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METABOLISM AND NUTRITION |




* Department of Poultry Science, and
Department of Biomedical Engineering, North Carolina State University, Raleigh 27695;
Department of Health and Biomedical Sciences, Florida Hospital College of Health Sciences, Orlando 32803;
Novus International Inc., 20 Research Park Drive, St. Charles, MO 63304; and # Goldsboro Milling Co., Goldsboro, NC 27532
3 Corresponding author: Peter_Ferket{at}ncsu.edu
| ABSTRACT |
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Key Words: turkey organic trace mineral 25-hydroxycholecalciferol leg problem bone
| INTRODUCTION |
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The increased incidence of lameness in recent years (Vaillancourt et al., 2000; Julian, 2005; V. Felts, unpublished data) may be due in part to the change that has taken place in overall body structure and conformation of fast-growing, high-breast meat yield birds (Abourachid, 1993). Breast muscle yields have increased dramatically in recent years, thereby moving the center of gravity of the bird forward (Abourachid, 1993; Corr et al., 2003a,b; Havenstein et al., 2007). This body conformation characteristic of modern turkeys affects gait patterns (Resch-Magras et al., 1993; Corr et al., 2003b) and imposes considerable stress and strain in femur and tibia bones, thus affecting bone development and increasing the risk of fractures. Femoral spiral fractures and tibia fractures are common pathologies in toms between 15 and 18 wk of age (Vaillancourt et al., 1999, 2000; Julian, 2005). Crespo et al. (1999) reported femoral spiral fractures in turkey breeders and associated them with biomechanical properties (Crespo et al., 2000), microstructure, and trace mineral content of bones (Crespo et al., 2002).
In tom flocks of high average daily gain (>280 g of gain/d), mortality rate associated with leg problems may be as high as 5% of a flock and can occur by exsanguination when the femoral artery is severed by a spiral fracture fragment of the femur or when downer birds are killed by aggressive birds in the flock (Crespo et al., 1999; Julian, 2005). Leg problems of downer birds may include valgus-varus deformations or general leg weakness (Vaillancourt et al., 1999, 2000; Julian, 2005). Leg problems cause economic losses to producers, because birds with these abnormalities exhibit decreased feed intakes, lowered growth performance, increased mortality, and greater rates of downgrades and condemnations during processing (Bennett et al., 2002; Oviedo-Rondón et al., 2006a; Dibner et al., 2007).
Several nutrients can affect skeletal development and lameness in poultry as reviewed by Whitehead (2002) and Oviedo-Rondón et al. (2006b). Vitamin D and its metabolites are perhaps the most extensively studied nutrients with respect to bone development in poultry. Vitamin D3 is important in Ca and P absorption and metabolism, and a deficiency can lead to the failure of mineralization of growing bones, leading to retarded growth, leg weakness, and rickets (Oviedo-Rondón et al., 2006b). Fritts and Waldroup (2003) evaluated supplementation of different concentrations of cholecalciferol (D3) and 25-hydroxycholecalciferol (HyD) in cornsoy diets and found that the incidence and severity of tibia dyschondroplasia were significantly lower among broilers fed diets supplemented with HyD.
Trace minerals often are less recognized as being important for proper bone development. Nonetheless, a sizeable body of literature underscores the importance of Zn, Cu, Mn, and Se among other trace minerals on bone growth and strength (Dibner and Richards, 2006; Oviedo-Rondón et al., 2006b; Dibner et al., 2007). Furthermore, these trace minerals interact with each other and with 1,25-dihydroxycholecalciferol during bone formation, modeling, and remodeling (Beattle and Avenell, 1992).
Trace minerals are most often supplemented to diets as inorganic trace mineral salts (oxides or sulfates), and the mineral requirements for poultry have been established using these inorganic sources (NRC, 1994). However, the bioavailability of inorganic trace minerals can be low and variable due to a variety of nutritional antagonisms, including phytic acid, fiber, Ca, and P (Wedekind et al., 1991; Bremner and Beattie, 1995; Underwood and Suttle, 1999; Tamim and Angel, 2003; Leeson, 2005). Trace mineral (Zn, Cu, and Mn) complexes or chelates with an organic ligand (i.e., amino acid, organic acid, or protein digest) can confer stability of the trace mineral in the upper gastrointestinal system, thereby avoiding mineral losses to antagonists and potentially improving trace mineral bioavailability (Miles and Henry, 2000). Indeed, these organic trace minerals can have greater mineral bioavailability than inorganic trace mineral salts (Paik et al., 1999; Cao et al., 2000; Guo et al., 2001; Leeson, 2005; Predieri et al., 2005; Yan and Waldroup, 2006; Wang et al., 2007). In addition, organic sources of Se (as selenomethionine from high-Se yeast) can be more bioavailable than inorganic Se (Rayman, 2004; Payne and Southern, 2005a,b).
