Copper sulfate and cupric citrate supplementation improves growth performance, nutrient utilization, antioxidant capacity and intestinal microbiota of broilers Cupric citrate addition on performance in broilers

doi:10.21203/rs.3.rs-3103766/v1

Download PDF

Research Article

Copper sulfate and cupric citrate supplementation improves growth performance, nutrient utilization, antioxidant capacity and intestinal microbiota of broilers Cupric citrate addition on performance in broilers

https://doi.org/10.21203/rs.3.rs-3103766/v1

This work is licensed under a CC BY 4.0 License

Version 1

posted

You are reading this latest preprint version

This experiment was conducted to study the effects of copper sulfate and cupric citrate on growth performance, nutrient utilization, antioxidant capacity and intestinal microbiota of broilers. A total of 360 one-day-old Ross 308 broilers were randomly divided into 5 groups with 6 replicates in each group and 15 broilers per replicate. Broilers in the control group were fed a basal diet, and animals in other four groups were fed basal diets supplemented with 2 sources (copper sulfate and cupric citrate) and 2 levels (50 and 100 mg/kg dry matter). The experiment lasted for 42 days. The results showed that dietary cupric citrate supplementation increased the average daily gain (P = 0.0313). The average daily feed intake and feed gain ratio, however, were not affected by dietary copper sulfate or cupric citrate (P > 0.10). Additionally, dietary copper sulfate or cupric citrate supplementation increased the digestibility of crude protein (P = 0.0554) and energy (P = 0.0191). For intestinal microflora, dietary cupric citrate supplementation decreased the concentration of Lactobacillus and Escherichia coli (P < 0.05) in the ileal digesta or cecal digesta. In addition, dietary Cu supplementation increased the pH in duodenum (P = 0.0008) and jejunum (P = 0.0589). The activities of serum Cu-Zn superoxide dismutase (P = 0.0899), and ceruloplasmin (P = 0.0269) were increased by Cu addition. The present study demonstrated cupric citrate fed to broilers has a positive effect on growth and nutrient utilization. Our results also show that moderately high Cu in the diet increases the pH in duodenum and jejunum, and reduced the concentration of Lactobacillus and Escherichia coli in the ileal digesta or cecal digesta.

cupric citrate

intestinal microbiota

broiler

lipid profiles

antioxidant defenses

immune

Copper is a trace element widely distributed in organisms. It is involved in the composition of plasma ceruloplasmin, copper zinc superoxide dismutase (Cu/Zn SOD), cytochrome c oxidase, monoamine oxidase and other enzymes. Copper citrate can be used as a new copper source to replace copper sulfate in feed because of its reducing the destruction of nutrients and high biological potency. Since Braude [1] found that adding high copper (100-250mg / kg) to feed can improve the growth performance of piglets in 1945, many studies have confirmed that feeding high copper can improve the average daily gain, average daily feed intake and feed conversion efficiency of animals, reduce the pH value of intestinal chyme and improve the activities of proteinase and phospholipase[2, 3].

Copper has a wide range of bactericidal spectrum and is effective for bacteria, molds and fungi. Dietary copper supplementation could reduce the infection of broilers with Eimeria [4] and and inhibit the growth of Escherichia coli (E. coli). [5]. Peng et al. [6] found that adding 60 mg / kg copper citrate to piglet diet can improve growth performance, reduce copper residue in the body, improve antioxidant function, reduce the number of cecal E. coli and increase the number of Lactobacillus. Yan et al. [7] found that adding 20 mg / kg copper citrate to piglet diet can increase the expression of bone marrow antimicrobial peptides and significantly reduce the diarrhea rate of piglets.

Citric acid is an important bioactive compound and a chelating ligand with strong coordination ability [8]. Citric acid chelates have the advantages of high bioavailability, compared with other organic acid chelates, the price is cheaper, more in line with the market demand [9, 10]. Copper citrate is a new feed additive approved by Ministry of Agriculture and Rural Affairs of China in 2019. The research on copper citrate in diet mainly focuses on piglets, while the research on the application of copper citrate in broiler diet is rarely reported. To this end, this experiment used Ross 308 broilers as the object to study the effects of adding copper citrate on growth performance, nutrient utilization, antioxidant capacity and intestinal microbiota of broilers. It aims to provide a basis for the scientific application of copper citrate in broiler production.

Ethics committee approval was received from Anhui Science and Technology University Ethics Committee with decision number 2021012 on 01/03/2021. The experiment was conducted in accordance with guidelines approved by the Animal Health and Care Committee of Anhui Science and Technology University.

