| Artemisia annua L.(A.annua)is a natural herb belongs to Artemisia species of Asteraceae and has been used in folk medicine for therapeutic purpose including treating fever and malaria.In addition to artemisinin,A.annua has essential oils,phenolics,flavonoids,as well as amino acids,vitamins and mineral elements.Beside the traditional use,A.annua has also shown antioxidant,anti-inflammatory and antibacterial activities and nutritional characteristics,which make it a promising botanical feed additive for farm animals.However,the herb is generally used as antiparasitic in animals,especially anticoccidiosis in poltry.Enzymatically treated A.annua L.(EA)was the entire herb of A.annua treated by cellulases and pectinase.Hydrolyzing and degrading or decomposing the cell wall of plant by enzymatic treatment facilitate the release of intracellular contents and make full use of bioactive compounds of the plant.Heat stress induced by high ambient temperature is a common stressor in farm animals.Dietary supplementation with phytogenic feed additives is a satisfactorily feasible approach that has been developed to alleviate the detrimental effects of heat stress in animals.However,the antioxidant capacity of EA and the effects of EA in alleviating heat stress remain to be investigated.Therefore,this study was conducted to(1)evaluate the antioxidant capacity of EA in vitro,as well as its application in diet on growth performance and antioxidant capacity of broilers,(2)evaluate the effects of varying levels of EA on growth performance,meat quality,and antioxidant capacity of breast and thigh muscles in broilers,(3)clarify the effects of dietary EA supplementation on growth performance,blood biochemistry,hepatic histology and structure,and oxidative status of broilers exposed to heat stress,and(4)investigate the potential mechanism of EA in mitigating the adverse effects of heat stress in breast muscle.1 Antioxidant capacity of enzymatically treated Artemisia annua L.in vitro and in broilersThe antioxidant capacity of EA was investigated both in vitro and in broilers in the current study.Total phenolic and flavonoid contents were determined by Folin-Ciocalteu and aluminium chloride methods,respectively.Antioxidant capacities were evaluated by scavenging activities against 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)diammonium salt(ABTS)free radical and 2,2-diphenyl-1-picrylhydrazyl(DPPH)free radical,ferric reducing antioxidant power(FRAP)test and reducing power(RP)test.The total phenolic and flavonoid contents of EA were 4.77 mg GAE/g and 1.68 mg RE/g,respectively.The scavenging activities of ABTS·+ and DPPH· of EA were 36.15 ?mol TE/g and 18.17%,respectively.The FRAP and RP of EA were 20.62 ?mol FeSO4/g and 0.20(absorbance at 700nm),respectively.The effects EA on growth performance and antioxidant capacity of broilers were investigated by using 128 one-d-old broilers.Birds were randomly allotted into two groups with eight replicates of eight birds each,and received basal diet and basal diet supplemented with 1 g/kg EA until 42 d of age,respectively.(1)Compared to control,EA increased ADG in 1-21 d,22-42 d,and 1-42 d(P<0.05),and ADFI in 22-42 d and 1-42 d(P<0.05).(2)Dietary EA supplementation increased activities of total superoxide dismutase(T-SOD),glutathione peroxidase(GSH-Px),and catalase(CAT)in serum at 21 and 42 d(P<0.05),and decreased malondialdehyde(MDA)concentration in serum at 21 and 42 d(P<0.05).(3)Dietary EA supplementation increased activities of T-SOD and GSH-Px in liver at 21 and 42 d(P<0.05),increased CAT activity in liver at 42d(P<0.05),and decreased MDA concentration in liver at 21 and 42 d(P<0.05).In conclusion,EA would be effective in improving growth performance and antioxidant capacity in broilers.2 Evaluation of enzymatically treated Artemisia annua L.on growth performance,meat quality,and oxidative stability of breast and thigh muscles in broilersAn experiment was conducted to evaluate the effects of including EA in broiler diets on growth performance,meat quality,and oxidative stability of breast and thigh muscles.A total of 384 one-d-old Arbor Acres broiler chicks were randomly allotted into six groups with eight replicates of eight birds each.Broilers in the control group were fed basal diet.Broilers in the other five groups were offered basal diet supplemented with 0.50,0.75,1.00,1.25,and 1.50 g/kg EA during the 42-d experiment,respectively.