Effects of mangosteen pericarp extraction on egg production, stress index and yolk cholesterol in laying quails

W Boontiam and S Kittipongpysan¹

Faculty of Agriculture, Department of Animal Science, Khon Kaen University, Khon Kaen, Thailand 40002
¹ Faculty of Agriculture and Life Science, Department of Agricultural Science and Entrepreneurship, Chandrakasem Rajabhat University, Bangkok, Thailand 10900
sumetee.k@chandra.ac.th

Abstract

This study investigated the effects of crude mangosteen pericarp ( Garcinia mangostana) extract with different dilutions on egg production, egg quality, stress response, and yolk cholesterol in laying quails. Two hundred and forty laying quails were divided into four diluting treatments. Each group had three replications with 20 quails in each. The treatments included a control group without dilution, and the remaining treatments were diluted with water (mangosteen pericarp : water) at the ratios of 1:30, 1:20, and 1:10, respectively. Results showed that the dilution of mangosteen pericarp extract minimized damaged eggs (p = 0.036), whereas improved albumen height (p = 0.003), Haugh unit (p = 0.025), and eggshell thickness (p = 0.073). Furthermore, the heterophil-to-lymphocyte ratio (p = 0.071) and yolk cholesterol (p = 0.025) decreased in response to the mangosteen pericarp extract dilutions. No significant differences for water intake, egg production, egg weight, egg mass, feed dozen eggs per kg of egg produced, feed consumption, livability percentage, and yolk color score. To conclude, the dilution of MCE in drinking water may be an alternative approach in promoting egg and eggshell quality and lowering yolk cholesterol for consumer preference.

Key words: drinking water, heat stress, lipid metabolism, mangosteen peel

Introduction

Heat stress (HS) is an important consideration for the poultry industry. Previous studies showed that the increased temperature above 36 ^oC for 10 hours daily caused the activation of hypothalamic-pituitary-adenal axis and corticosterone secretions in poultry (Star et al 2008, Quinteiro-Filho et al 2010). Corticosterone is a well-established stress hormone in poultry and functions in immunosuppression and impaired animal performance (Honda et al 2015). Rozenboim et al (2007) found that the increased temperature at 42 ^oC impaired ovarian function by inhibiting the secretion of steroidogenic enzyme. A previous report also found that the regulation of acid-base decreased as birds were exposed to heat stressors because of poor eggshell quality (Lin et al 2004). To solve this problem, the mangosteen ( Garcinia mangostana) pericarp extraction (MCE) was included in the tap water.

Mangosteen (Garcinia mangostana) is commonly found in Indonesia, Philippines, Myanmar, and Thailand (Pedraza-Chaverri et al 2008) and contains various active substances of phenolic compounds, especially anthocyanin and xanthones (Chen et al 2008; Zadernowski et al 2009). Mangosteen prevents bacterial invasion (Suksamran et al 2003) and is antitumor (Yu et al 2009), anti-inflammatory (Chen et al 2008), and antioxidant (Mohamed et al 2014; Thong et al 2015). The in-feed supplementation of mangosteen pericarp powder at 1 g per kg diet has positive effects on eggshell quality, while lowering yolk cholesterol in laying birds (Rusli et al 2015). However, there is little research on the application of crude mangosteen pericarp extract in the drinking water for laying quails. Consequently, we aimed to examine the effects of various rations of MCE in drinking water on egg production and quality, heterophil-to-lymphocyte ratios (H:L ratios), and yolk cholesterol of laying quails.

Materials and methods

Laying quails, housing, and experimental diet

A total of 240 laying quails were allotted into 4 treatments based on the average egg rate (82.38%). Each group consisted of three replicates with 20 quails, performing under a completely randomized randomize design. The treatments were provided different MCE dilutions with tap water in the following rations: control treatment without the addition of MCE (MCE0), and the remaining treatments included MCE in the tap water at concentrations of: ratios of 3% (MCE3), 5% (MCE5) and 10%(MCE10). The experiment lasted 9 weeks. The MCE was extracted as follows:  (Tjahjani et al 2014).

The experiment was conducted in the central region of Thailand (Ang Thong, Thailand). The quails were raised in-controlled temperature housing in a wire-floor cage (width 20 cm x length 22 cm x height 17 cm) with individual nippers. Artificial light was provided from 0400 h). The birds freely accessed feed and water during the experimental period.

The mash form of experimental diets was formulated to meet or exceed the recommendation of NRC (1994) for laying quails (Table 1). We followed the instruction of the National Research Council (Bangkok, Thailand) for animal care and handling.

Egg production, feed conversion and water consumption

Eggs were collected and weighed daily for the determinations of hen-day egg production and average egg weight. Egg mass was calculated according to this equation; (hen-day egg production x average egg weight)/100. Water intake and wasted water were recorded twice daily, and expressed as the average value of 9 weeks. The data were used to calculate feed dozen per kilogram of egg produced, and feed consumption. Number of laying quails dead in each replicate were used to adjust feed consumption, and indicate livability percentage. The percentage of damaged eggs (hairline crack, pinhole crack and body crack eggs) was recorded.

Egg quality analyses

We randomly collected eggs, using 30 eggs per treatment (n = 90 samples) in the 3-week interval for the determination of yolk color score, albumen height, and eggshell thickness. We broke individuals to measure the yolk color score using a Roche color fan and albumen height using an albumen height gauge (Technical Services and Supplies, York, UK). Further, we used the value of albumen height to determine the Haugh unit, using the equation of Eisen et al (1962). We determined the eggshell thickness after removing the adhered albumen, washing the eggshell with tap water, and drying for 48 h at room temperature. We measured the eggshell thickness three times at the egg’s equator using a digital micrometer (Mitutoyo Corporation, Kanagawa, Japan) and reported the average value.

