The efficacy of a standardised product from dried leaves of Solanum glaucophyllum as source of 1,25-dihydroxycholecalciferol for poultry.

1. Chemical characterisation of an extract of Solanum glaucophyllum (SG) leaves affirmed the predominant presence of 1,25-dihydroxycholecalciferol (1,25(OH)2D3) glycosides. The compound 1-(ß-D-glucopyranosyl)-1α,25-dihydroxycholecalciferol was isolated for the first time from a natural source. 2. Vitamin D activity of the extract was confirmed by the calcaemic properties shown in a quail eggshell bioassay. The results suggested a 1,25(OH)2D3 bioavailability of approximately 15%. 3. A broiler feeding experiment replicated in time was carried out with 6 treatments. A basic control diet containing 25 µg cholecalciferol/kg was supplemented with 2.5 and 5 µg free 1,25(OH)2D3/kg, with a product based on dried SG leaves (Panbonis) providing 10 µg of 1,25(OH)2D3-glycosides/kg, with two concentrations of an SG extract providing 8.8 and 37.8 µg of 1,25(OH)2D3-glycosides/kg. 4. Tibia breaking strength and stiffness were numerically greater in all treatment groups with free 1,25(OH)2D3 and with SG products compared to controls, though the overall treatment effects only had probabilities in the range of P = 0.07 to P = 0.1. Values for both characteristics increased progressively, with additions of synthetic 1,25(OH)2D3; values with the dried SG product were similar to those with 5 µg synthetic 1,25(OH)2D3/kg. 5. Plasma calcium was mildly elevated (P < 0.05) in treatment groups. The SG extract treatment containing 37.8 µg 1,25(OH)2D3/kg gave the highest plasma calcium concentration and lowest bodyweight, signs of marginal hypervitaminosis D. Plasma 1,25(OH)2D3 concentrations were in the normal range for all treatments. 6. Tibial dyschondroplasia occurred in only one replicate. The incidences were 31% in controls but considerably lower or zero with all other treatments. 7. Bioavailability of 1,25(OH)2D3 in the SG product seemed to be higher in broiler chickens than in Japanese quails. 8. It is concluded that the inclusion of the dried SG product as a source of vitamin D3 in broiler diets at a dietary concentration of 1 g/kg, providing 10 µg 1,25(OH)2D3/kg, is safe and efficacious.

A QTL for osteoporosis detected in an F2 population derived from White Leghorn chicken lines divergently selected for bone index.

Osteoporosis, resulting from progressive loss of structural bone during the period of egg-laying in hens, is associated with an increased susceptibility to bone breakage. To study the genetic basis of bone strength, an F(2) cross was produced from lines of hens that had been divergently selected for bone index from a commercial pedigreed White Leghorn population. Quantitative trait loci (QTL) affecting the bone index and component traits of the index (tibiotarsal and humeral strength and keel radiographic density) were mapped using phenotypic data from 372 F(2) individuals in 32 F(1) families. Genotypes for 136 microsatellite markers in 27 linkage groups covering approximately 80% of the genome were analysed for association with phenotypes using within-family regression analyses. There was one significant QTL on chromosome 1 for bone index and the component traits of tibiotarsal and humeral breaking strength. Additive effects for tibiotarsal breaking strength represented 34% of the trait standard deviation and 7.6% of the phenotypic variance of the trait. These QTL for bone quality in poultry are directly relevant to commercial populations.

Relationships between genetic, environmental and nutritional factors influencing osteoporosis in laying hens.

