 32.15	148.88 ± 30.51	0.25
HDL-cholesterol	10.22 ± 3.05	11.70 ± 2.19	12.05 ± 3.57	10.36 ± 2.37	0.48
VLDL+LDL-cholesterol2	123.40 ± 28.41	147.80 ± 11.67	124.70 ± 29.36	138.52 ± 28.49	0.20

• ¹
  CON: basal diet; LO: basal diet + 0.5% L. plantarum; NFA: basal diet + 0.5% Non-fermented A. annua L.; FA: basal diet + 0.5% fermented A. annua L.
• ²
  VLDL+LDL-cholesterol: value calculated by subtracting HDL-cholesterol from the total cholesterol.
• ^a,bDifferent superscripts (a, b) in the same row indicate significantly (p<0.05) different values in mean score (Mean ± SD) among treatment groups.

Egg freshness and lipid oxidation during storage

The effects of the dietary treatments on Haugh units and egg yolk peroxidation during storage are shown in Table 5. Significantly higher HU was only observed in 3-week stored eggs of hens fed FA compared with the CON group (p=0.02) (Table 5).The addition of feed additives did not significantly affect egg yolk MDA concentration compared with the CON group after storage for 4 weeks, but lower MDA values were found in the FA group after storage for 6 weeks (p=0.04).

Table 5
Effects of the supplementation of the evaluated feed additives on Haugh unit and egg yolk oxidative stability during storage.

Period	Treatment1				p-value
	CON	LO	NFA	FA
Haugh unit (1 week)	88.77 ± 4.89	85.39 ± 7.60	85.12 ± 10.44	84.28 ± 4.55	0.27
Haugh unit (2 weeks)	69.88 ± 11.91	73.30 ± 10.69	73.10 ± 7.36	72.78 ± 10.23	0.69
Haugh unit (3 weeks)	60.21 ± 6.47b	62.38 ± 7.50ab	64.28 ± 6.91ab	66.78 ± 3.93a	0.02
Haugh unit (4 weeks)	59.25 ± 8.67	61.24 ± 4.28	61.24 ± 5.22	60.49 ± 6.31	0.79
Haugh unit (Overall)	69.81 ± 14.44	71.47 ± 12.58	71.41 ± 12.05	71.69 ± 10.91	0.79
MDA2 (4 weeks) (μg/g yolk)	0.0122 ± 0.0044	0.0119 ± 0.0040	0.0127 ± 0.0020	0.0123 ± 0.0019	0.98
MDA3 (4+2 weeks) (μg/g yolk)	0.0687 ± 0.0285a	0.0641 ± 0.0247a	0.0419 ± 0.0143ab	0.0313 ± 0.0034b	0.04

• ¹
  CON: basal diet; LO: basal diet + 0.5% L. plantarum; NFA: basal diet + 0.5% Non-fermented A. annua L.; FA: basal diet + 0.5% fermented A. annua L.
• ²
  MDA: Malondialdehyde concentration after storage for 4 weeks.
• ³
  MDA: Malondialdehyde concentration after storage for 4 weeks and additional 2 weeks with open-contamination condition without eggshell.
• ^a,bDifferent superscripts (a, b) in the same row indicate significantly (p<0.05) different values in mean score (Mean ± SD) among treatment groups.

DISCUSSION

Fermentation is a common method to produce biological resources with enhanced beneficial properties (Ng et al., 2011). The present study compared the effects of LO, NFA, and FA supplementation in layer diets. No significant differences in laying performance and egg quality were found by LO addition compared with the CON group, which is consistent with a previous study (Forte et al., 2016). The dietary inclusion of NFA and FA significantly increased the egg weight. This finding agrees with those observed by using fennel and cumin seed to increase egg weight (Aydin et al., 2008; Yalcin et al., 2009; Khan et al., 2013; Abou-Elkhair et al., 2018). Conversely, a high inclusion of A. annua leaves did not significantly increase egg weight, but dramatically decreased FCR (Baghban-Kanani et al., 2018). The inclusion of Artemisia capillaris was reported to increase egg production of laying hens (Kim et al., 2010). Positive effects on body weight gain and feed efficiency were also reported by supplementing Lactobacillus-fermented Artemisia princeps for broilers (Kim et al., 2012a). There are many factors that could lead to the difference in the mentioned results viz., dietary level, processing conditions, surveying purpose, and some others. However, the clear reason regarding the mechanism has not been found yet.