Animals, diets, and management

Three hundred and sixty 1-day-old healthy broiler chicks (Ross 308) were allotted to 5 groups with 6 replicates in each group and 12 broilers in each replicate. Dietary treatments were as follows: (1) basal diet without supplemental Cu (control); (2)basal diet + 50 mg Cu/kg dry matter (DM) as copper sulphate (CS-50); (3) basal diet + 100 mg Cu/kg DM as copper sulphate (CS-100); (4) basal diet + 50 mg Cu/kg DM as copper citrate (CC-50) and (5) basal diet + 100 mg Cu/kg DM as copper citrate (CC-100). The experiment lasted for 42 days, and diets were formulated in two stages (1 ~ 21 days old and 22 ~ 42 days old). Ingredients and chemical composition of the basal diets are presented in Table 1. The final Cu content of the diets are presented in Table 2.

Table 1

Ingredient and chemical composition of basal diet
Item	Starter	Finisher
Ingredient, %
Corn	55.2	57.6
Soybean meal (48% CP)	37.0	34.0
Soybean oil	3.0	4
Dicalcium phosphate	2.0	1.4
Ground limestone	0.8	0.9
Salt	0.4	0.4
DL–Methionine	0.3	0.3
L-Lysine	0.3	0.4
Premix¹	1	1
Analyzed composition
Metabolisable energy² [MJ/kg DM]	12.40	12.76
Dry matter [g/kg]	912.1	903.4
Crude protein [g/kg DM]	220.3	207.8
Contents of animo acid [ %]
Lysine	1.53	1.55
Methionine	0.63	0.62
Methionine + cyseine	0.99	0.95
Contents of mineral elements [mg/kg]
Calcium	10,016	8,641
Available phosphorus	5,476	4,239
Copper	7.81	7.03
¹Provided per kilogram of diet: 11,000 IU vitamin A; 1,800 IU vitamin D₃; 32 mg vitamin E; 0.5 mg vitamin K₃; 2.5 mg vitamin B₁; 8 mg riboflavin; 4.5 mg vitamin B₆; 26 mg vitamin B₁₂; 65 mg niacin; 1.5 mg folic acid; 0.2 mg biotin; and 18 mg pantothenic acid; 60 mg Zn (as ZnSO₄); 95 mg Mn (as MnO₂); 65 mg Fe (as FeSO₄∙7H₂O); 0.7 mg I (as KI); and 0.2 mg Se (as Na₂SeO₃∙5H₂O).
²Calculated values, others were analyzed values based on dry matte samples.

Table 2

The final Cu content of the diets [mg/kg]
Items	Additive Cu does	Actually Cu does (Starter)	Actually Cu does (Finisher)
Control	0	7.81	7.03
CS-50	50	57.74	57.12
CS-100	100	107.32	107.57
CC-50	50	58.22	57.12
CC-50	100	108.25	107.69

The single-layer cage (200 cm long × 100 cm wide × 40 cm high, with concrete floors of 0.167 m²/bird) was adopted in the experiment. The chicken house and related appliances were cleaned and disinfected before the experiment. The temperature of the chicken house was controlled by human industrial temperature control three days before entering the chicken house. Diets and water (less than 0.01 mg/L Cu by analysis) were available ad libitum.

Sample collections

On the d 1, d 21 and d 42, the weight was measured (stop feeding for 12 hours before weighing and drink freely), and the feed intake was recorded at the same time. Average daily gain (ADG), average daily feed intake (ADFI) and the feed/gain ratio (F/G) were calculated based on the recorded data.

On d 42, 60 birds (2 chicks per replicate) were randomly selected and weighed after feed deprivation for 12 h. Blood samples were collected from the wing vein and centrifuged at 3,500×g for 10 min. Serum was separated and stored at − 80°C for further analyses.

After blood sampling, birds were killed by cervical dislocation immediately and the pectorals and liver were excised in situ. The pH of the digesta in the jejunum and ceca was subsequently measured with a calibrated digital pH meter with a glass-tipped probe (model IQ120, IQ Scientific Instruments Inc., Carlsbad, CA). Two independent pH readings were taken in situ in each location along the digestive tract. The cecal samples and the content samples of the ileal (Meckel’s diverticulum up to 40 mm above the ileo-cecal junc-tion) digesta were aseptically collected and placed on ice for transportation to the laboratory, where fresh cecal samples were diluted 10-fold by weight in buffered peptone wate and homogenized using a stomacher. Viable counts of bacteria in the samples were then determined by plating serial 10-fold dilutions (in 10 g/L peptone solution) onto MacConkey agar, Lactobacilli MRS agar, and Bifidobacterium agar plates (Beijing Luqiao technology Limited by Share Ltd) to verify the E. coli i, Lactobacillus, and Bifidobacterium, respectively. Agar plates were incubated at 37°C for 36 or 48 h, after which bacterial colonies were counted. Concentration of microflora was finally expressed as log10 colony-forming units per gram of intestinal content.