1. The effects upon bone quality of feeding limestone in flour or particulate form and housing type (cage or aviary) in lines of hens divergently selected for high (H) or low (L) bone strength over 7 generations were investigated. 2. As in previous generations, highly significant phenotypic differences between lines were observed in all measured bone traits at peak egg production (25 weeks) and towards the end of production (56 weeks) in both cage and aviary systems. 3. At 25 weeks there were no significant effects on bone variables of feeding particulate limestone although a significant reduction in osteoclast number was observed at this age. By 56 weeks osteoclast numbers were further reduced in hens fed particulate limestone and beneficial effects on some bone variables were observed in this treatment group. 4. The genotypic and dietary improvements upon bone quality were independent and additive at both ages. There were very few interactive effects. 5. Hens with the freedom to move in an aviary environment during the laying period had improved bone status compared to caged siblings. Environmental and genotypic effects were additive. 6. There were no effects of line on egg production although H line hens had slightly higher egg production by 56 weeks. Egg numbers were unaffected by diet. Eggshell thickness and strength were unaffected by line but hens fed particulate limestone had thicker- and stronger-shelled eggs over the production period as a whole. 7. We conclude that; (a) genetic selection is extremely effective in improving bone strength and resistance to osteoporosis; (b) allowing hens freedom to exercise can also improve bone strength but may increase the risk of keel damage if they do not have genetically-improved bone status; (c) feeding hens a particulate form of limestone from 15 weeks onwards can also increase bone strength and eggshell quality; (d) genetics, environment and nutrition all have independent and additive effects on bone status in laying hens but the relative effectiveness of these factors is genetics > environment > nutrition.

Incidence, pathology and prevention of keel bone deformities in the laying hen.

1. As a baseline study of the nature and incidence of keel deformities in laying hens, keel condition was examined in three different strains of hen from a total of 4 different caged environments (two commercial farms and two experimental farms). Incidence of keel deformity on farms in end of lay hens ranged from 2.6 to 16.7%. Only 0.8% of younger 15-week-old pullets had deformed keels. 2. Incidence of keel deformities was unchanged in 100 birds sampled from a free-range system compared to conventional caged siblings at the same farm. 3. Keel condition was also examined in 5 selected generations of a study involving the use of a body-weight-restricted selection index for skeletal improvement. Divergent selection for skeletal characteristics decreased incidence of keel deformity and improved radiographic density (RD) in high bone index (BI) hens compared to low BI hens in all selected generations. Male high BI keels were also improved compared to low BI. Shear strength measured in normal keels in generation 6 (G6) of the genetic study was improved in high BI hens compared to low BI hens. For all hens in the genetic study, those with normal keels had stronger tibiotarsus and humerus breaking strengths than hens with deformed keels. 4. Histopathology of keels representative of different deformities showed the presence of fracture callus material and new bone in all cases. This establishes that deformities are a result of trauma and are not developmental in origin. 5. Ash contents of keels, tibiae and humeri showed no differences between hens with normal and deformed keels. There were no differences in indicators of collagen cross-linkage in other bones between hens with normal keels and those with deformed keels. 6. It is concluded that lack of bone mass is the underlying cause of keel fracture and deformity in laying hens, rather than qualitative changes in bone, and that genetic selection can improve keel quality and prevent deformity.

High vitamin D3 requirements in broilers for bone quality and prevention of tibial dyschondroplasia and interactions with dietary calcium, available phosphorus and vitamin A.

1. Two experiments were carried out to investigate responses in performance and bone compositional and structural characteristics in broilers fed diets containing 4 concentrations of vitamin D3 (5, 20, 125 and 250 microg cholecalciferol/kg) at different concentrations of calcium, available phosphorus and vitamin A. 2. In experiment 1, body weight and tibia breaking strength were maximised at 14d with 250 microg vitamin D3/kg, tibia ash was maximised with 125 microg vitamin D3/kg. A high incidence of tibial dyschondroplasia (TD) was decreased to very low levels with 125 microg vitamin D/kg. 3. At 42d, performance and bone characteristics showed no response to vitamin D3 concentrations above 20 microg/kg. 4. Dietary vitamin A within the range 2-4 to 4.5 mg retinol/kg did not show any interaction with vitamin D3 status at either age. 5. In experiment 2, responses to vitamin D3 were strongly influenced by dietary calcium/available phosphorus. With 13 g calcium and 5 g available phosphorus/kg, performance and bone characteristics responded to vitamin D3 concentrations up to 125 microg/kg but more was needed at less optimal concentrations of calcium and available phosphorus. TD incidence was minimised with 250 microg/kg. 6. This study shows that high dietary concentrations of vitamin D3 can prevent TD. It is concluded that the vitamin D3 requirement of broilers up to 14 d of age at optimal dietary calcium and available phosphorus concentrations may be in the range 35 to 50 microg/kg for cortical bone quality and up to 250 microg/kg for prevention of TD. The vitamin D3 requirement for cortical bone quality after 14 d is not higher than 20 microg/kg. These requirements are much higher than earlier estimates and may be related to higher calcium requirements of modern broiler genotypes. Current regulations limiting maximum vitamin D3 concentrations in broiler starter diets may need to be reviewed.

