Evaluation of Pelleting Mealworm (Tenebrio Molitor) Powder with Chokeberry By-product Meal To improve Duck Production and Meat Quality Traits

ABSTRACT This study aimed to evaluate the effects of dietary supplementation pelleted Tenebrio molitor (TM) powder with chokeberry by-product (CBP) meal on duck production and meat quality traits. A total of 180 0-day-old Pekin ducks were allocated randomly between three dietary groups viz., control, Treatment 1, and Treatment 2, with three pens per group and twenty birds per pen, characterized by increasing levels of pelleted TM powder with CBP meal in concentration of 0%, 1.5%, and 3%, respectively, and reared until 42 days of age in a completely randomized design. On day 42, the ducks were sacrificed, and breast and thigh muscles were used for meat quality evaluations. Feeding pelleted TM powder with CBP meal resulted in a significant difference in final body weight, weight gain and feed conversion ratio (p<0.05). Additionally, a significant difference was observed in the pH of the breast meat samples (p<0.05). As the only significant difference between the breast and thigh meat, 1,1-diphenyl-2-picryhydrazyl radical scavenging was influenced significantly by the dietary inclusion of pelleted TM powder with CBP meal (p<0.05). However, the values of thiobarbituric acid reactive substances and meat color in both breast and thigh meat were insignificant (p>0.05) different among all groups. To conclude, the inclusion of up to 3 % of pelleted TM powder with CBP meal in the diets of ducks did improve the growth production and antioxidant characteristics effects, which could decrease TBARS and reflected by changes in 1,1-diphenyl-2- picrylhydrazyl free DPPH radical scavenging values.


INTRODUCTION
Increasing demand for animal products is related to rapid human population growth and urbanization.Animal feeding and nutrition play an essential role in animal production, including economic and social roles, thus providing multiple functions (Yosef et al., 2022).They are part of an animal product-system chain between crop cultivation and the production and processing of protein-rich animal products (Yosef et al., 2022).In particular, as feed accounts for 60-70% of the total cost in poultry production, any operation to optimize feed efficiency or reduce the feed cost may lead to a reduction in feed ingredient prices or the total cost of production (Thirumalaisamy et al., 2016).Pelleting provides for one such way of optimizing feed efficiency while reducing the feed cost.In addition, it also offers an effective way of mixing insect powders and meals of plant origin.Using insects has been considered an alternative source of conventional protein feed to soybean meal and fish meal or decomposition abilities from animal wastes (Choi, 2022).Especially, other studies have shown the positive effects of using insects as animal diets on poultry production and meat quality or antioxidant (Ojewola et al., 2002;Choi et al., 2021a, b, c; Choi IH

Evaluation of Pelleting Mealworm (Tenebrio Molitor) Powder with Chokeberry By-product Meal To improve
Duck Production and Meat Quality Traits Jeong et al., 2022).Tenebrio molitor (TM), well-known mealworm, represents one of the most interesting edible insects studied as feed and food as it can be easily reared and maintained at early stages and also due to its larval size (Ghaly & Alkoaik, 2009;Morales-Ramos et al., 2012).Chokeberry is cultivated mainly in the east-south of Europe as an industrial crop and has been traditionally used in North America as medicine (Seidemann, 1993;Kokotkiewicz et al., 2010).Due to its antioxidant and beneficial effects on vitamin action, chokeberry has become even more popular in recent years (Denev et al., 2012).In addition, chokeberry is used as an ingredient in juices, wines, and jams and as a coloring agent and nutritional supplement (Kulling & Rawel, 2008).However, the disposal of chokeberry waste is a major environmental problem.Pelleting of chokeberry by-product (CBP) meal reduces food waste and helps in recycling in poultry industries thereby offering a solution to the problem of chokeberry waste disposals.In this context, the use of pelleted TM powder with CBP meal offers a method for improving poultry production and meat quality by decreasing feed wastage (Jeong et al., 2022).In the present study, the effects of dietary supplementation of pelleted TM powder and CBP meal on growth characteristics and meat quality in Pekin ducks was investigated.Thus, the results presented in this study provide pelleting as an alternative technique for mixing insect powder and CBP meal in duck diets.

Sample preparation
Tenebrio molitor (TM) powder was obtained from Modnilove (Ulju, South Korea).Chokeberry by product was provided by Yusim Farming Asscociation (Yeongju, Korea).To prepare a CBP meal, the wet CBP was airdried for seven days and kept away from urban traffic to ensure appropriate protection against environmental contamination, followed by hot-air drying at 60°C for 24 h (Kim et al., 2010).After drying, the CBP was ground, sieved, and drilled with 1-mm trapezoid holes.TM powder and chokeberry meal were thoroughly mixed in the ration of 7:3, and pellets were prepared to form a mixture of TM powder and chokeberry meal using a pelleting mill (Kum Kang Eng., Daegu, South Korea).