At the 39th to 42nd day of the experiment, the feed intake was recorded and all excreta was collected for 4 consecutive days. Excreta were collected from plastic trays placed under the cages 6 times per day (06:00, 9:00, 12:00, 15:00, 18:00 and 21:00 h) and stored at − 20℃. Finally, feces from each cage were collected, mixed and weighed. Fecal samples were dried in an oven at 65°C for 48 h. Feed and excreta samples were ground through a 0.45 mm screen and stored for further analysis.

Chemical analysis

The nutrient contents of feed were analyzed by the methods of the Association of Official Analytical Chemists (AOAC)[11]. Copper concentrations of feed, tissue and plasma were analyzed by Flame Atomic Absorption Spectroscopy (Shimadzu Scientific Instruments, Kyoto, Japan; Method 999.10 or method 985.01; AOAC, 2005). The activities of serum ceruloplasmin were determined by using Cu-Zn Superoxide Dismutase (SOD) typed assay kit (Hydroxylamine method, Nanjing Jiancheng Bioengineering Institute, Nanjing, P. R. China). The activities of serum Cu-Zn SOD were determined by using a ceruloplasmin Enzyme Linked Immuno Sorbent Assay (ELISA) kit (Shenzhen Zike Biotechnology Co., Ltd).

Statistical analysis

All data were analyzed as a 2 × 2 + 1 factorial experiment based on a completely randomized design using the General Linear Model procedure of the SAS software (Statistical Analysis System, Version 9.13, 2002). Data were analyzed as repeated measures with a model containing source, level, and source × level. Means were compared using Duncan’s multiple range test and P < 0.05 was considered as the significant level, and P > 0.05 and < 0.10 was considered a trend.

Growth performance

The effect of copper sulfate and cupric citrate on growth performance of broilers is found in Table 3. Copper sulfate or copper citrate at 50 mg/kg Cu or 100 mg/kg Cu of the diet had no effect on ADFI of broiler in any period. However, the ADG was increased by Cu supplementation in the periods of 1 to 21 d (P = 0.0313) and the overall period of 1 to 42 d (P = 0.0812). Broilers in the CC-100 groups had higher ADG than those in the control and CS-50 group (P < 0.05). Neither copper sulfate nor cupric citrate affected F/G in any period. Nevertheless, the lowest F/G was seen in the CC-100 group.

Table 3

Effect of copper sulfate and cupric citrate on growth performance of broilers
Items	Treatment					SEM	P-values
Items	Control	CS-50	CS-100	CC-50	CC-100	SEM	Treat	Sources	Level	Sources × Level
Day 1 to 21
ADG, g/d	36.40^c	36.67^bc	38.23^ab	38.15^ab	38.56^a	0.51	0.0313	0.0260	0.0900	0.3155
ADFI, g/d	49.72	49.33	50.51	49.83	51.32	0.90	0.6399	0.7164	0.1854	0.8801
F/G	1.37	1.34	1.32	1.31	1.33	0.02	0.4516	0.2740	0.9542	0.3109
Day 22 to 42
ADG, g/d	85.47	86.73	87.25	89.34	89.45	1.40	0.3110	0.1029	0.8377	0.8940
ADFI, g/d	170.82	169.55	171.03	175.14	171.70	2.89	0.7774	0.5941	0.7601	0.4431
F/G	2.00	1.96	1.96	1.96	1.92	0.03	0.6237	0.4020	0.6043	0.4925
Day 1 to 42
ADG, g/d	60.93^b	61.70^ab	62.74^ab	63.75^a	64.01^a	0.78	0.0812	0.0245	0.4540	0.6534
ADFI, g/d	110.27	109.44	110.77	112.48	111.51	1.62	0.7830	0.5314	0.9193	0.5193
F/G	1.81	1.77	1.77	1.77	1.74	0.02	0.4389	0.1989	0.5469	0.7819
^a−bMeans in a row without common superscripts differ (P < 0.05).
CS copper sulfate, CC cupric citrate, ADG average daily gain, ADFI average daily feed intake, F/G feed gain ratio.