(1)Broilers fed diet supplemented with 1.00 g/kg EA had higher ADG than control group(P<0.05).(2)Diet supplemented with 0.75,1.00,and 1.25 g/kg EA decreased drip loss at 24 h of breast muscle and drip loss at 48 h of thigh muscle(P<0.05).Diet supplemented with 0.75,1.00,1.25,and 1.50 g/kg EA decreased drip loss at 24 h of thigh muscle(P<0.05).Diet supplemented with 1.00,1.25,and 1.50 g/kg EA reduced shearing force of thigh muscle(P<0.05).EA quadratically decreased drip loss at 48 h of breast muscle(P<0.05).Drip loss at 24 h and shearing force of breast muscle were linearly(P<0.05)and quadratically(P<0.05)decreased by EA addition.The drip loss at 24 h and 48 h,cooking loss and shearing force of thigh muscle followed the same fashion.(3)Broilers in the 1.00 g/kg EA group had increased ABTS·+ scavenging capacity of breast muscle(P<0.05).Broilers in the 0.75 and 1.00 g/kg EA groups had increased ABTS·+ and DPPH· scavenging capacities of thigh muscle(P<0.05).The supplementation of EA quadratically increased ABTS·+ and DPPH·scavenging activities in breast and thigh muscles(P<0.05).(4)Dietary EA supplementation reduced average MDA concentrations in breast and thigh muscle samples during 15 d of storage at 4?.Increasing levels of EA linearly and quadratically(P<0.05)decreased the malondialdehyde(MDA)concentrations in breast and thigh muscles at 0,4,8,and 15 d of storage at 4?(except breast muscle at 0 d,quadratic P = 0.004).The results indicated that EA supplementation improved growth performance,meat quality and oxidative stability of breast and thigh muscles in broilers.The inclusion level of 0.75-1.00 g/kg EA in broiler diet was recommended.3 Effects of dietary enzymatically treated Artemisia annua L.supplementation on growth performance,blood biochemistry and liver of broilers exposed to heat stressThe experiment was conducted to evaluate the effects of EA on growth performance and some blood parameters of broilers exposed to heat stress(HS).A total of 32022-day-old Arbor Acres broilers were randomly allotted into five groups with eight replicates of eight birds each.Broilers in the control group were housed at 22 ± 1? and fed the basal diet.Broilers in the HS,HS-EA0.75,HS-EA1.00,and HS-EA1.25 groups were reared under HS(32±1? for 8 h/d and 22 ± 1? for 16 h/d),and fed basal diet supplemented with 0,0.75,1.00,and 1.25 g/kg EA,respectively.The experiment ended at 42 d of age.(1)Dietary 1.00 and 1.25 g/kg EA decreased blood pH and elevated final BW,ADG,and ADFI compared to the HS group(P<0.05).EA linearly and quadratically(P<0.05)increased the BW,ADG,and ADFI of the heat-stressed broilers.(2)Broilers fed EA diets had lower serum concentrations of corticosterone and activities of alanine aminotransferase and aspartate aminotransferase(Linear and Quadratic,P<0.05),and higher serum tri-iodothyronine concentration and tri-iodothyronine/thyroxine(Linear and Quadratic,P<0.05)than the HS group.(3)Compared to the HS group,broilers fed diets supplemented with 1.00 and 1.25 g/kg EA decreased ROM concentration of serum(P<0.05),broilers fed diets supplemented with 0.75,1.00 and 1.25 g/kg EA decreased MDA concentration of serum and ROM,MDA and PC concentrations of liver(P<0.05).Dietary EA supplementation linearly and quadratically reduced the concentrations of ROM,MDA,and PC of serum and liver in heat-stressed broilers(P<0.05).