Effects of Various Copper Sources and Concentrations on Performance, Skeletal Growth, and Mineral Content of Excreta in Broiler Chickens

Abstract

The experiment was designed to study the effect of supplemental sources and concentrations of copper on the performance and development and mineralization of tibia bones in broiler chickens. 42-day feeding trial using three copper sources i.e., copper sulphate (CuS), copper chloride (CuCl), and copper propionate (CuP) each with four copper levels i.e., 8, 100, 150, and 200 mg/kg was taken. During 4–6 wk of age, the body weight gain was significantly higher at 200 mg Cu/kg diet. There was no significant difference on body weight gain due to interaction between Cu sources and levels. The feed intake during different growth phases did not differ significantly due to either main effect or interaction between copper sources and levels. The feed conversion ratio during 4–6 and 0–6 wk was significantly (P < 0.05) better in CuP supplemented diet (200 mg/kg diet). A total of 72 tibia bones, six per treatment, were collected at the end of experiment. Metabolic trial was conducted to look into mineral retention in broiler chickens on the final three days of the trial (40-42d). 8mg Cu/kg diet with Cu chloride, 100mg Cu/kg diet with Cu propionate, 8mg Cu/kg diet with Cu sulphate, and 8mg/kg diet with Cu propionate supplemented diet resulted in significantly (P < 0.05) increased tibia bone zinc (Zn) levels. Significantly (P < 0.01) lower tibia Zn content was recorded at higher levels of Cu (150 and 200 mg/kg diet). Higher (P < 0.01) tibia Cu content was recorded at 8 mg Cu/kg diet with Cu sulphate treated group. Excreta Zn content was higher (P < 0.01) in Cu sulphate supplemented diet than Cu chloride supplemented diet and lowest excreta Zn content was recorded in Cu propionate supplemented diet. Higher excreta Fe content (P < 0.05) was observed in Cu sulphate and Cu chloride supplemented diet than that recorded in Cu propionate supplemented diet. With the exception of a decrease in the zinc content of the tibia, feeding dietary Cu concentrations up to 200 mg Cu/kg diet, regardless of the different sources, showed no adverse impact on bone morphometry and mineralization parameters.

Introduction

An essential trace element for the maintenance, growth, health, and survival of chickens is copper (Cu). As a co-factor for various metalloenzymes, copper (Cu) is engaged in a wide range of biological activities, including hormone secretion pathways, connective tissue maturation, erythropoiesis, mitochondrial respiration, and immune system defences [1, 2]. In developing broiler chickens, Cu deficiency impairs performance goals and immune processes [3] and causes skeletal system abnormalities. Leg disorder including abnormalities and weakness are continue to be a serious and loss incurring factor for optimum production of broiler chickens. Copper (Cu) is an essential trace minerals required for several physiological functions including bone growth and development in poultry. Deficiency of Cu leads to fragile bone and lameness in broiler chickens. Inadequate supply of dietary Cu causes reduction in tibia bone development and mineralization traits [4, 5]. Removal of trace minerals had a negative effect on bone strength [6].

The National Research Council's (NRC) Poultry Subcommittee determined that broiler requirements for copper were 8 mg/kg feed to prevent deficiency symptoms and sustain growth in the last revision of Nutrient Requirements of Poultry [7]. Cu sources and levels did not produce significant changes in length, width (proximal and distal) [8]. However dietary Cu levels caused considerably larger mid shaft width at 12 mg Cu/kg diet, compared to the other two levels 8 and 16 mg Cu/kg diet. Dietary combination of 40mg Zn/kg and 16mg Cu/kg diet using organic source of mineral were found sufficient to obtain optimum bone morphometry and mineralization traits in broiler chickens [9]. Deficiency of Cu (less than 1ppm) was showed to decrease collagen formation and also lower mineralization. The bio- availability of a mineral from the diet is manifested by the efficiency with which the body utilized and retained the dietary mineral [10]. The retention of minerals influenced by a number of dietary factors, including diet or ingredient type, sources and concentration of minerals and relative proportions of various minerals in the diet [11]. There has been environmental concern of the present society on Cu accumulating in soil and water. The poultry feed industry is still using a large safety margin in feed formulation which causes accumulation of copper in soil and inhibition of normal fermentation process [12, 13].

Considering the importance of Cu as an essential trace mineral for broiler chickens the present study was thus planned to study the effect of feeding different sources and concentrations of Cu on morphometry and mineralization of tibia bone and utilization of minerals in broiler chickens.

Materials And Methods

Experimental Design, Population And Diets

Day-old coloured broiler straight-run chicks (n = 360) were distributed at random on the basis of initial body weight in 36 groups of 10 chicks each reared in battery brooders. Twelve dietary treatments with three dietary Cu sources (copper sulphate- CuS, copper chloride-CuCl and copper propionate-CuP) each at four levels of copper (8, 100, 150 and 200 mg/kg) in following factorial design 3 x 4 was conducted during starting (0–3 wk) and finishing (4–6 wk) growth phases. The ingredients and chemical composition of basal diets for starting and finishing phase are presented in Table 1.