Nutrient Utilization

The effect of copper sulfate and cupric citrate on apparent nutrients digestibility of broilers is found in Table 4. Neither copper sulfate nor cupric citrate affected the apparent digestibility of DM, Ca, or P. However, dietary Cu supplementation increased the digestibility of CP (P = 0.0554) and energy (P = 0.0191). Broilers supplemented copper (as copper citrate) with 100 mg/kg had greater (P < 0.05) CP digestibility than those receiving 0 or 50 mg/kg copper (as copper sulfate). Copper sulfate or copper citrate at 50 mg/kg Cu or 100 mg/kg Cu of the diet had higher (P < 0.05) energy digestibility than those in the control group.

Table 4

Effect of copper sulfate and cupric citrate on apparent nutrients digestibility in broilers
Items	Treatment					SEM	P-values
Items	Control	CS-50	CS-100	CC-50	CC-100	SEM	Treat	Sources	Level	Sources × Level
DM, %	73.27	73.84	75.28	74.66	75.74	0.95	0.4567	0.3371	0.2381	0.8642
Energy, %	75.34^b	78.22^a	79.01^a	79.20^a	79.91^a	0.86	0.0191	0.0042	0.4322	0.9627
CP, %	50.99^b	51.44^b	53.29^ab	52.61^ab	53.91^a	0.69	0.0554	0.0641	0.0459	0.7180
Ca, %	37.98	38.45	38.95	37.66	38.42	0.82	0.8744	0.7114	0.4890	0.8859
P, %	26.16	26.48	26.50	26.07	26.95	0.67	0.9252	0.9196	0.5456	0.5656
^a−bMeans in a row without common superscripts differ (P < 0.05).
CS copper sulfate, CC cupric citrate, DM dry matter, CP crude protein, Ca calcium, P phosphorus.

Intestinal and Excreta Microflora

The effect of copper sulfate and cupric citrate on intestinal microflora of broilers is found in Table 5. Adding copper sulfate and cupric citrate decreased (P < 0.05) the concentration of E. coli in ileum and cecum. Similarly, the concentration of Lactobacillus also showed the same trend. In the ileal digesta, copper sulfate or copper citrate at 100 mg/kg Cu of the diet had lower (P < 0.05) the concentration of Lactobacillus than those in the control group, and broilers in the CC-100 groups had lower (P < 0.05) the concentration of E. coli than those in the CS-50, CC-50 and control group. In the cecal digesta, broilers in the CC-100, CC-50, CS-100 groups had lower (P < 0.05) the concentration of Lactobacillus and E. coli than those in the CS-50 and control group.

Table 5

Effect of copper sulfate and cupric citrate on on intestinal microflora in broilers
Items	Treatment					SEM	P-values
Items	Control	CS-50	CS-100	CC-50	CC-100	SEM	Treat	Sources	Level	Sources × Level
Ileum, log10 cfu/g
Lactobacillus spp.	7.35^a	7.05^ab	6.43^b	6.93^ab	6.55^b	0.23	0.0892	0.1095	0.0552	0.6377
Escherichia coli	5.39^a	4.72^b	4.36^bc	4.56^b	3.98^c	0.16	0.0001	0.0001	0.0126	0.5267
Cecum, log10 cfu/g
Lactobacillus spp.	8.40^a	7.94^a	7.27^b	7.28^b	6.83^b	0.20	0.0002	0.0001	0.0163	0.6262
Escherichia coli	7.31^a	6.86^a	5.86^b	6.09^b	5.36^c	0.16	0.0001	0.0001	0.0001	0.4330
^a−cMeans in a row without common superscripts differ (P < 0.05).
CS copper sulfate, CC cupric citrate.

Table 6

Effect of copper sulfate and cupric citrate on on gastrointestinal pH in broilers
Items	Treatment					SEM	P-values
Items	Control	CS-50	CS-100	CC-50	CC-100	SEM	Treat	Sources	Level	Sources × Level
Glandular stomach	2.96	2.92	2.92	2.99	2.92	0.04	0.7499	0.6996	0.4035	0.4973
Muscular stomach	3.78	3.75	3.73	3.82	3.78	0.05	0.8120	0.5244	0.6503	0.8545
Duodenum	5.86^c	6.02^bc	6.19^ab	6.09^ab	6.25^a	0.05	0.0008	0.0007	0.0112	0.9208
Jejunum	6.03^b	6.06^b	6.18^ab	6.13^ab	6.28^a	0.06	0.0589	0.0765	0.0393	0.8279
Ileum	6.00	6.10	5.92	6.09	5.94	0.09	0.5547	0.9837	0.0936	0.9205
Cecum	7.10	7.08	6.93	7.13	6.97	0.09	0.5108	0.7107	0.1147	0.9157
^a−cMeans in a row without common superscripts differ (P < 0.05).
CS copper sulfate, CC cupric citrate.