Animal Husbandry
The protocol for the animal trial complied with the animal care guidelines of animal policy by the Gilhong farm (Geochang, South Korea).A total of 180 0-day-old Pekin ducks were allocated randomly into three groups (n = 45) viz., Control, Treatment 1 and Treatment 2 fed pelleted TM powder with CBP meal in concentration of 0%, 1.5%, and 3%, respectively, with three replicate pens of twenty birds each in a completely randomized design.The basal diets were used during the starter (0-21 d, 21.0%CP, 0.4% Ca, and 1.5% P) and grower phases (22-42 d, 17.0% CP, 0.4% Ca, and 1.0% P).All ducks were maintained under an age-appropriate temperature, light schedule, ventilation and humidity, with water and diet available ad libitum.To determine final body weight (FBW), body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR), ducks were weighed at 0 d and 42 d of age, with feed weighed regularly every seven days.

Slaughter processing
At 42 d of age, 36 ducks with four ducks per pen were selected for slaughter processing, following 12 h fasting.The selected ducks were transported to the slaughterhouse to be electrically stunned and slaughtered by a ventral cut of neck blood vessels.After bleeding, carcasses were scalded at 60 ºC for 2 min, plucked, and eviscerated manually.Duck breast and thigh samples were cleared of all skin, subcutaneous fat, and visible connective tissue, placed in a resealable plastic bag, and immediately stored at 4 °C.

Meat quality traits
Meat samples were measured for pH, thiobarbituric acid reactive substances (TBARS), 1,1-diphenyl-2picrylhydrazyl (DPPH) radical scavenging activity, and meat color.Muscle pH was analyzed for homogenates of 10 g sample in 90 mL of distilled water using a pH-meter (Mettlerr Toledo Co, MP 230, Greifensee, Switzerland).A TBARS assay was performed according to the method described by Sinnhuber and Yu (1977).The absorbance was measured at 532 nm by spectrophotometer (UV-mini-1240, Shimadzu, Japan).The results were calculated as milligrams of malonaldehyde per kilogram of meat.The DPPH radical scavenging activity of meat samples was determined according to the method described by Blois (1958).Absorbance was measured with a UV-visible spectrophoto meter (UV-mini-1240, Shi madzu Corp., Kyoto, Japan) set at 515 nm.Scavenging activity was calculated based on the change in absorbance.The ascorbic acid and α-tocopherol (Sigma, St. Louis, MO, USA) were used as the positive control.Meat color (CIE L*a* b*) was measured from five random locations of the sample surface using a Minolta Chromameter (CR-

Statistical Analysis
The data were analyzed by standard analysis of variance (ANOVA) using SAS GLM procedures (SAS Institute Inc., 2002) with the pen used as the experimental unit.Duncan's multiple range procedures considering p<0.05 as significant were used for comparisons among means.

RESULTS
Table 1 showed the effects of pelleted TM powder with CBP meal on the growth production of ducks after six weeks.Dietary supplementation of pellets of TM powder with CBP meal resulted in significant differences in FBW, BWG and FCR (p<0.05), in the absence of differences in the initial body weight and feed intake between the control and treatment groups in Pekin ducks.Compared with the control groups, the T1 and T2 treatments displayed better Mean in the same column with no common superscript are significantly different (p<0.05).
2 Values are expressed as mean ± standard error.
2 Values are expressed as mean ± standard error.
duck production measured in terms of FBW, BWG and FCR.In addition, a diet containing 1.5% and 3% pelleted TM powder with CBP meal (T1 and T2) to ducks resulted in a similar tendency for growth performance.
Effects of pelleted TM powder with CBP meal on meat quality traits of ducks at 42 days are presented in Table 2.A significant difference was observed in pH values in the breast meat samples (p<0.05).As for the meat quality traits of the thigh meat, pH was not significantly (p>0.05)different among all groups.TBARS values in the breast and thigh meat were unaffected by the inclusion of pelleted TM powder with CBP meal (p>0.05).As the only difference between the breast and thigh meat, DPPH radical scavenging was influenced significantly by the dietary inclusion of pelleted TM powder with CBP meal (p<0.05).The meat color in the breast and thigh meat were not affected by dietary supplementation of pelleted TM powder with CBP meal (p>0.05). eRBCA-2022-1717

