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.     

Bone structure and breaking strength in laying hens housed in different husbandry systems

1. Bone structure and breaking strength were measured in hens that had been housed throughout a laying year in battery cages or in Perchery, Naturel or Litter and Wire husbandry systems.

2. Battery caged hens had the poorest bones, as assessed by measurements of cancellous bone volume, radiographic density, cortical thickness and three‐point breaking strength.

3. Humeri from birds in the Litter and Wire system were less dense radiographically and weaker than those from Perchery or Naturel birds but leg bone characteristics were similar with these three systems.

4. There were no differences in bone characteristics between birds in Perchery and Naturel systems.

5. There were strong correlations between radiographic densities and strengths of contralateral humeri and tibiae over all husbandry systems. Humerus structural and strength characteristics may be the best criteria of osteoporosis in hens.

6. It is concluded that the extent of movement allowed by different husbandry systems affects structural bone loss and bone strength in laying hens.

7. It is further concluded that the breaking strength of a hen's bone is closely related to morphometric measures and radiographic density of its structural components.     

Limb bone strength and movement in laying hens from different housing systems

Walking and bouts of wing movement performed by 61-week-old ISA brown laying hens which had been taken from one batch of eggs and then kept in three different housing systems which allowed a gradation in spatial freedom, were recorded. After slaughter the breaking strengths of the humerus and tibia of birds from each system were measured. Birds from battery cages exhibited the fewest limb movements and had the weakest bones, their humeri having only 54 per cent of the strength of those of birds from a perchery. Birds from the Elson terrace system were heavier and had a stronger tibia than cage birds. Compared with perchery birds, terrace birds had weaker humeri and also performed fewer wing movements. The results indicate that the amount of movement possible for laying hens in battery cages was insufficient to avoid levels of osteopenia and consequent bone fragility much greater than in birds kept in the perchery and Elson terrace systems.     

Keeping laying hens in furnished cages and an aviary housing system enhances their bone stability

1. Tibia and humerus breaking strength of Lohmann Silver hybrids kept in conventional cages, furnished cages and an aviary with outdoor run were examined in two production cycles. Each trial lasted a full laying period; feeding, management and healthcare were identical for all hens. In both trials bone strength was investigated at the end of laying months 6, 9 and 14. 2. The objective was to determine if bone strength increases when hens are kept in alternative housing systems, especially in furnished cages, and whether hen age affects bone stability. 3. The results indicated that housing system influenced bone breaking strength, which was consistently higher for hens in the aviary compared to hens in conventional and furnished cages. Furthermore, humerus breaking strength was higher for hens in furnished cages compared to conventional cages. No significant difference regarding tibia breaking strength was found between conventional and furnished cages. 4. Our results showed that lack of exercise contributed to the problem of weak bones more than did calcium depletion from eggshell formation. 5. Tibia breaking strength increased during the last third of the production cycle, whereas humerus breaking strength remained unaffected by hen age. 6. Genetic group affected only tibial bone breaking strength, which was lower overall in genetic group A than in group B, which in turn was lower than group C. 7. The increased bone strength in the aviary and in the furnished cages probably reduced the incidence of recently broken bones in these systems compared to the conventional cages. This increase in bone strength can be regarded as an improvement in welfare. Furnished cages, like the aviary system, might be considered an alternative housing system for laying hens, because both resulted in enhanced bone strength.     

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.     

Bone histomorphometry in 1,25(OH)2D3- and vitamin D3-treated aged laying hens.

Aged laying hens at oviposition were given four levels of dietary vitamin D3 and three levels of 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) to determine the effects of the additives on the medullary and endosteal bone. Oxytetracycline hydrochloride labeling and histomorphometry of undecalcified sections were used in the determination of changes in medullary bone and endosteal surface of mid-diaphyseal tibiotarsal bones. Treatment with 1,25(OH)2D3 increased mineralized medullary bone area, endosteal surface length, amount of tetracycline labeling in medullary and endosteal bone, total number of osteoclasts on endosteal and medullary bone, and appositional rate of mineralizing bone. Vitamin D3 increased mineralized medullary bone area and total tetracycline labeling in endosteal bone but decreased endosteal mineral apposition rate. Medullary bone matrix with no mineral was observed only in the group receiving neither supplement. These areas were much larger than typical osteoid seams usually seen in trabecular bone. Both 1,25(OH)2D3 and vitamin D3 were effective in stimulating bone formation and retention of mineral within medullary bone matrix at oviposition.     