(4)Compared to the HS group,broilers in HS-EA0.75,HS-EA1.00,and HS-EA1.25 g/kg groups had elevated T-SOD activity in serum(P<0.05).Broilers in HS-EA1.00 and HS-EA1.25 groups had increased CAT activity in serum,and GSH concentration and T-SOD,GSH-Px,and CAT activities,and activities to inhibit OH· and O2-· in liver(P<0.05).Broilers in the HS-EA1.00 group had enhanced GSH concentration and activity to inhibit OH· in serum(P<0.05).Dietary EA supplementation linearly and quadratically increased GSH concentration,T-SOD and CAT activities and activities to inhibit OH· and O2-· in serum and liver of heat-stressed broilers(P<0.05),and quadratically increased GSH-Px activity in serum of heat-stressed broilers(P<0.05).(5)Diet supplemented with 0.75,1.00,and 1.25 g/kg EA alleviated the hepatic histology lesion and reduced hepatic cell apoptosis(P<0.05).(6)EA treatment(0.75,1.00,and 1.25 g/kg)down-regulated the mRNA expression levels of heat shock protein 70 and 90,and up-regulated the mRNA levels of nuclear factor erythroid 2-related factor 2,MnSOD and CAT in liver of heat-stressed broilers(P<0.05).The heme oxygenase 1,Cu/ZnSOD,and GSH-Px mRNA levels were increased in the HS-EA1.00 and HS-EA1.25 groups compared to the HS group(P<0.05).(7)In comparison to the HS group,the level of miR-122 was increased and miR-155 was decreased in the 1.00 and 1.25 g/kg EA groups(P<0.05),and the level of miR-200a was increased and miR-144 was decreased in the 0.75,1.00,and 1.25 g/kg EA groups.In conclusion,dietary 0.75-1.25 g/kg EA addition improved growth performance,endocrine,and redox status in heat-stressed broilers,and alleviated the hepatic histology lesion and cell apoptosis induced by HS.4 Dietary enzymatically treated Artemisia annua L.supplementation improves meat quality,antioxidant capacity and energy status of breast muscle in heat-stressed broilersThis study was conducted to evaluate the effects of dietary enzymatically treated Artemisia annua L.(EA)supplementation on meat quality,antioxidant capacity and energy status of breast muscle in heat-stressed broilers.The experimental design was consistent with part 3.(1)The inclusion of EA in diet alleviated the reduction of redness value(1.00 and 1.25 g/kg)and the increase of drip loss at 24 and 48 h(1.00 g/kg)of breast muscle in heat-stressed broilers(P<0.05).EA linearly and quadratically increased pH24 and redness value,and decreased lightness value,drip loss at 24 and 48 h,and cooking loss of breast muscle in heat-stressed broilers(P<0.05).(2)Broilers in HS-EA0.75,HS-EA1.00 and HS-EA1.25 groups had lower MDA concentration than the HS group(Linear and Quadratic,P<0.05).The reduced ROM,improved activities of T-SOD and CAT,FRAP and free radical scavenging abilities of ABTS·+,OH·,and O2-· were observed in HS-EA1.00 and HS-EA1.25 groups compared to the HS group(P<0.05),and respond in a dose dependent manner(Linear and Quadratic,P<0.05).(3)Broilers fed diet supplemented with 0.75-1.25 g/kg EA had higher adenosine triphosphate(ATP)concentration,total adenine nucleotides(TAN)and energy charge(EC)than the HS group(P<0.05).The elevated ATP/adenosine diphosphate(ADP)was observed in HS-EA1.00 and HS-EA1.25 groups compared to the HS group(P<0.05).Increasing concentration of EA linearly and quadratically increased ATP concentration,ATP/ADP,TAN,and EC in breast muscle of heat-stressed broilers(P<0.05).(4)In breast muscle of heat-stressed broilers,dietary 0.75-1.25 g/kg EA supplementation reduced the mRNA level of heat shock protein 70(P<0.05).Dietary 1.00 and 1.25 g/kg EA decreased the mRNA level of heat shock protein 90 whereas increased the mRNA levels of avian uncoupling protein and sirtuin 1,and 1.25 g/kg EA up-regulated the mRNA level of peroxisome proliferator-activated receptor-y coactivator-1?(P<0.05).In conclusion,dietary EA supplementation improved meat quality by increasing antioxidant capacity and energy status in breast muscle of heat-stressed broilers,possibly through altering pertinent mRNA expression levels.Therefore,dietary EA supplementation increased the growth performance,antioxidant capacity and meat quality,alleviated the detrimental effects of heat stress in broilers.The appropriate inclusion level of EAis 0.75-1.00 g/kg based on this study. |
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