Production Performance

Weekly body weight and feed intake were recorded and feed conversion ratio (unit feed intake/unit body weight gain) was calculated.

Metabolic Trial

During the feeding trial, last three days (40-42days) a metabolic trial were conducted to study the minerals retention/ utilization in broiler chickens. The net feed consumed by each bird in the respective dietary group was recorded and the dropping voided over the same period was collected quantitatively. On the last day, the feeders were removed to determine the net feed intake and faecal trays were removed to collect the faeces in aluminium dishes.

Faecal Excreta

The dropping collected were dried for 4–5 d in oven at 60 ± 5ºc till a constant weight was attained which represented the net dried faecal output. The dried and pooled excreta samples were ground and stored in air tight container for further analysis of different minerals. The experimental diet and excreta samples were analysed for calcium content [14], phosphorus [15], copper, zinc, manganese and iron content were analysed by Atomic absorption spectrophotometer (Varian Spectra AA220 Model).

Bone Mineralization

At the conclusion of the 42-day trial, six birds from each feeding regimen (126 = 72 birds) were slaughtered, and their left tibia bones were extracted and freed of any attached soft tissues. According to Deo et al. [16] bone length and width were measured. Using Vernier callipers, the maximum width of the mid shaft, proximal and distal widths, as well as the overall length of each tibia bone, were measured. Each tibia bone was defatted for 18 hours using the Soxhlet apparatus and petroleum spirit extraction to analyse the bone mineralization. The bones were then oven dried at 100° for 24 hours before being ashed at 650° for 6 hours in a muffle furnace. The tibia ash was measured and expressed as percentage of dry bone weight. The calcium and phosphorus content was estimated by method of Talapatra et al. [14] and AOAC [15], respectively. The bone zinc, copper, manganese and iron content in tibia bone were determined by method of AOAC [15] using AAS (Varian Spectra AA 220 Model).

Results

Production Performance

The body weight gain was noticeably higher at 200mg Cu/kg diet during the first 4–6 weeks of life. Due to the interaction of Cu sources and levels, there were no appreciable differences in body weight gain. Due to neither the main effect of copper sources and levels, nor their interaction, the feed intake during various growth phases did not differ significantly. The feed conversion ratio was significantly (P ≤ 0.05) higher in the CuP supplemented diet (200 mg/kg diet) during the 4–6 and 0–6 wk.

Bone Morphometry

The effect of feeding different sources and concentrations of copper on bone length, width (proximal, mid shaft and distal) are summarized in (Table 2). The various bone morphometry traits such as tibia bone length, width (proximal, mid shaft and distal) did not differed significantly (P ≤ 0.05) due to either main effect or interaction between different sources and concentrations of copper in the diet (Table-2).

Bone Mineralization

The effect of feeding different dietary sources and concentrations of copper on bone mineralization traits are presented in (Table 3).Various bone mineralization traits such as dried bone weight, bone ash, bone calcium, phosphorus and manganese content remained statistically unchanged due to either main effect or interaction between different sources and concentrations of copper in the diet. Significantly (P ≤ 0.05) higher tibia bone zinc (Zn) was recorded at 8mg Cu/kg diet with Cu chloride followed by 100mg Cu/kg diet with Cu propionate, 8mg Cu/kg diet with Cu sulphate and 8mg/kg diet with Cu propionate supplemented diet than those observed in other dietary combinations. Significantly (P ≤ 0.05) higher tibia Zn content was observed in Cu propionate supplemented diet than that recorded with Cu sulphate supplemented diets. However, Cu chloride supplemented diet, the tibia Zn content was found intermediary. Significantly (P ≤ 0.01) lower tibia Zn content was recorded at higher levels of Cu (150 and 200 mg/kg diet) than those recorded at lower levels of Cu (8 and 100 mg/kg diet). Significantly (P ≤ 0.01) higher tibia Cu content was recorded at 8mg Cu/kg diet with Cu sulphate followed by 150mg Cu/kg diet with Cu propionate and 100mg Cu/kg diet with Cu propionate than those recorded in other dietary combinations (Table-4).

Discussion

Present results get strength from the previous finding reported by Kulkarni [27] who reported that Cu sources (copper propionate and copper sulphate) and levels (8, 12 and 16 mg/kg) could not bring any significant change in length and width (proximal, mid shaft, distal) of tibia bone of broiler chicks. Similarly, Zheng et al. [28] reported that measurement evaluating in terms of length, proximal width, mid shaft width and distal width of femurs and tibiae did not reveal substantial benefit to skeletal integrity from above 40 mg Zn/kg supplementation. Iqbal et al. [9] also reported that bone length, proximal width and mid shaft width of bone did not alter statistically due to Cu sources and levels.