Overview of bone biology in the egg-laying hen.

In young pullets, long bones elongate by endochondral growth. Growth plate chondrocytes proliferate, then hypertrophy, and are replaced by osteoblasts that form a network of trabecular bone. This bone is gradually resorbed by osteoclasts as the bone lengthens. Long bones widen, and flat bones are formed, by intramembranous ossification in which cortical bone formation by osteoblasts in the periosteal layer is accompanied by osteoclastic resorption at the inner endosteal surface. Growth of structural trabecular and cortical bone types continues up to the onset of sexual maturity in pullets. At this point, the large surge in estrogen changes the function of osteoblasts to forming medullary bone rather than structural bone. Medullary bone is a woven bone that acts as a labile source of calcium for eggshell formation. It lines structural bone and also occurs as spicules within the marrow cavity. It has little inherent strength but can contribute to fracture resistance. Osteoclasts resorb both medullary and structural bone so that during the period the hen remains in reproductive condition there is a progressive loss of structural bone throughout the skeleton, which is characteristic of osteoporosis. The increasing fragility of the bones makes them more susceptible to fractures. The dynamics of bone loss can be affected by a number of nutritional, environmental, and genetic factors. If the hen goes out of reproductive condition, estrogen levels fall, osteoblasts resume structural bone formation, and skeletal regeneration can take place.

Assessing bone mineral density in vivo: digitized fluoroscopy and ultrasound.

The genetic component of osteoporosis in caged laying hens is large, and a method for detecting hens susceptible to fracture could be useful in breeding programs. A radiographic absorptiometry film method was modified by video digitization from an image intensifier and computer analysis and termed digitized fluoroscopy (DF). Humeral and ulnar DF values were measured in 165 hens during lay. Relationships (P < 0.001) were seen between DF assessments from 25 wk onward and postmortem measurements at 70 wk. We conclude that DF can detect poor bones in hens early but is problematic. Quantitative ultrasound was also investigated. We measured amplitude-dependent speed-of-sound (Ad-SoS) in the third toe in hens. Nutritional studies revealed Ad-SoS values correlated with postmortem peripheral quantitative computerized tomography, (control group, r = 0.48, P < 0.001; treatment group, r = 0.39, P < 0.001). In caged and free-range hens, Ad-SoS correlated with shear strength (r = 0.33, P < 0.001, all hens) and radiographic density values (r = 0.53, P < 0.001, all hens) measured postmortem. The Ad-SoS values were higher in free-range than in caged hens (1,904 vs. 1,850 m/s, P < 0.001). Ad-SoS measurements were made in hens from a study where divergent genetic selection has produced high and low bone index lines with 92% difference in tibia strength. The value in high bone index hens was higher than in low bone index hens at 32 (P < 0.001), 42 (P < 0.001), 52 (P < 0.05), and 62 wk (P < 0.001) in generation 8. In an Ad-SoS heritability study, heritability estimates ranged from 0.15 to 0.39. We conclude that Ad-SoS is a heritable trait, reflects other bone measurements, and rapidly detects poor bone quality in hens.