Haugh unit is a very important measure of internal egg quality (Monira et al., 2003). The lower egg HU in NFA group compared with the FA and CON groups is inconsistent with the findings in Baghban-Kanani et al. (2018), which reported inclusion of A. annua leaves did not significantly influence HU. We speculate that some original compounds in A. annua may affect albumen synthesis, and these compounds may be converted into others during fermentation. It was earlier reported that fermented herbs (Artemisia capillaris and Acanthopanax senticosus) increased polyunsaturated fatty acid content in the Longissimus dorsi muscle of pigs compared with non-fermented herbs (Lei et al., 2018). The brown coloration of the eggshell is considered as an important eggshell quality parameter and has a positive impact on consumer preference (Samiullah et al., 2015). The supplementation of phytochemicals has been reported to increase eggshell biliverdin (one eggshell pigment) content (Butler & McGraw, 2013). Thus, the enhanced eggshell coloration in FA diet may be caused by more produced active compounds after fermentation.

For serum cholesterol, dietary supplementation of 1% and 2% Artemisia vulgaris L. leaf powder significantly increased HDL-cholesterol and decreased LDL-cholesterol in broilers (Kim et al., 2012b). In contrast, no impact on reducing serum cholesterol was reported by feeding with 1% Artemisia sieberi leaf extract (Khalaji et al., 2011). Artemisia spp. are well known for their pharmacological, antioxidant, anti-inflammation, and antibacterial activities (Choi et al., 2013; van der Kooy & Sullivan, 2013; Kim et al., 2015). In our study, 0.5% of fermented and non-fermented A. annua L. were used and showed little effects on serum cholesterol. The inconsistent results reported among the studies may be ascribed to the difference in supplement concentration, additive form, and additive self-characteristics.

During storage, comparatively higher HU value was observed in the eggs of FA-fed hens stored for 3 weeks. Lee et al. (2017) reported increased total antioxidant activity with decreased content of polyphenolic and flavonoid in L. plantarum-fermented A. annua L. Some substances newly-synthesized during fermentation may be more effective to prevent egg albumen loss during storage at room temperature (Ibrahim et al., 2014; Dong et al., 2015). Fermented Ginkgo-leaves have been reported to increase monounsaturated fatty acids, polyunsaturated fatty acids (PUFA), and the ratio of PUFA/saturated fatty acids in the egg yolk (Zhao et al., 2013). The increased content of unsaturated fatty acids is considered to be more efficient to improve egg oxidative stability and extend egg freshness period. In the present study, both NFA and FA additives were effective in reducing MDA concentration in egg yolk, which is consistent with previous studies that verified MDA reduction in the meat of broilers fed different Artemisia species (Kim et al., 2012a; Kim et al., 2012b; Cherian et al., 2013). In addition, the dietary supplementation of plant extract mixture from A. capillaris, Camellia sinensis, Schizandrachinensis, and Viscum album var. coloratum has increased HU of egg stored for 2 and 3 weeks, coupled with a decrease in yolk MDA concentration. In addition of its most well-known compound, artemisinin, 80 natural products have been identified in Artemisia annua, out of which phenolics are the most abundant active compounds, which have diverse antioxidant, antimicrobial, and anti-inflammatory effects (Czechowski et al., 2018, Lee et al., 2017; Kim et al., 2015). Overall, the effects of the supplemented additives did not influence laying performance, except for egg weight. In comparison with L. plantarum and non-fermented A. annua, fermented A. annua showed better effect on changing eggshell color. In addition, both FA and NFA supplementation showed oxidative protective effect on egg yolk during storage.

CONCLUSION

This is the first study determining the effects of fermented and non-fermented A. annua dried leaves as phytogenic feed additives on laying performance, egg quality, serum cholesterol, and lipid oxidation in 40-week-old Hy-Line Brown layers. A synergistic beneficial effect was observed by the supplementation of fermented A. annua L. with L. plantarum (FA) compared to non-fermented A. annua L. (NFA) and L. plantarum only (LO) in the basal diet. In the future work, it is worthy to investigate the dietary effect of non-fermented and fermented A. annua L. on egg lipid composition as well as on the immunity of laying hens.

ACKNOWLEDGMENT

This work was supported by Korean Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through Agri-Bioindustry Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (Project No. 118051-03).