Because of their critical roles in bone development, we hypothesized that the use of more bioavailable forms of vitamin D (i.e., HyD) and trace minerals [i.e., Mintrex (MIN)] may improve early skeletal development and decrease subsequent leg problems without compromising growth performance. The objective of this study was to determine the effect of dietary supplementation of MIN and HyD on the growth performance, incidence of leg abnormalities, and bone fracture resistance of Large White turkey toms.
| MATERIALS AND METHODS |
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Biomechanical Properties of Bone
At 17 wk of age, 2 birds per pen (24 birds per treatment) were killed, and femurs and tibias were dissected and analyzed for biomechanical properties. Fracture resistance of turkey tibias was tested by 4-point bending using an axial servohydraulic load frame (858 Mini Bionix II, MTS Systems Inc., Minneapolis, MN). The tibias were assumed to have a hollow elliptical cross section with a uniform wall thickness as described by Cubo and Casinos (1998, 2000). The diameters of the major axis (medial-lateral direction) and minor axis (frontal-caudal direction) were measured, and bones were loaded with the resulting moment applied about the medial-lateral axis. The 4-point bending tests were carried out with a lower support span of 160 mm. The load was applied through 2 upper supports spaced 76.5 mm apart at a loading rate of 30 mm/min to failure. The cortical wall thickness was measured at 4 points along the cross section on each side of the failure to determine the mean thickness. The total force applied and the displacements of the upper supports were recorded.
The different biomechanical parameters were calculated according to the following formulas:
Area moment of inertia (Ix)

where rmajor = radius of the major axis of the ellipse forming the outer cortical boundary, which is the axis about which the bending takes place; rminor = radius of the minor axis of the ellipse forming the outer cortical boundary; t = average cortical thickness.
The applied moment (M) for 4-point bending:
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where Fmax = maximum applied force; L1 = lower support span; and Lu = upper support span and
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Torsional properties of turkey femurs were tested using an axial-torsion servohydraulic load frame (858 Mini Bionix II, MTS Systems Inc.). Femora were assumed to have a hollow elliptical cross section with inner and outer edges forming similar ellipses. The ends of the femora were potted in epoxy (Bondo, Bondo Corporation, Atlanta, GA) to allow them to be secured into the load frame. Before they were potted, screws were driven into the proximal and distal ends of the bones leaving approximately 10 mm of the screw shaft exposed to help ensure that the ends of bones were tightly held within the epoxy. The distal end was held fixed in the load frame, and the proximal end was rotated in external rotation at a rate of 10°/s until failure. The applied torque and angular rotation were recorded. Cortical wall thicknesses were measured as was done for the tibia, and the radii of the inner ellipses used to model the cross section of the bones were calculated so that the average distance between outer and inner ellipse cortical boundaries matched the averaged measured thickness. The different biomechanical parameters were calculated according to the following formulas:
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where Tmax = maximum torque applied during test and

For similar ellipses: inner elliptical boundary = k x (outer elliptical boundary).
Statistical Analyses
All data were analyzed as a completely randomized block design using 2 x 2 factorial arrangement of dietary MIN and HyD supplementation as main effects. Pen means were used as the experimental units for all variables evaluated. Percentage of mortality and leg abnormality incidence data were transformed to the arcsine square root before analysis, and final data are presented as natural numbers. All measures of statistical significance were based on a probability of P = 0.05 unless otherwise stated. Data were subjected to ANOVA using the GLM procedure of SAS system (SAS Institute, 2003). Means separation was accomplished using Tukeys multiple range tests when a significant F statistic was indicated by ANOVA.
| RESULTS AND DISCUSSION |
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Therefore, the biomechanical properties of tibia and femur bones were evaluated by bending (Table 12
) and torsion (Table 13
) tests, respectively. No significant interaction effects were observed in cortical thickness of bones (t). However, cortical thickness was greater among the MIN-treated birds than the controls (2.21 vs. 1.88 mm, P = 0.001) at site of failure after the 4-point bending tests on the tibiotarsus, but not in torsional tests (P > 0.05) on the femur. The maximum applied moment (M) and the resulting bending stress required for tibia bones to break (
max) in the 4-point bending tests was greater among the MIN supplementation birds than controls (28.6 vs. 25.8 Nm, P < 0.01; and 121.8 vs. 112.6 MPa, P < 0.05, respectively), especially when both MIN and HyD were added to the diet (P = 0.05; Table 12
). The average maximum shear stress (
max) at failure of femoral bones was increased (P < 0.05) by the dietary supplementation of both MIN and HyD (Table 13
). Therefore, dietary MIN and HyD increased the biomechanical strength of tibias and femurs in 17-wk-old turkey toms.