Effects of dietary particulate limestone, vitamin K3 and fluoride and photostimulation on skeletal morphology and osteoporosis in laying hens.

1. Female chicks of a White Leghorn strain were fed three different diets from one day old: control, additional vitamin K3 (10 mg/kg), and a diet containing a combination of additional vitamin K3, sodium fluoride (10 mg/kg) and limestone in particulate rather than powdered form. At 16 weeks photoperiod was increased for half the birds from 8:16 L:D to 16:8 L:D immediately or by one hour per week to the same ultimate photoperiod for the other half. 2. Age at first egg was lower by 4.0 d for birds on the fast lighting regime but there were no overall effects of lighting on bone quality at either 25 or 70 weeks. 3. Additional vitamin K3 resulted in higher proximal tarsometatarsus cancellous bone volumes at 15 weeks and throughout the laying period compared with controls. Plasma osteocalcin concentrations were unaffected by vitamin K3 supplementation during growth. 4. The combination diet resulted in beneficial responses of 12 to 20% in most bone characteristics in hens at 70 weeks. The magnitude of these effects was similar to a previous study involving a particulate calcium source alone (Fleming et al., Poultry Science, 39: 434-440, 1998b). We conclude that the beneficial effects of the combined treatment over the lifetime of the hens were attributable mainly to the presence in the diet of a calcium source in particulate form.

Differences in composition of avian bone collagen following genetic selection for resistance to osteoporosis.

1. Collagen characteristics were compared in the tibiotarsus and humerus from 103 females and 38 males aged 68 to 72 weeks from the G6 generation of lines of laying hen selected for resistance or susceptibility to osteoporosis (high and low bone index (BI) lines). 2. Selection over the latest generation resulted in further divergence in the breaking strengths of humerus (from 12.3 to 21.8%) and tibia (from 22.3 to 37.3%) in hens. Males also showed line differences in bone strengths. 3. Plasma pyridinoline concentration was higher in hens in the low BI line, suggesting a greater rate of bone resorption in this line. 4. There were few differences between the lines in collagen and calcium concentrations in humerus and tibiotarsus cortical bone. 5. There were no differences between the lines in either sex in reduced immature collagen cross-link content of humerus or tibiotarsus. 6. Mature collagen cross-link content was higher in the high BI line in the male humerus but this effect was not apparent in the male tibiotarsus nor in either bone in the females. 7. Pyrrolic cross-link contents were higher in the high BI line in the female humerus and tibiotarsus and in the male tibiotarsus. 8. Over both lines combined, there were positive correlations between humeral and tibiotarsal pyrrole contents and strengths in females and between tibiotarsal pyrrole content and strength in males. 9. It is concluded that an increase in cross-linking, particularly pyrrolic cross-linking, in the collagen matrix contributes in part to the improvement in bone strength in the high BI line.

Prediction of breaking strength in osteoporotic avian bone using digitized fluoroscopy, a low cost radiographic technique.

Bone fragility in caged laying hens is a severe welfare problem, with fracture incidences in commercial flocks of up to 30% of all hens during their life. This fragility has been attributed to osteoporosis, the etiology of which is multifactorial in birds, as in humans, with genetic, environmental, and nutritional components. Greater understanding of the development of the disorder in hens could be obtained from the same kind of in vivo assessments available in human studies of osteoporosis. These high technology techniques for evaluation of bone mineral density (BMD), such as single or dual energy X-ray absorptiometry (SXA or DXA), quantitative computerized tomography (QCT), or attenuation by ultrasound (US), are so far not widely available to nonclinical researchers. We have modified an older X-ray film technique, radiographic absorptiometry (RA) by digitization of the analog video signal from a Philips BV-25 image intensifier, in single pulse fluoroscopy mode, and subsequent computer analysis with the public domain software package, NIH-Image 1.60. Compared with conventional RA, which uses standard X-ray film, our modified technique reduces X-ray exposure and allows the operator to digitize, store, and analyze many more images in a shorter time. We have called this modified technique "digitized fluoroscopy" (DF). In a longitudinal study of humeral radiographic density in a flock of 165 laying hens, significant relationships (P < 0.001) were observed between assessments made as early as 25 weeks, utilizing this DF technique in the humerus, and breaking strengths (and other postmortem indicators of osteoporosis) measured at 70 weeks. We conclude that DF can predict some eventual parameters of bone mass measured at 70 weeks from 25 to 40 weeks onward in bones from the same site in laying hens. The relationship between DF measurements made in the humerus and postmortem measurements of radiographic density and breaking strength made at another site (tibia) are less strong but still significant from 40 weeks onward.