REFERENCES

• Abad MJ, Bedoya LM, Apaza L, Bermejo P. The Artemisia L. genus: a review of bioactive essential oils. Molecules 2012;17(3):2542-2566.
• Abd El-Hack ME, Mahgoub SA, Alagawany M, Ashour EA. Improving productive performance and mitigating harmful emissions from laying hen excreta via feeding on graded levels of corn DDGS with or without Bacillus subtilis probiotic. Journal of Animal Physiology and Animal Nutrition 2017;101(5):904-913.
• Abdel-Wareth AAA, Lohakare JD. Effect of dietary supplementation of peppermint on performance, egg quality, and serum metabolic profile of Hy-Line Brown hens during the late laying period. Animal Feed Science and Technology 2014;197:114-120.
• Abou-Elkhair R, Selim S, Hussein E. Effect of supplementing layer hen diet with phytogenic feed additives on laying performance, egg quality, egg lipid peroxidation and blood biochemical constituents. Animal Nutrition 2018;4(4):399-400.
• Almeida GF, Horsted K, Thamsborg SM, Kyvsgaard NC, Ferreira JF, Hermansen JE. Use of Artemisia annua as a natural coccidiostat in free-range broilers and its effects on infection dynamics and performance. Veterinary Parasitology 2012;186(3-4):178-187.
• Aydin R, Karaman M, Cicek T, Yardibi H. Black cumin (Nigella sativa L.) supplementation into the diet of the laying hen positively influences egg yield parameters, shell quality and decreases egg cholesterol. Poultry Science 2008;87:2590e5.
• Baghban-Kanani P, Hosseintabar-Ghasemabad B, Azimi-Youvalari S, Seidavi A, Ragni M, Laudadio V, et al. Effects of using artemisia annua leaves, probiotic blend, and organic acids on performance, egg quality, blood biochemistry, and antioxidant status of laying hens. The Journal of Poultry Science 2018;56(2):120-127.
• Bora KS, Sharma A. The genus Artemisia: a comprehensive review. Pharmaceutical Biology 2011;49(1):101-109.
• Bosselmann AS, Gylling M. Economic potential of a Danish production of Artemisia annua based feed additives for broilers [IFRO report, 224]. Frieleriksbey: Department of Food and Resource Economics, University of Copenhegan; 2013.
• Botsoglou NA, Fletouris DJ, Papageorgiou GE, Vassilopoulos, Mantis AJ, Trakatellis AG. Rapid, sensitive, and specific thiobarbituric acid method for measuring lipid peroxidation in animal tissue, food, and feedstuff samples. Journal of Agricultural and Food Chemistry 1994;42(9):1931-1937.
• Brisibe EA, Umoren UE, Owai PU, Brisibe F. Dietary inclusion of dried Artemisia annua leaves for management of coccidiosis and growth enhancement in chickens. African Journal of Biotechnology 2008;7(22):4083-4092.
• Brown GD. The biosynthesis of artemisinin (Qing haosu) and the phyto-chemistry of Artemisia annua L. (Qinghao). Molecules 2010;15(11):7603-7698.
• Butler MW, McGraw KJ. Eggshell coloration reflects both yolk characteristics and dietary carotenoid history of female mallards. Functional Ecology 2013;27:1176-1185.
• Cherian G, Orr A, Burke IC, Pan W. Feeding Artemisia annua alters digesta pH and muscle lipid oxidation products in broiler chickens. Poultry Science 2013;92(4):1085-1090.
• Choi E, Park H, Lee J, Kim G. Anticancer, antiobesity, and anti-inflammatory activity of Artemisia species in vitro. Journal of Traditional Chinese Medicine 2013;33(1):92-97.
• Czechowski T, Larson TR, Catania TM, Harvey D, Wei C, Essome M, Brown GD, Graham IA. Detailed phytochemical analysis of high-and low artemisinin-producing chemotypes of Artemisia annua. Frontiers in Plant Science 2018;9:641.
• Di Cagno R, Coda R, De Angelis M, Gobbetti M. Exploitation of vegetables and fruits through lactic acid fermentation. Food Microbiology 2013;33(1):1-10.
• Dong JW, Cai L, Xiong J, Chen XH, Wang WY, Shen N, et al. Improving the antioxidant and antibacterial activities of fermented Bletillastriata with Fusarium avenaceum and Fusarium oxysporum. Process Biochemistry 2015;50(1):8-13.