Present results get support from earlier observation reported by Hashish et al. [29] who also observed a non-significant effect on tibia bone weight, calcium and phosphorus content of tibia bone by feeding 0 to 200 mg copper/kg diet. In the present finding lower tibia Zn content was recorded at higher levels of Cu (150 and 200mg/kg diet) than those recorded at lower levels of Cu (8 and 100mg/kg diet). The reason may be due to the interaction between Cu and Zn contents in the diet, because excess concentration of dietary copper will decrease the availability of zinc due to antagonistic effect. Thus deposition of zinc in the bone will decrease at higher levels of copper. In the present finding tibia bone Cu content was changed significantly due to copper sources [30].

These results received support from the earlier work reported by Leeson [31] who also reported that when bioavailability of reagent grade Cu sulphate was set as 100% the relative bioavailability of Cu was 111.63% in 14 days and 110.7% in 35 days of broiler chicks. Similarly, organic Cu sources, such as proteinate and amino acid chelate, have been shown to have higher relative bio-availability than that of inorganic mineral sources, such as oxide and sulphate [13]. In contrary to present finding Iqbal et al [9] who reported that tibia bone copper content was varied due to different levels of copper in the diet. These results partially support by earlier finding reported by Ao et al [32] who observed that organic form of mineral sources such as proteinate and amino acid chelate have been shown to have relative bioavailability than that of inorganic mineral sources, such as oxide and sulphate which may influenced the higher iron content in tibia bone in copper propionate supplemented diet. Reeves et al. [33] who observed that copper facilitates the absorption of iron in the body.

The concentrations of calcium (Ca), phosphorus (P), Cu, manganese (Mn), Zn and iron (Fe) in excreta did not differed significantly due to interaction between different Cu sources and concentrations (Table 4). However, as increasing dietary Cu concentration, increased excreta Cu concentration with dose response. Present result is in close agreement with earlier observation reported by Bao et al. [34] who reported that the excretion of Cu increased linearly with increasing intake of the element through the diet. In the present finding excreta Cu content was not change significantly due to copper sources, but numerically less excreta copper content was observed in organic source of copper than inorganic sources of copper, which indicate higher retention of copper in organic source than inorganic source of copper. These result received support from the earlier observation of Lee et al. [35] also reported that excreta Cu content was greatly reduced when organic source of mineral supplemented in broiler diet. Similarly, Shamsudeen et al. [36] reported that a higher retention of Cu in Cu chelate group than its counterpart Cu-inorganic group in broiler chickens. Iqbal et al. [9] also found non-significant effect on excreta calcium and phosphorus content when diet fed variable levels of Cu, Zn and their sources in broiler diets. These results also support present finding. Higher excreta Mn content was recorded in Cu chloride supplemented diet than Cu sulphate supplemented diet and significantly lowest excreta Mn content was recorded in Cu propionate supplemented diet. The lower excreta Mn content found in Cu propionate supplemented diet which suggesting that organic source of Cu had higher retention of minerals than inorganic source copper chloride and copper sulphate. Lee et al. [35] also reported that excreta Cu content was greatly reduced when organic source of mineral supplemented in broiler diet Shamsudeen et al. [36] reported that a higher retention of Cu in Cu chelate group than its counterpart Cu-inorganic group in broiler chickens. Excreta Mn content was recorded higher at 200mg/kg than 150mg/kg diet and lower at 8 and 100mgCu/kg diet. Contrary to the present finding El-Damrawy et al. [37] who reported that manganese retention was unaffected by source and dose of copper in the diet. Excreta Zn content was higher in Cu sulphate supplemented diet than Cu chloride supplemented diet and lowest excreta Zn content was recorded in Cu propionate supplemented diet than those observed in Cu sulphate and Cu chloride supplemented diet. Higher excreta Fe content was observed in Cu sulphate and Cu chloride supplemented diet than that recorded in Cu propionate supplemented diet. The lower excreta Zn and Fe content found in Cu propionate supplemented diet which suggesting that organic source copper propionate had higher retention of Zn and Fe than inorganic sources copper sulphate and copper chloride. El-Damrawy et al. [37] also observed that supplementation of copper propionate had higher Zn and Fe retention than its counterpart copper sulphate supplemented diet.

Therefore, it can be concluded that feeding dietary Cu concentrations up to 200 mg Cu/kg diet, effective in promoting growth, feed conversion efficiency regardless of different sources, and had no negative effects on bone morphometry and mineralization features, with the exception of a decrease in tibia zinc content., However, compared to Cu sulphate and Cu chloride supplemented diets; Cu propionate considerably increased the retention of trace minerals.

Declarations

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