Osteoporosis in cage layers.

Osteoporosis in laying hens is a condition that involves the progressive loss of structural bone during the laying period. This bone loss results in increased bone fragility and susceptibility to fracture, with fracture incidences of up to 30% over the laying period and depopulation not uncommon under commercial conditions. A major cause of osteoporosis is the switch in bone formation from structural to medullary bone at the onset of sexual maturity, but structural bone loss is accelerated by the relative inactivity of-caged birds. Allowing birds more exercise, as in aviary systems, results in better bone quality but may not decrease the overall fracture incidence. Good nutrition can help to minimize osteoporosis but is unable to prevent it. Best nutritional practice involves transferring birds to a higher calcium diet at lighting up rather than at first egg, providing a source of calcium in particulate form, and not withdrawing feed some days before depopulation. Breeding may be an effective way of combating ostoporosis. Some bone strength traits have been shown to be heritable, and divergent selection for resistance or susceptibility to osteoporosis has resulted in lines with markedly different bone characteristics. After three generations of selection, the lines differ by 19% for keel bone mineral density, 13% for humerus breaking strength, and 25% for tibia breaking strength and show a sixfold difference in fracture incidence under commercial breeding conditions. The difference in bone quality among the lines is maintained under different housing systems.

Expression patterns of chondrocyte genes cloned by differential display in tibial dyschondroplasia.

Tibial dyschondroplasia (TD) appears to involve a failure of the growth plate chondrocytes within growing long bones to differentiate fully to the hypertrophic stage, resulting in a mass of prehypertrophic chondrocytes which form the avascular TD lesion. Many biochemical and molecular markers of chondrocyte hypertrophy are absent from the lesion, or show reduced expression, but the cause of the disorder remains to be identified. As differentiation to the hypertrophic state is impaired in TD, we hypothesised that chondrocyte genes that are differentially expressed in the growth plate should show altered expression in TD. Using differential display, four genes, B-cadherin, EF2, HT7 and Ex-FABP were cloned from chondrocytes stimulated to differentiate to the hypertrophic stage in vitro, and their differential expression confirmed in vivo. Using semi-quantitative RT-PCR, the expression patterns of these genes were compared in chondrocytes from normal and TD growth plates. Surprisingly, none of these genes showed the pattern of expression that might be expected in TD lesion chondrocytes, and two of them, B-cadherin and Ex-FABP, were upregulated in the lesion. This indicates that the TD phenotype does not merely reflect the absence of hypertrophic marker genes, but may be influenced by more complex developmental mechanisms/defects than previously thought.

Nutrition: the integrative science.

1. Nutrition is a vitally important part of the production environment but also interacts with many other disciplines. 2. Modern breeding methods involving selection for a wider range of characteristics are likely to result in new genotypes with different nutritional needs. Geneticists and nutritionists will have to work closely together in the future to identify nutritional needs of these birds. 3. Collaboration between nutrition and other disciplines has been important in maintaining efficient poultry production and health and preventing or alleviating a number of metabolic disorders 4. There are many new challenges facing nutritionists to maintain health and performance in flocks under more exacting standards of bird welfare and human health. 5. A vigorous contribution from nutritional science and research will continue to be important for the future well-being of the poultry industry.