Gastrointestinal pH

Neither copper sulfate and cupric citrate affected pH of glandular stomach, muscular stomach, ileum and cecum. However, dietary Cu supplementation increased the pH in duodenum (P = 0.0008) and jejunum (P = 0.0589). In the duodenum and jejunum, broilers in the CC-100 groups had higher (P < 0.05) the pH than those in the CS-50 and control group.

Antioxidant defenses

Table 7 presents the influence of dietary copper sulfate and cupric citrate on serum antioxidant defenses parameters in broilers. Neither Cu-Zn SOD (P = 0.0899) nor ceruloplasmin (P = 0.0269) were increased by Cu addition. Broilers in the CC-100 groups had higher (P < 0.05) the activities of serum Cu-Zn SOD than those in the control group. Broilers in the CC-100, CC-50, CS-100 groups had higher (P < 0.05) ceruloplasmin concentration than those in the control group.

Table 7

Effect of copper sulfate and cupric citrate on on antioxidant defenses in broilers
Items	Treatment					SEM	P-values
Items	Control	CS-50	CS-100	CC-50	CC-100	SEM	Treat	Sources	Level	Sources × Level
Ceruloplasmin (U/L)	1.93^b	2.00^ab	2.07^a	2.05^a	2.09^a	0.03	0.0269	0.0142	0.1126	0.5794
Cu/Zn-SOD (U/mL)	90.75^b	95.98^ab	98.63^ab	103.29^ab	106.69^a	3.77	0.0899	0.0254	0.4708	0.9273
^a−bMeans in a row without common superscripts differ (P < 0.05).
CS copper sulfate, CC cupric citrate, Cu/Zn SOD Cu-Zn superoxide dismutase,.

Many studies have shown that the effect of adding chelated copper to feed is significantly better than that of adding copper sulfate [12, 13]. The results showed that there were no significant differences in ADFI, ADG and F/G of the diets supplemented with copper citrate and copper sulfate. However, dietary supplemented with 50 mg/kg or 100 mg/kg copper citrate has a higher ADG, indicating that copper citrate had a certain effect on promoting the growth of broilers. The increase of feed intake was an important reason that copper citrate promoted the growth of broilers. However, the mechanism of copper citrate induced increase in feed intake of broilers is still unknown and needs to be further studied. The results showed that adding 50 mg / kg copper citrate in the diet could improve the growth performance of broilers, and there was no difference in the growth performance of broilers compared with adding 100 mg / kg copper sulfate. This provides a new idea for adding low-dose copper citrate instead of high-dose copper sulfate to provide copper for broilers in practical production.

Adding copper to feed can improve the activity of enzymes, thus improving the nutrient digestion efficiency [14, 15]. Kirchgessner et al. [16] found that appropriate copper ions can activate pepsin activity and promote protein absorption. Luo, Dove [17] showed that high copper can improve the activities of lipase and phospholipase A in the small intestine, thus improving the absorption of essential fatty acids. Our experiment found that dietary copper supplementation had no significant difference in the digestibility of calcium and phosphorus, while the digestibility of dry matter, energy and crude protein were significantly increased. Wu et al. [18] proved that adding copper to mink diet can improve the activation of fat digestive enzymes and pepsin, thus improving the digestibility of protein in the diet. High dose of copper added to the diet can significantly increase the copper content in the liver, and at the same time increase the excretion of excess copper in the liver [19]. The main way of copper excretion in the body is through bile, resulting in increased bile secretion [20, 21]. The pH of bile is weakly alkaline, and its main function is to promote fat digestion, such as flower fat [22]. This may well explain the rise in jejunum pH and promote fat absorption.

Jejunum is the main place for the digestion and absorption of nutrients in broilers, and its pH has an important effect on the activity of digestive enzymes[23]. The optimum pH of trypsin is about 8 [24], and the pH of intestinal tract near duodenum is almost neutral [25]. No relevant studies have been found on the effect of copper on pH of intestinal contents. We found that dietary supplementation of 50 mg/kg or 100 mg/kg copper, whether copper sulfate or citrate, slightly increased pH of jejunum and duodenum contents in broilers. This may be why copper promotes growth and nutrient utilization in animals.