DISCUSSION
Overall, our results were consistent with the results reported by Elahi et al. (2020), herein feeding 4% mealworm meals to Ross 308 male broiler chickens resulted in increased body weight, average daily gain, and FCR in the starter phase.Hussain et al. (2017) reported that chickens fed a diet containing 3% mealworm meals, showed increase in weight gain and dressing percentage, including feed cost and economic profit.In terms of using chokeberry, Lee et al. (2018) used the level of 1% chokeberry powder in the diet of ducks and found improvement in the weight gain and the feed:gain ratio.However, no study has so far evaluated the impact of a pelleting combination of TM powder and CBP meal on the growth performance of Pekin ducks.Based on these results, a wide range of alternative feedstuffs is available for these two feed materials (Tenebrio molitor powder and chokeberry byproduct meal) in duck production systems.In addition, the duck feed industry requires a constant supply of any newer feed resource with consistent quality while making the best use of constantly changing raw materials (Thirumalaisamy et al., 2016).In other words, there is an opportunity for on-farm mixing feeds with locally alternative feedstuffs.In the present study, the possible mechanisms for improving the growth performance of ducks could be a result of two reasons: 1) pelleting effects (high digestibility) that the ducks receive a balanced supply of nutrients (Abdollahi et al., 2019), and 2) the interaction of bioactive compounds (for example, phenolic constituents) present in TM powder and CBP meal (Kulling & Rawel, 2008).Consequently, it is hypothesized that there is possibly evidence from the current study for a pelleting and bioactive effect of a mixture of TM powder and CBP meal in ducks.
For meat quality traits, the pH values of the breast and thigh meat obtained in the present trial showed different patterns.The pH values of the breast meat were decreased by increasing pelleted TM powder with CBP meal compared with control.In general, the decline in meat pH is due to the change in glycogen decomposition rate caused by the formation of lactate (Bendall, 1960), which could be ascribed to the rapid growth and hypertrophy of muscles in ducks by an interaction effect of pelleted TM powder and CBP meal, especially in the breast as described by Popova et al. (2020) andFernández-López et al. (2005).It has also been reported that broiler quails fed diets containing 15% black soldier fly meal result in a decrease in meat pH, suggesting that the addition of TM powder and CBP meal to diets can directly influence the pH of meat (Cullere et al., 2016;Kim et al., 2016).On the other hand, the pH values in thigh meat were similar among groups.However, the majority of the studies in poultry did not observe the change in the meat pH using TM.It might have resulted from using moderate to high levels of yellow meal worm in the diet (Cullere et al., 2016;Secci et al., 2018).The TBARS results of the breast meat were the lowest in T1, followed by T2 and control.In addition, the reduction in the TBARS results of the thigh meat was in the following order: T2 > T1 > Control.The data obtained from the current study suggested that the reason for reducing the TBARS content in both breast and thigh meat was chiefly related to a pelleting effect or the effective antioxidants that are transferred from feed to animal tissue by TM powder and CBP meal (Kulling & Rawel, 2008;Jeong, 2022).The DPPH radical scavenging in both breast and thigh meat of ducks as antioxidant effectiveness decreased in the order: Control > T1 > T2.Our observation is consistent with previous studies on broiler chickens using Hermetia illucens and Protaetia brevitarsis seulensis powder as dietary supplements (Choi et al., 2021c).According to Jeong (2022), adding of 1.5% and 3% Ptecticus tenebrifer to duck diets improved DPPH radical scavenging values compared with the controls.A further possible reason for DDPH effects has been that the supplementation of pelleted TM powder with CBP meal results in the presence of biologically active substances (antioxidants effect) that also may interact.The overall color traits obtained in our study are similar to those reported by Schiavone et al. (2019), wherein dietary supplementation of Hermetia illucens meal did not affect the meat color in the breast or thigh meat of broiler.In general, the color of food and meat products are a crucial attribute considered by consumers and used as an economic index of foodstuff (Qiao et al., 2001).However, the cause for the lack of change in meat color in response to the dietary supplementation of pellets of TM powder with chokeberry meal remains unexplained.
In conclusion, the results of the present study provide meaningful and innovative information on the effects of supplementation of partially pelleted TM powder with CBP meal in duck diets.The dietary inclusion of 1.5% and 3% pelleted TM powder with CBP meal increased final body weight, body weight gain, and feed:gain ratio.The comparable results, in terms of meat quality traits and antioxidant effects, between the pelleted TM powder with CBP meal and the control group demonstrated a decrease in TBARS and an increase in DPPH radical scavenging in the eRBCA-2022-1717

Evaluation of Pelleting Mealworm (Tenebrio Molitor) Powder with Chokeberry By-product Meal To improve Duck Production and Meat Quality Traits
breast and thigh meat of ducks.Future research is still needed to investigate the effect of pelleted TM powder with CBP meal on duck breast and thigh meat colors, including various meat quality traits.

Table 1 -
Effects of pelleted TM powder with CBP meal on duck production after 42 days.

Table 2 -
Effects of pelleted TM powder with CBP meal on meat quality traits of ducks at 42 days.