Radiodensity in the central cavity of humeri in high-producing non-commercial laying hens

1. Line differences in the incidence of the presence of radiodense material in the central cavities of humeri of 6 different lines of laying hens were observed using radiographs. 2. Internal humeral radiodense material was observed in over 60% of hens in two lines and 18% of hens in two other lines measured at 65 weeks of age. There was no difference in the incidence (15, 16%) between the last two lines observed at 47 weeks of age. 3. In the 4 lines studied at 65 weeks of age the presence of radiodense material was associated with increased humerus dry weight and ash weight in two lines, while percentage-ash was reduced in all 4 lines. 4. Results from this and previous studies suggest that humeral medullary bone could be an appreciable source of variability in experimental data collected to assess skeletal health.     

The effect of intra-articular methylprednisolone acetate and exercise on equine carpal subchondral and cancellous bone microhardness

Dorsal carpal osteochondral injury is a major cause of lameness in horses undergoing high intensity training. Intra-articular corticosteroid treatment is used commonly to manage exercise-associated articular pain, but its use remains highly controversial in the equine athlete. This project, therefore, aimed to compare the mechanical properties of intra-articular MPA and diluent-treated middle carpal subchondral and cancellous bone in horses undergoing a short-term treadmill exercise programme. It was hypothesised that subchondral and cancellous bone mechanical properties are influenced by intra-articular administration of methylprednisolone acetate (MPA). Eight 2-year-old female horses had MPA or diluent administered into contralateral middle carpal joints at 14 day intervals, for a total of 4 treatments per horse. Horses underwent a standard treadmill exercise protocol until euthanasia (Day 70). Standard sites were located on the dorsal aspect of third, radial and intermediate carpal bones. Osteochondral samples from each test site were divided into subchondral bone and cancellous bone portions. These were dried, resin-embedded and gold-coated. Microhardness measurements were obtained at each test site. No significant effect of intra-articular treatment was detected. At each site, cancellous bone trabecular struts had an 18-19% higher microhardness value than the overlying subchondral bone. These findings indicate that intra-articular administration of MPA at this dose has no effect on subchondral or cancellous bone adaptation to short-term exercise and, therefore, on the propensity of carpal bones to injury. Further investigation into the calcified cartilage layer, effect of different corticosteroid preparations and diffusion of medication are required.     

Ipriflavone reverses the adverse effects of a low-calcium diet on the histology of the tibia in caged layers

1. Ipriflavone (IP) has been widely studied in humans and is effective for inhibiting osteoclastic bone resorption and enhancing osteoblastic bone formation. The aim of this study was to investigate the effects of IP on the structural histology and histomorphometric parameters of tibia in laying hens fed on a low-calcium diet.

2. A total of 200 Hy-Line Brown laying hens, 24 weeks of age, were divided randomly into 4 groups which were fed on one of 4 diets (CaN, standard diet; CaL, low-calcium diet; IP1, low-calcium diet + 8 mg/kg of IP; IP2, low-calcium diet + 20 mg/kg of IP) for 60 d after which 10 hens of each group, chosen at random, were killed and the tibiae were processed and assessed under a fluorescent microscope

3. The cortical bone showing resorption cavities had become porous, and medullary bone did not completely fill the marrow cavity in the CaL group compared with the CaN group. However, in the IP2 group, the cortical bone showed a structure comparable to the CaN group with an absence of resorption cavities, and more medullary bone filled the marrow cavity compared with the CaL group.

4. The CaL hens had significantly lower trabecular bone area, trabecular perimeter, per cent trabecular area, trabecular number and trabecular thickness and significantly wider trabecular separation compared with CaN. These results indicated that the low-calcium diet led to increased loss of tibial bone, eventually causing osteoporosis. However, compared with the CaL group, the IP2 group caused a significant increase in trabecular bone area and bone quality.