Blood collection and analyses

Blood collection was from randomly selected three birds per replication at the end of the experiment from 0800 to 0900 of approximately 1.5 mL each using sterilized needles and syringes via a jugular vein. We immediately transferred the samples into 3-mL anticoagulant tubes and determined heterophil-to-lymphocyte ratios (H/L ratios) using the Gross and Siegel (1983) method. We immediately counted 100 heterophils and the presence of lymphocytes on each slide for each quail using a light microscope (Olympus American, Center Valley, PA).

Yolk collection and yolk cholesterol analyses

At the end of experiment, we randomly chose 48 eggs (using four eggs for each replication) for a yolk cholesterol assay. We separated the egg yolk from the egg white and prepared a further assay using a method established by Pasin et al (1998). We analyzed the egg yolk cholesterol using commercial kits. We presented the cholesterol concentration in the egg yolk as a unit of mg per g of egg yolk.

Statistical analysis

Data were analyzed for laying performance, egg quality, H:L ratio, and yolk cholesterol according to a completely randomized design using the general linear model (GLM) procedure of the SAS software (SAS Inst., Inc., Cary, NC, USA). Linear and quadratic effects of MCE dilutions were performed using orthogonal polynomial contrasts. A significant value was detected at p < 0.05 or p< 0.10.

Results

Laying performance

Treatments had no influence on water intake, egg production, egg weight, egg mass, feed per kilogram of egg produced, feed consumption, and livability. The percentage of damaged eggs decreased linearly as the MCE concentration increased with a cuadratic response (Figure 1),

Egg quality

In all periods, the MCE concentration did not affect the yolk color score but increased albumen height, HU, and eggshell thickness. The MCE improved HU and eggshell thickness over the whole period.

Stress index and yolk cholesterol

The H/L ratio and cholesterol content in egg yolk showed a linear decrease in response to  MCE concentration (Table 4). The cholesterol was reduced from 16.6 to 13.0 mg per g of egg yolk.

Discussion

The increasing dilution of MCE improved eggshell quality by decreasing the damaged eggs and increasing the eggshell thickness. The positive effect may be caused by the presence of phenolic compounds in MCE, which reacts with oxygen before attacking macromolecules (Maqsood et al 2015; Mohamed et al 2014; Tjahani et al 2014). This might maintain homeostasis without impairing the egg production and quality. However, we observed poor eggshell quality in the control treatment without MCE. A previous study showed that poultry are more likely to be susceptible to respiratory alkalosis (Borges et al 2007). It subsequently lowered the carbon dioxide in the blood, which is a major substance for bicarbonate synthesis (Borges et al 2007). The imbalance of this substance negatively influenced eggshell quality. The current study also found that the dilution of MCE increased the quality of albumen freshness, as indicated in albumen height and HU measurements. A possible explanation is the higher amount of phenolic compounds that inhibiting protein degradation in the egg white (Maqsood et al 2015). However, in the current study, we did not observe significant increases in yolk color scores in the MCE-supplemented treatments. This may be the result of the mangosteen pericarp extraction method. From these results, the MCE dilution could be an alternative approach for improved egg and eggshell quality

The heterophil-to-lymphocyte ratio (H/L) is a well-established indication of significant body stress in poultry. Birds typically maintain homeostasis through the increased secretion of glucocorticorticoids (Gross and Siegel 1983). This mechanism suppressed the functions of macrophage (Quinteiro-Filho et al 2010) and subsequently changed the H:L ratio during chronic stress (López-Olvera et al 2007). The increased heterophils caused lower production of lymphocytes and greater secretion of inflammatory cytokines (Harmon et al 1998). The reduction of H:L in the birds consuming MCE might suppress the migration of heterophil granuloctes into the blood circulation. Therefore, the lower H:L ratio, as a lower heat stress, could alleviate the stress of laying quails.

Previous reports demonstrated that the dietary supplementation of mangosteen pericarp in a 1 g per kg diet significantly reduced yolk cholesterol concentrations (Rusli et al 2015). This is consistent with our finding that the yolk cholesterol linearly decreased from 16.55 to 12.99 mg per g of yolk when offered MCE in the drinking water. Chen et al (2008) and Zadernowski et al (2009) found that the mangosteen is enriched by xanthones, α-mangostin, and tannin. Studies reported active compounds in regulating cholesterol biosynthesis via the suppression of hydroxy-methyl glutaryl CoA reductase (HMG-CoA reductase). The lower production of HMG-CoA reductase will activate the catabolism of low-density lipoproteins and cholesterol concentrations before use in cholesterol synthesis. Therefore, providing MCE in the drinking water could be an alternative approach in producing low egg yolk cholesterol in representing a health benefit for consumers.

Conclusion

• The dilution of MCE significantly increased  eggshell quality and decreased yolk cholesterol, without impairing egg production and feed efficiency of the laying quails.
• The MCE dilution could alleviate stress in the laying quail via lowered heterophil-to-lymphocyte ratios.
• This approach could be the solution for quail production in the tropical country.

Acknowledgement

The authors gratefully thank Chandrakasem Rajaphat University (Bangkok, Thailand) for the financial support, and sincerely thank Anek Farm (Ang Thong, Thailand) for animal facilities.

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