Inheritance of bone characteristics affecting osteoporosis in laying hens.

1. Heritabilities of a range of morphometric, radiological and strength characteristics were measured in the bones of end-of-lay hens. 2. Tibial strength (TSTR), humeral strength (HSTR) and keel radiographic density (KRD) were moderately to strongly inherited and were combined in a Bone Index which was used as a basis for selection. Data are available on 6 generations/cohorts of hens (n=1306), the last 3 of which are the progeny of divergently selected birds. 3. All bone characteristics used in the Bone Index responded rapidly to divergent selection and were strongly correlated with each other. In the last generation, the lines differed by 25% for TSTR, 13% for HSTR and 19% for KRD. The heritability of the index was 0.40. 4. There were no apparent genotype by environment interactions between birds housed at 2 different locations. 5. The incidence of bone fractures was significantly decreased in the line selected for high bone strength compared to the line selected for low bone strength. Humerus fracture incidence differed by a factor of 6 between the lines in the last generation. There was a strong quadratic relationship between tibia strength and overall fracture incidence (r2=0.92, P<0.01). 6. The results imply that selection for enhanced bone strength can be used as a long-term strategy for alleviating the problems of osteoporosis in laying hens.

A novel integral membrane protein is differentially expressed in the chick growth plate and maps to chromosome 1.

The growth plate is a specialised region of cartilage located at the growing ends of long bones in higher vertebrates. It is responsible for longitudinal bone growth and is under the control of many local and systemic factors. The growth plate consists of an orderly arrangement of small proliferative and larger mature hypertrophic chondrocytes. This paper describes the isolation by differential display of a 988-bp cDNA fragment derived from a transcript that is more highly expressed in proliferating rather than hypertrophic chondrocytes of the chick growth plate. Using 3' RACE, a further 939 bp of cDNA sequence was obtained. The 1.9 kb sequence contains a 924-bp open reading frame encoding an unknown 308 amino acid protein. This protein has a putative transmembrane domain near its N-terminus and three dileucine motifs at its carboxy tail. This gene was expressed in all other tissues examined. A polymorphism was identified by SSCP analysis and the gene was mapped to the centromeric region of the short arm of chicken chromosome 1, close to the locus for autosomal dwarfism.

Optimal dietary concentration of vitamin E for alleviating the effect of heat stress on egg production in laying hens.

1. The effects of different dietary concentrations of vitamin E (alpha-tocopherol acetate) were investigated on laying hens exposed to chronic heat stress at 32 degrees C from 26 to 30 weeks of age. 2. Diets containing 5 dietary concentrations of vitamin E (a control diet containing 10 mg alpha-tocopherol/kg or this diet supplemented to contain 125, 250, 375 and 500 mg alpha-tocopherol/kg) were fed to 335 birds. Half of the birds received the supplemented diets for only 4 weeks before the heat stress period (short supplementation duration, SSD) and were fed on the control diet for a further 12 weeks. The remaining birds were fed on the supplemented diets throughout the experiment, 4 weeks before, 4 weeks during and 8 weeks after the heat stress period (long supplementation duration, LSD). 3. Egg production was significantly higher during (80-6 vs 68.9%, P<0.02) and after (75.3 vs 62.7%, P<0.02) the period of stress in the LSD group fed on the diet containing 250 mg vitamin E/kg compared with the group fed on the control diet. LSD birds given 375 and 500 mg vitamin E/kg also had higher egg production than control birds during heat stress but the differences failed to reach significance (74.6 vs 68.9% and 77.1 vs 68.9% respectively). In the SSD groups, mean egg production of the birds given the diets supplemented with 125 mg vitamin E/kg or more was significantly different from the control group after heat stress (70.3 vs 62.7%, P<0.05). Egg weight and food intake were similar in all the groups. 4. Plasma and liver vitamin E concentrations were proportional to the vitamin E intake before the stress period, dropped during heat stress in the SSD groups but were maintained at concentrations closer to those observed before heat stress in the LSD groups. 5. It is concluded that a dietary supplement of 250 mg vitamin E/kg provided before, during and after heat stress is optimum for alleviating, at least in part, the adverse effects of chronic heat stress in laying hens.