5. It is concluded that even in hens fed on a low-calcium diet, the addition of 20 mg/kg IP exerted beneficial effects on histological structure and structural histomorphometric parameters of tibia, indicating an alleviation of caged layer osteoporosis (CLO).     

Expression and localization of estrogen receptors α and β mRNA in medullary bone of laying hens

The aim of this study was to observe the expression and localization of estrogen receptor (ER) α and ER β mRNA in the medullary bone of laying hens. First, medullary bone, liver, kidney, and shell gland of the oviduct tissues were dissected from laying hens. Then, the total cellular RNA was isolated from each tissue specimen, and the ER α and ER β mRNA expression was observed using semiquantitative RT‐PCR. Second, the localization of ER α mRNA in the medullary bone was detected with in situ hybridization using digoxigenin‐11‐UTP‐labeled cRNA probes. As a result, the expression of ER α mRNA was higher than that of ER β mRNA in the medullary bone, liver, and shell gland of the oviduct from laying hens. In the kidney, ER α mRNA expression was lower than that of ER β mRNA. The expression pattern of ER α and ER β mRNA of the medullary bone was similar to that of the shell gland of the oviduct. Moreover, ER α mRNA was intensively expressed in osteoblasts on the medullary bone surface and bone marrow stromal cells but was not expressed in osteoclasts. These results suggest that in medullary bone, estrogen action may be regulated not by ER β but by ER α.     

Computed tomographic and radiographic morphology of the pastern and coffin joints of One-Humped Camel (Camelus dromedarius)

The objective of the current study was to describe the structures of the pastern and coffin joints in dromedary camel using x-ray, bone and soft tissue windows computed tomography (CT) and three-dimensional volume rendering (3DVR) of CT imaging. 3DVR of CT was obtained at the slight flexed dorsal view, plantar view, dorsolateral view and lateral view which explained all the surfaces and structures of the digit bony parts even the parts of the articular surface. The processed images of 3DVR of CT showed different patterns of the cortical, cancellous, subchondral bones and medullary cavity of the bones of the digits. The present study showed clearly all the hard and soft tissues in the pastern and coffin joints of the camel in CT images; however, the plantar ligaments of the pastern joint and ligaments of the navicular cartilage identified on CT images. The CT soft tissue window visualized the joint cavity and their pouches and tendon sheath of the flexor tendons better than the bone window CT. The radiographic, CT and 3D images could be used as a normal reference for the interpretation of some clinical diseases in the pastern and coffin joints of the camel.     

Axial pull-out strength of 3.5 cortical and 4.0 cancellous bone screws placed in canine proximal tibias using manual and power tapping

An in vitro experimental cadaveric mechanical testing study was performed using 20 radiographically mature dogs, weighing between 18-33 kg. The aim of the study was to compare the axial pull-out strength of 3.5 mm cortical and 4.0 mm cancellous bone screws inserted in the canine proximal tibia using manual and power tapping techniques. 3.5 cortical and 4.0 cancellous bone screws were inserted in canine cadaver proximal tibiae using a manual or power tapping technique. The screws were extracted using a servohydraulic materials testing machine in order to measure axial pull-out strength. Axial pull-out strength was recorded relative to the total bone width and total cortical width of each tibia. The mean axial pull-out strength for all constructs was 717.8+/-56.5 N without any statistically significant difference among groups (p=0.4183). The groups were equal in animal body weight, cortical width and total bone width (p=0.2808). The axial pull-out strength in proportion to cortical and total bone width was not significantly different among groups (p=0.5318). Axial pull-out strengths of 3.5 mm cortical and 4.0 mm cancellous bone screws inserted in the proximal tibial metaphysis were not significantly different. Axial pull-out strength was not affected by the use of power tapping in either screw type.     

Histochemical identification of oestrogen target cells in the medullary bone of laying hens

1. Oestrogen target cells in the medullary bone of laying hens were examined using a histochemical method with fluorescein isothiocyanate labelled oestradiol. 2. Specific fluorescence was found to occur strongly in osteoblasts and weakly in osteoclasts on the medullary bone surface. 3. This study suggests that osteoblasts on the medullary bone surface are oestrogen target cells.     