• Engberg RM, Grevsen K, Ivarsen E, Fretté X, Christensen LP, Højberg O, et al. The effect of Artemisia annua on broiler performance, on intestinal microbiota and on the course of a Clostridium perfringens infection applying a necrotic enteritis disease model. Avian Pathology 2012;41(4):369-376.
• Forte C, Moscati L, Acuti G, Mugnai C, Franciosini MP, Costarelli S, et al. Effects of dietary Lactobacillus acidophilus and Bacillus subtilis on laying performance, egg quality, blood biochemistry and immune response of organic laying hens. Journal of Animal Physiology and Animal Nutrition 2016;100(5):977-987.
• Fraser GE. Vegetarian diets: what do we know of their effects on common chronic diseases? The American Journal of Clinical Nutrition 2009;89(5):1607S-12S.
• Haugh RR.The Haugh unit for measuring egg quality. United States Egg and Poultry Magazine 1937;43:552-555.
• Hossain ME, Ko SY, Kim GM, Firman JD, Yang CJ. Evaluation of probiotic strains for development of fermented Alisma canaliculatum and their effects on broiler chickens. Poultry Science 2012;91(12):3121-3131.
• Ibrahim NA, Mustafa S, Ismail A. Effect of lactic fermentation on the antioxidant capacity of Malaysian herbal teas. International Food Research Journal 2014;21(4):1483-1488.
• Khalaji S, Zaghari M, Hatami KH, Hedari-Dastjerdi S, Lotfi L, Nazarian H. Black cumin seeds, Artemisia leaves (Artemisia sieberi), and Camellia L. plant extract as phytogenic products in broiler diets and their effects on performance, blood constituents, immunity, and cecal microbial population. Poultry Science 2011;90(11):2500-2510.
• Khan SH, Anjum MA, Parveen A, Khawaja T, Ashraf NM. Effects of black cumin seed (Nigella sativa L.) on performance and immune system in newly evolved crossbred laying hens. Veterinary Quarterly 2013;33:13e9.
• Kim CH, Kim GB, Chang MB, Bae GS, Paik IK, Kil DY. Effect of dietary supplementation of Lactobacillus-fermented Artemisia princeps on growth performance, meat lipid peroxidation, and intestinal microflora in Hy-line Brown male chickens. Poultry Science 2012a;91(11):2845-2851.
• Kim DW, Kim JH, Kang GH, Kang HK, Choi JY, Kim SH, et al. Effects of water extract mixtures from Artemisia capillaris, Camellia sinensis, Schizandra chinensis, and Viscum album var. Coloratum on laying performance, egg quality, blood characteristics, and egg storage stability in laying hens. Korean Journal for Food Science of Animal Resources 2010;30(3):449-457.
• Kim WS, Choi WJ, Lee S, Kim WJ, Lee DC, Sohn UD, et al. Anti-inflammatory, antioxidant and antimicrobial effects of artemisinin extracts from Artemisia annua L. The Korean Journal of Physiology and Pharmacology 2015;19(1):21-27.
• Kim YJ, Kim CM, Choi JH Choi IH. Effect of dietary mugwort (Artemisia vulgaris L.) and pine needle powder (Pinusdensiflora) on growth performance, serum cholesterol levels, and meat quality in broilers. African Journal of Biotechnology 2012b;11(55):11866-11873.
• Kooy F, Sullivan SE. The complexity of medicinal plants: The traditional Artemisia annua formulation, current status and future perspectives. Journal of Ethnopharmacology 2013;150(1):1-13.
• Lee AR, Niu KM, Kang SK, Han SG, Lee BJ, Kim SK. Antioxidant and antibacterial activities of Lactobacillus-fermented Artemisia annua L. as a potential fish feed additive. Journal of Life Science 2017;27(6):652-660.
• Lei XJ, Yun HM, Kim IH. Effects of dietary supplementation of natural and fermented herbs on growth performance, nutrient digestibility, blood parameters, meat quality and fatty acid composition in growing-finishing pigs. Italian Journal of Animal Science 2018;17(4):984-993.
• Loh TC, Choe DW, Foo HL, Sazili AQ, Bejo MH. Effects of feeding different postbiotic metabolite combinations produced by Lactobacillus plantarum strains on egg quality and production performance, faecal parameters and plasma cholesterol in laying hens. BMC Veterinary Research 2014;10(1):149.