Peng et al. [6] showed that copper citrate could reduce cecal E. coli and increase lactobacillus, thus reducing diarrhea rate and mortality rate of animals. The results showed that dietary copper citrate had a significant inhibitory effect on E. coli in ileum and cecum of broilers, and the sensitivity of E. coli to copper citrate increased with the increase of copper citrate concentration. Warnes et al. [26] showed that copper substantially reduces bacterial growth and limits bacterial infectious behavior. Tan et al. [27] found that copper inhibits iron-mediated DNA oxidative damage in E. coli. In addition, copper can inhibit the growth of wild-type and mutant E. coli, and the addition of branched chain amino acids restored the growth, indicating that copper hinders their biosynthesis [28]. High concentrations of copper significantly increased the number of Dehalobacterium, Coprococcus, and Spirochaetales in the rectum, while the number of Salinicoccus, Bacillales, Staphylococcus, and Lactobacillales decreased dramatically, interrupting the dynamic balance of the microbiota [29]. The precise mechanism of how copper kills microorganisms may be controlled by many factors, and there are different mechanisms for different microorganisms, which need to be further studied.

Copper is an important component of ceruloplasmin and Cu-Zn superoxide dismutase, which are important antioxidant enzymes in animals and play an important role in maintaining animal health [30]. Ceruloplasmin inhibits the free radical production of iron ions through its ferrous oxidase activity [31]. Cu-Zn superoxide dismutase can inhibit the production of superoxide anion free radicals and protect the structure and function of cell membrane [32]. Many studies have shown that high copper diet can significantly increase the activities of ceruloplasmin and cu-Zn superoxide dismutase in animal serum [33, 34]. The results of this study showed that dietary supplementation of copper citrate or copper sulfate can increase the activities of ceruloplasmin and copper zinc superoxide dismutase in serum of broilers.

The result of this feeding trial demonstrated that cupric citrate fed to broilers has a positive effect on growth and nutrient utilization. Additionally, this study also indicated that moderately high Cu in the diet increases the pH in duodenum and jejunum, and reduced the concentration of Lactobacillus and Escherichia coli in the ileal digesta or cecal digesta of broiler chickens.

Cu-Zn SOD Copper zinc superoxide dismutase

ADG Average daily gain

ADFI Average daily feed intake

F/G Feed/gain

DM Dry matter

CP Crude protein

Ca Calcium

P Phosphorus

Funding

The funding for this study was from Anhui Provincial Natural Science Foundation (2108085MC114), Key research and development Program of Anhui Province (202004a06020050), Major project of Shandong Agricultural Improved Varieties Project (2019LZGC019), High-level Talents Introduction Project of Anhui Institute of Science and Technology (DKYJ201701).

Contributions

All the authors wrote the manuscript and revised the manuscript, reviewed, and agreed on the final manuscript before submission. Mingxia Zhu, Qingkui Jiang contributed to the conception of the study. Xuezhuang Wu，Zhentao Lu and Yunting Zhang performed the experiment. Xuezhuang Wu， Aiyou Wen performed the data analyses and wrote the manuscript. Yahao Zhou, Qingkui Jiang helped perform the analysis with constructive discussions.

Data Availability

Not applicable

Ethics approval and consent to participate

The protocol for the present experiment was approved by the Animal Care Committee of the Anhui Science and Technology University (2021012).

Competing interests

The authors declare no competing interests.