Multifocal idiopathic pyogranulomatous bone disease in a dog

Disseminated, multifocal idiopathic pyogranulomatous bone disease in a five-year-old neutered female crossbred cattle dog is described. The dog was presented for neck pain and reluctance to exercise. Radiography demonstrated polyostotic bone disease, the predominant change consisting of a mottled pattern of osteosclerosis interspersed with punctate foci of osteolysis. Numerous bones of the axial and appendicular skeleton were affected including cervical vertebrae, ribs, scapulae and humeri. Biopsy revealed sterile pyogranulomatous osteomyelitis. This was confirmed at necropsy which showed that lesions were restricted to bone. Idiopathic granulomatous or pyogranulomatous conditions have been described in the dog in relation to skin but not bone. In people, idiopathic granulomatous bone disease is one of a group of conditions collectively called histiocytosis X, which are thought to be immunoregulatory disorders related to Tlymphocyte suppressor cell deficiency and Langerhans' cell proliferation.     

In Vitro Pullout Strength of Screws Inserted in Adult Equine Third Metacarpal Bone After Overdrilling a 4.5-mm Threaded Insertion Hole

Objective—To determine and compare the in vitro pullout strength of 5.5-mm cortical versus 6.5-mm cancellous bone screws inserted in the diaphysis and metaphysis of adult equine third metacarpal (MCIII) bones, in threaded 4.5-mm cortical bone screw insertion holes that were then overdrilled with a 4.5-mm drill bit to provide information relevant to the selection of a replacement screw if a 4.5-mm cortical screw is stripped. Study Design—In vitro pullout tests of 5.5-mm cortical and 6.5-mm cancellous screws in equine MCIII bones. Sample Population—Two independent cadaver studies each consisting of 14 adult equine MCIII bones. Methods—Two 4.5-mm cortical screws were placed either in the middiaphysis (study 1) or distal metaphysis (study 2) of MCIII bones. The holes were then overdrilled with a 4.5-mm drill bit and had either a 5.5-mm cortical or a 6.5-mm cancellous screw inserted; screw pullout tests were performed at a rate of 0.04 mm/second until screw or bone failure occurred. Results—In diaphyseal bone, the screws failed in all tests. Tensile breaking strength for 5.5-mm cortical screws (997.5 ± 49.3 kg) and 6.5-mm cancellous screws (931.6 ± 19.5 kg) was not significantly different. In metaphyseal bone, the bone failed in all tests. The holding power for 6.5-mm cancellous screws (39.1 ± 4.9 kg/mm) was significantly greater than 5.5-mm cortical screws (23.5 ± 3.5 kg/mm) in the metaphysis. There was no difference in the tensile breaking strength of screws in the diaphysis between proximal and distal screw holes; however, the holding power was significantly greater in the distal, compared with the proximal, metaphyseal holes. Conclusions—Although tensile breaking strength was not different between 5.5-mm cortical and 6.5-mm cancellous screws in middiaphyseal cortical bone, holding power of 6.5-mm cancellous screws was greater than 5.5-mm cortical screws in metaphyseal bone of adult horses. Clinical Relevance—If a 4.5-mm cortical bone screw strips in MCIII diaphyseal bone of adult horses, either a 5.5-mm cortical or 6.5-mm cancellous screw, however, would have equivalent pullout strengths. A 6.5-mm cancellous screw, however, would provide greater holding power than a 5.5-mm cortical screw in metaphyseal bone.     

Comparison of bone healing,as assessed by computed tomography,following tibial tuberosity advancement in dogs with and without autogenous cancellous bone grafts

Abstract

AIMS: To objectively compare measures of bone healing, using computed tomography (CT) in dogs following bilateral tibial tuberosity advancement (TTA), between tibiae treated with and without autogenous cancellous bone grafts.

METHODS: Ten dogs with bilateral cranial cruciate ligament disease requiring surgical stabilisation were prospectively recruited to undergo single-session bilateral TTA, with only one, randomly assigned, tibia receiving bone graft in the osteotomy deficit. Bone healing at the osteotomy site was assessed using CT performed 38–70 days post-operatively. CT images were evaluated using both objective measurements of osseous bridging and subjective evaluation by six radiologists. Repeated measures ANOVA was used to compare the objective outcomes between the grafted and non-grafted tibiae.