• Lokaewmanee K, Yamauchi K, Thongwittaya N. Effects of fermented plant product on growth performance, some blood variables, carcase characteristics, and intestinal histology in broilers. British Poultry Science 2012;53(2):215-223.
• Mazanko MS, Gorlov IF, Prazdnova EV, Makarenko MS, Usatov AV, Bren AB, et al. Bacillus probiotic supplementations improve laying performance, egg quality, hatching of laying hens, and sperm quality of roosters. Probiotics and Antimicrobial Proteins 2018;10(2):367-373.
• Mohammadi Gheisar M, Kim IH. Phytobiotics in poultry and swine nutrition-a review. Italian Journal of Animal Science 2018;17(1):92-99.
• Monira KN, Salahuddin M, Miah G. Effect of breed and holding period on egg quality characteristics of chicken. International Journal of Poultry Science 2003;2(4):261-263.
• Ng CC, Wang CY, Wang YP, Tzeng WS, Shyu YT. Lactic acid bacterial fermentation on the production of functional antioxidant herbal Anoectochilus formosanus Hayata. Journal of Bioscience and Bioengineering 2011;111:289-293.
• Nishizawa N, Fudamoto Y. The elevation of plasma concentration of high-density lipoprotein cholesterol in mice fed with protein from proso millet. Bioscience, Biotechnology, and Biochemistry 1995;59(2):333-335.
• Niu KM, Kothari D, Cho SB, Han SG, Song IG, Kim SC, et al. Exploring the probiotic and compound feed fermentative applications of Lactobacillus plantarum SK1305 isolated from Korean green chili pickled pepper. Probiotics and Antimicrobial Proteins 2018;1-12.
• Peres CM, Peres C, Hernández-Mendoza A, Malcata FX. Review on fermented plant materials as carriers and sources of potentially probiotic lactic acid bacteria-with an emphasis on table olives. Trends in Food Science & Technology 2012;26(1):31-42.
• Qin S, Hou DX. The biofunctions of phytochemicals and their applications in farm animals: the nrf2/keap1 system as a target. Engineering 2017;3(5):738-752.
• Samiullah S, Roberts JR, Chousalkar K. Eggshell color in brown-egg laying hens - a review. Poultry Science 2015;94(10):2566-2575.
• Song Z, Cheng K, Zhang L, Wang T. Dietary supplementation of enzymatically treated Artemisia annua could alleviate the intestinal inflammatory response in heat-stressed broilers. Journal of Thermal Biology 2017;69:184-190.
• Tsai YT, Cheng PC, Pan TM. The immunomodulatory effects of lactic acid bacteria for improving immune functions and benefits. Applied Microbiology and Biotechnology 2012;96(4):853-862.
• Wan XL, Jiang LY, Zhong HR, Lu YF, Zhang LL, Wang T. Effects of enzymatically treated Artemisia annua L. on growth performance and some blood parameters of broilers exposed to heat stress. Animal Science Journal 2017;88(8):1239-1246.
• Wan XL, Niu Y, Zheng XC, Huang Q, Su WP, Zhang JF, et al. Antioxidant capacities of Artemisia annua L. leaves and enzymatically treated Artemisia annua L. in vitro and in broilers. Animal Feed Science and Technology 2016;221a:27-34.
• Yalçin S, Yalçin S, Erol H, Bugdayci KE, Özsoy B, Çakir S. Effects of dietary black cumin seed (Nigella sativa L.) on performance, egg traits, egg cholesterol content and egg yolk fatty acid composition in laying hens. Journal of the Science of Food and Agriculture 2009;89(10):1737-1742.
• Zhang JL, Xie QM, Ji J, Yang WH, Wu YB, Li C, Ma JY, et al. Different combinations of probiotics improve the production performance, egg quality, and immune response of layer hens. Poultry Science 2012;91(11):2755-2760.
• Zhao L, Zhang X, Cao F, Sun D, Wang T, Wang G. Effect of dietary supplementation with fermented Ginkgo-leaves on performance, egg quality, lipid metabolism and egg-yolk fatty acids composition in laying hens. Livestock Science 2013;155(1):77-85.

Publication Dates

• Publication in this collection
  20 Dec 2019
• Date of issue
  2019

History

• Received
  23 Nov 2018
• Accepted
  27 June 2019