Braude R (1945) Some observations on the need for copper in the diet of fattening pigs. J Agricultural Sci 35(3):163–167. 10.1017/S0021859600049170
Hedemann MS, Jensen BB, Poulsen HD (2006) Influence of dietary zinc and copper on digestive enzyme activity and intestinal morphology in weaned pigs. J Anim Sci 84(12):3310–3320. 10.2527/jas.2005-701
Wu X, Dai S, Hua J, Hu H, Wang S, Wen A (2019) Influence of Dietary Copper Methionine Concentrations on Growth Performance, Digestibility of Nutrients, Serum Lipid Profiles, and Immune Defenses in Broilers. Biol Trace Elem Res 191(1):199–206. 10.1007/s12011-018-1594-5
Santos T, Teng P-Y, Yadav S, Castro F, Koch R, Craig S, Chen C, Fuller A, Pazdro R, Sartori J, Kim WK (2020) Effects of Inorganic Zn and Cu Supplementation on Gut Health in Broiler Chickens Challenged With Eimeria spp. Front Veterinary Sci 7. 10.3389/fvets.2020.00230
Danilova TA, Danilina GA, Adzhieva AA, Vostrova EI, Zhukhovitskii VG, Cheknev SB (2020) Inhibitory Effect of Copper and Zinc Ions on the Growth of Streptococcus pyogenes and Escherichia coli Biofilms. Bull Exp Biol Med 169(5):648–652. 10.1007/s10517-020-04946-y
Peng CC, Yan JY, Dong B, Zhu L, Tian YY, Gong LM (2017) Effects of graded levels of cupric citrate on growth performance, antioxidant status, serum lipid metabolites and immunity, and tissue residues of trace elements in weaned pigs. Asian-Australas J Anim Sci 30(4):538–545. 10.5713/ajas.16.0182
Yan J, Zhang C, Tang L, Kuang S (2015) Effect of Dietary Copper Sources and Concentrations on Serum Lysozyme Concentration and Protegrin-1 Gene Expression in Weaning Piglets. Italian J Anim Sci 14(3):3709. 10.4081/ijas.2015.3709
Fiume MM, Heldreth BA, Bergfeld WF, Belsito DV, Hill RA, Klaassen CD, Liebler DC, Marks JG Jr, Shank RC, Slaga TJ, Snyder PW, Andersen FA (2014) Safety Assessment of Citric Acid, Inorganic Citrate Salts, and Alkyl Citrate Esters as Used in Cosmetics. Int J Toxicol 33(2 suppl):16s–46s. 10.1177/1091581814526891
Lu P-J, Hu W-W, Chen T-S, Chern J-M (2010) Adsorption of copper–citrate complexes on chitosan: Equilibrium modeling. Bioresour Technol 101(4):1127–1134. https://doi.org/10.1016/j.biortech.2009.09.055
Field TB, McCourt JL, McBryde WAE (1974) Composition and Stability of Iron and Copper Citrate Complexes in Aqueous Solution. Can J Chem 52(17):3119–3124. 10.1139/v74-458
AOAC (2005) Official methods of analysis, 18th edn. edn. Assoc. Anal. Chem., Arlington, VA
Wu X, Zhu M, Jiang Q, Wang L (2020) Effects of Copper Sources and Levels on Lipid Profiles, Immune Parameters, Antioxidant Defenses, and Trace Element Residues in Broilers. Biol Trace Elem Res 194(1):251–258. 10.1007/s12011-019-01753-z
Wen A, Dai S, Wu X, Cai Z (2019) Copper bioavailability, mineral utilization, and lipid metabolism in broilers. Czech J Anim Sci 64(12):483–490. doi.org/10.17221/210/2019-CJAS
Shang X, Wang C, Zhang G, Liu Q (2020) Effects of soybean oil and dietary copper levels on nutrient digestion, ruminal fermentation, enzyme activity, microflora and microbial protein synthesis in dairy bulls. 74(4):257–270. 10.1080/1745039x.2019.1679562
Olivares M, Uauy R (1996) Copper as an essential nutrient. Am J Clin Nutr 63(5):791S–796S
Kirchgessner M, Beyer MG, Steinhart H (1976) Activation of pepsin (EC 3.4.4.1) by heavy-metal ions including a contribution to the mode of action of copper sulphate in pig nutrition. Br J Nutr 36(1):15–22
Luo XG, Dove CR (1996) Effect of dietary copper and fat on nutrient utilization, digestive enzyme activities, and tissue mineral levels in weanling pigs. J Anim Sci 74(8):1888–1896
Wu X, Yang Y, Liu Z, Gao X, Yang F, Yang P, Xing X (2018) Effects of dietary copper level on serum lipid metabolism parameters,blood parameters,intestinal digestive enzyme activities and bile trace element contents of minks during winter fur-growing period. Chin J Anim Nutr 30(4):1406–1414. 10.3969/j.issn.1006-267x.2018.04.023
Anastassakis K (2022) Copper (Cu). In: Anastassakis K (ed) Androgenetic Alopecia From A to Z: Vol. 2 Drugs, Herbs, Nutrition and Supplements. Springer International Publishing, Cham, pp 399–404. doi:10.1007/978-3-031-08057-9_46