Ascorbic acid-induced chondrocyte terminal differentiation: the role of the extracellular matrix and 1,25-dihydroxyvitamin D.

Chondrocyte terminal differentiation is associated with cellular hypertrophy increased activity of plasma membrane alkaline phosphatase and the synthesis of collagen type X. The hypertrophic phenotype of cultured chondrocytes can be stimulated by ascorbic acid but the underlying mechanisms for this phenotypic change are unclear. As ascorbic acid is central to many hydroxylation reactions, the possibility was examined that its pro-differentiating effects are mediated by its effects on collagen and vitamin D metabolite formation. In vitro studies indicated that ascorbic acid-induced chondrocyte alkaline phosphatase activity was inhibited by the addition of both collagen and proteoglycan synthesis inhibitors. The addition of arginine-glycine-aspartic acid (RGD)-containing peptides also resulted in lower alkaline phosphatase activity. Chicks supplemented with dietary ascorbic acid had higher concentrations of both collagen and proteoglycans within their growth plates but the chondrocyte maturation rate was unaltered. No evidence was obtained to suggest that ascorbic acid-induced collagen production was mediated by lipid peroxidation. In addition, supplementation with dietary ascorbic acid resulted in higher serum 1,25-dihydroxyvitamin D3 concentrations and increased chondrocyte vitamin D receptor number. Ascorbic acid-treated chondrocytes maintained in vitro also had increased vitamin D receptor numbers but chondrocyte receptor affinity for 1,25-dihydroxyvitamin D3 was unaltered. These results indicate that ascorbic acid promotes both chondrocyte matrix production and 1,25-dihydroxyvitamin D3 synthesis, accompanied by upregulation of the vitamin D receptor. Thus, ascorbic acid may be causing amplification of the vitamin D receptor-dependent genomic response to 1,25-dihydroxyvitamin D, resulting in promotion of terminal differentiation. Strong evidence is provided to support the hypothesis that ascorbic acid-induced chondrocyte terminal differentiation is mediated by interactions between integrins and RGD-containing cartilage matrix proteins.

Bone structure and strength at different ages in laying hens and effects of dietary particulate limestone, vitamin K and ascorbic acid.

1. A range of bone structural and strength characteristics was determined in laying hens at 15, 25, 50 and 70 weeks of age. The birds were fed up to 25 weeks on diets supplemented with additional vitamin K (10 mg menadione/kg) or ascorbic acid (250 mg/kg) or up to 70 weeks on diets containing limestone in powder or particulate form. 2. There were important effects of age on all bone characteristics. Between 15 and 25 weeks there was a rapid loss of cancellous bone and a rapid accumulation of medullary bone in the proximal tarsometatarsus (PTM). These changes continued at a slower rate up to 70 weeks. Cancellous bone content of the free thoracic vertebra (FTV) also declined after 15 weeks. 3. Breaking strengths of tibia and humerus did not change between 15 and 25 weeks but decreased later in lay. 4. None of the nutritional treatments affected bone characteristics at 15 weeks of age. 5. Increasing the dietary vitamin K supplement from 2 to 12 mg menadione/kg increased cancellous bone volume in the PTM at 25 weeks. 6. Dietary ascorbic acid did not affect any of the bone characteristics measured up to 25 weeks. 7. Particulate limestone resulted in a smaller loss of cancellous bone between 15 and 25 weeks and increased accumulation of medullary bone in the PTM. Breaking strength of the tibia and radiographic densities of tibia and keel were also improved. 8. It is concluded that patterns of bone loss over the lifetime of laying hens vary, depending upon the bone type. Feeding a particulate source of calcium can help to alleviate some of the characteristics of osteoporosis. Supplementation with extra vitamin K may also be beneficial.