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Significant treatment interaction effects were observed for both mechanical tests. Dietary supplementation of both HyD and MIN resulted in greater values for applied moment and stress at breakage in tibias, and maximum shear of stress at failure in femurs. Synergistic effects of vitamin D and trace minerals have been observed by other researchers. Yamaguchi and Oishi (1989) demonstrated that Zn enhances the ability of 1,25-dihydroxycholecalciferol to increase the activity of bone alkaline phosphatase, an enzyme that is associated with proper bone mineralization. Different sources of vitamin D also enhance the utilization of Cu (Aoyagi and Baker, 1995) and Mn (Biehl et al., 1995).
The effect of organic trace minerals on the torsion test results observed in this study with turkeys are consistent with those observed in broilers (Rucker et al., 1975; Opsahl et al., 1982). None of the diets formulated for this experiment were deficient in any trace mineral, because optimum growth rates were obtained. However, there is evidence that trace mineral requirements for optimal bone development may be greater than for fast growth rates (Dibner and Richards, 2006; Oviedo-Rondón et al., 2006b). The MIN product used in this experiment contained chelated Zn, Mn, and Cu and organic Se. Zinc plays important roles in collagen synthesis and turnover in developing bone, as well as in the regulation of hydroxyapaptite crystallization (Starcher et al., 1980; Wu et al., 1993; Sauer et al., 1997; Nie et al., 1998; Orth, 1999; Wu et al., 2002). Furthermore, gene knockout studies in mice have shown that the Zn-finger transcription factor Gli2 is required for normal endochondral ossification to occur (Miao et al., 2004). Although Zn promotes collagen synthesis and turnover, the collagen and elastin matrices must be stabilized by crosslinking for proper development and to confer tensile strength and elasticity to the bone (Carlton and Henderson, 1964; Opsahl et al., 1982; Rucker et al., 1998; Rath et al., 1999, 2000). This crosslinking is accomplished by the cuproenzyme lysyl oxidase (Opsahl et al., 1982; Rucker et al., 1998). Indeed, bone fragility in response to Cu deficiency in many animal species has been reported as early as the 1940s (Bennett et al., 1948; Tinker and Rucker, 1985), and fractures in femoral bones are frequently associated with decreased serum and bone Cu in several species, including humans (Beattle and Avenell, 1992). Adequate Cu intake is more critical for acquisition of peak bone mass in long bones, whereas adequate and balanced intakes of both Cu and Zn may be more critical for an optimal bone mass in the trabecular bones (Roughead and Lukaski, 2003). Manganese is a cofactor for the glycosyl-transferases that are required for the formation of the mucopolysaccharides that form the hyaline cartilage of immature bone, which are essential for proper development (Fawcett, 1994; Gilbert, 1997; Underwood and Suttle, 1999). Manganese deficiency in poultry species can cause twisting and bending of the tibia, enlargement and malformation of the tibiometatarsal joint, shortening and thickening of long bones, and slipping of the gastrocnemius tendon from its condyles (Underwood and Suttle, 1999). Selenoproteins are involved in bone metabolism and ossification as well (Beattle and Avenell, 1992; Oviedo-Rondón et al., 2006b). In a study of Se deficiency in rats over 2 generations, the deficient rats exhibited multiple signs of abnormal bone development, including a highly significant reduction in bone mineral density (Moreno-Reyes et al., 2001).
Trace minerals are also important to maintain several functions of bone, cartilage, and tendons during daily physical activities (Opsahl et al., 1982; Moreno-Reyes et al., 2001; Speich et al., 2001), especially in fast-growing animals (Beattle and Avenell, 1992). These functions of trace minerals may explain why dietary MIN inclusion decreased the incidence of varus, valgus, or shaky leg abnormalities in the current experiment. The complete etiology of these leg disorders is not well understood, but several authors suggested that trace minerals may decrease their incidence (Beattle and Avenell, 1992; Lilburn, 1994; Julian, 1998).
In conclusion, dietary supplementation of organic trace minerals (Zn, Mn, and Cu) as complexed with methionine hydroxy analog along with Se yeast (Mintrex PSe) can decrease the incidence of leg abnormalities (varus, valgus, and shaky leg) associated with rapid growth. Furthermore, dietary MIN supplementation can improve long bone strength, especially when combined with dietary HyD supplementation. Increased bone strength will likely contribute to fewer broken bones in commercial turkey production. Therefore, the dividends associated with decreased leg abnormalities and a reduction in broken leg bones in the MIN-supplemented birds include improved animal welfare and FCR during the growing and finishing period of turkey toms.
| FOOTNOTES |
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2 The mention of trade names in this publication does not imply endorsement of the products mentioned nor criticism of similar products not mentioned. ![]()
Received for publication May 20, 2008. Accepted for publication September 8, 2008.
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