Kardos J, Héja L, Simon Á, Jablonkai I, Kovács R, Jemnitz K (2018) Copper signalling: causes and consequences. Cell Communication and Signaling 16(1):71. 10.1186/s12964-018-0277-3
Armstrong TA, Spears JW, Van HE, Engle TE, Wright CL (2000) Effect of copper source (cupric citrate vs cupric sulfate) and level on growth performance and copper metabolism in pigs. Asian Australasian Journal of Animal Sciences 13(8):1154–1161. org/10.5713/ajas.2000.1154
Blanco A, Blanco G (2017) Chap. 12 - Digestion - Absorption. In: Blanco A, Blanco G (eds) Medical Biochemistry. Academic Press, pp 251–273. doi:https://doi.org/10.1016/B978-0-12-803550-4.00012-4
Lema I, Araújo JR, Rolhion N, Demignot S (2020) Jejunum: The understudied meeting place of dietary lipids and the microbiota. Biochimie 178:124–136. https://doi.org/10.1016/j.biochi.2020.09.007
Xu Y, Zhang P, Liu X, Wang Z, Li S (2020) Preparation and Irreversible Inhibition Mechanism Insight into a Recombinant Kunitz Trypsin Inhibitor from Glycine max L. Seeds. Appl Biochem Biotechnol 191(3):1207–1222. 10.1007/s12010-020-03254-5
Farner DS (1942) The Hydrogen Ion Concentration in Avian Digestive Tracts. Poult Sci 21(5):445–450. https://doi.org/10.3382/ps.0210445
Warnes SL, Highmore CJ, Keevil CW (2012) Horizontal transfer of antibiotic resistance genes on abiotic touch surfaces: implications for public health. mBio 3(6). 10.1128/mbio.00489-12
Tan G, Cheng Z, Pang Y, Landry AP, Li J, Lu J, Ding H (2014) Copper binding in IscA inhibits iron-sulphur cluster assembly in Escherichia coli. Mol Microbiol 93(4):629–644. 10.1111/mmi.12676
Macomber L, Imlay JA (2009) The iron-sulfur clusters of dehydratases are primary intracellular targets of copper toxicity. Proc Natl Acad Sci U S A 106(20):8344–8349. 10.1073/pnas.0812808106
Cheng S, Mao H, Ruan Y, Wu C, Xu Z, Hu G, Guo X, Zhang C, Cao H, Liu P (2020) Copper Changes Intestinal Microbiota of the Cecum and Rectum in Female Mice by 16S rRNA Gene Sequencing. Biol Trace Elem Res 193(2):445–455. 10.1007/s12011-019-01718-2
Iskandar M, Swist E, Trick KD, Wang B, L'Abbe MR, Bertinato J (2005) Copper chaperone for Cu/Zn superoxide dismutase is a sensitive biomarker of mild copper deficiency induced by moderately high intakes of zinc. Nutr J 4:35. 10.1186/1475-2891-4-35
Zhao K, Wu C, Yao Y, Cao L, Zhang Z, Yuan Y, Wang Y, Pei R, Chen J, Hu X, Zhou Y, Lu M, Chen X (2017) Ceruloplasmin inhibits the production of extracellular hepatitis B virions by targeting its middle surface protein. J Gen Virol 98(6):1410–1421. 10.1099/jgv.0.000794
Furukawa Y, Shintani A, Kokubo T (2021) A dual role of cysteine residues in the maturation of prokaryotic Cu/Zn-superoxide dismutase. Metallomics 13(9):mfab050. 10.1093/mtomcs/mfab050
Liu Z, Wu X, Zhang T, Guo J, Gao X, Yang F, Xing X (2015) Effects of Dietary Copper and Zinc Supplementation on Growth Performance, Tissue Mineral Retention, Antioxidant Status, and Fur Quality in Growing-Furring Blue Foxes (Alopex lagopus). Biol Trace Elem Res 168(2):401–410. 10.1007/s12011-015-0376-6
Wu X, Liu Z, Guo J, Wan C, Zhang T, Cui H, Yang F, Gao X (2015) Influence of dietary zinc and copper on apparent mineral retention and serum biochemical indicators in young male mink (Mustela vison). Biol Trace Elem Res 165(1):59–66. 10.1007/s12011-014-0220-4

No competing interests reported.

Download PDF

Version 1

posted

You are reading this latest preprint version

Copper sulfate and cupric citrate supplementation improves growth performance, nutrient utilization, antioxidant capacity and intestinal microbiota of broilers Cupric citrate addition on performance in broilers

Status:

Version 1

Abstract

Introduction

Materials and Methods

Animals, diets, and management

Sample collections

Chemical analysis

Statistical analysis

Results

Growth performance

Nutrient Utilization

Intestinal and Excreta Microflora

Gastrointestinal pH

Antioxidant defenses

Discussion

Conclusions

Abbreviations

Declarations

References

Additional Declarations

Status:

Version 1