RESULTS: The mean percentage of the osteotomy deficit bridged at the lateral cortex was greater in grafted (77.6, SD 35.2%) compared to non-grafted (63.0, SD 36.5%) tibiae (p=0.001), but did not differ at the medial cortex (p=0.1). The mean minimum callus width was greater in grafted (7.2, SD 3.3 mm) compared to non-grafted (3.6, SD 2.9 mm) tibiae (p<0.001). There was no difference in mean attenuation (measured in Hounsfield units) of the callus between grafted and non-grafted tibiae (p=0.5). The grafted tibia was deemed to have superior bone healing in 50/60 subjective assessments made by radiologists.

CONCLUSIONS: Superior osseous bridging was detected by CT analysis following TTA using autogenous cancellous bone grafts compared with no graft. This was shown by greater bridging percentage at the lateral cortex and formation of a broader callus. Qualitative assessments made by six radiologists also supported the conclusion that bone healing was improved by use of autogenous cancellous bone graft. CT was a useful method for assessing evidence of bone healing following TTA.

CLINICAL RELEVANCE: These findings justify the application of autogenous cancellous bone graft to augment healing following TTA in dogs.     

Guided bone regeneration with beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone membrane in partial defects of canine humerus

This study was performed to evaluate the effect of beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone (TCP/PLGC) membrane in the repair of partial bone defects in canine proximal humerus. Three adult mixed-breed dogs were used during the experimental period. The length of the defect was quarter of the full length of humerus, and width of the defect was quarter of middle diameter of the lateral aspect of humerus. The humeri of each dog were divided into treatment (TCP/PLGC) and control groups. The defect was covered with TCP/PLGC membrane in treatment group. To evaluate regeneration of the bone, computerized tomography (CT) and histopathologic examination were performed. The radiopaque lines were appeared at the original defect sites in TCP/PLGC group but below the original site in control at 4th week. Radiopacity and thickness of the defect sites, and radiopaque lines were more increased at 8th week than those of 4th week. Histopathologic findings revealed fibrous connective tissue migration into the defect and the migration inhibited the structure of new cortex to be placed in the original level in control whereas new cortex growth was found in the level of original line in TCP/PLGC group. However, the new cortical bone in the TCP/PLGC group was thinner and less organized than the adjacent intact cortex, and the amount of new cancellous bones were also scanty. The result suggested that TCP/PLGC membrane is a good guided bone regeneration material to restore the original morphology of humerus in partial defect.     

Combination of a Corticai Allograft With a Cancellous Autograft in the Canine Tibia

This study was undertaken to determine what difference, if any, occurred when autogenous cancellous bone was placed in the medullary canal of a fresh, 4-cm cortical allograft compared to a fresh cortical allograft alone. Twelve mature dogs were used. A tubular cortical graft was placed in the midshaft of both tibias. The right tibia received the combination graft while the left tibia received a fresh cortical allograft only. Study periods ranged from one week to 17 weeks. The progress of the grafts was assessed by technetium-99m radionuclide bone scans, radiographs, gross and histological examination, and tetracycline labeling. Results indicate that autogenous cancellous bone placed in the medullary canal of a fresh cortical allograft was beneficial to the ingrowth of new blood vessels and incorporation of the cortical graft.     

Morphology of medullary bone during the egg formation cycle

The medullary bone of birds is known to function as a calcium store for egg shell production, but it is not known if the stage of shell formation within the egg formation cycle is reflected by the thickness of osteoid seams on the medullary bone. Adult laying hens were killed at two points in the formation cycle to determine the effects of shell formation on medullary bone using histological techniques. No differences in medullary bone volume or osteoid thickness were apparent at either of the two points. Fluorochrome bone labelling techniques demonstrated, however, that less medullary bone mineralisation occurred during the active period of shell formation.     

Morphometric Study of the Equine Navicular Bone: Age-Related Changes and Influence of Exercise

Navicular bones from the four limbs of 95 horses, classified in 9 categories, were studied. The effects of age on navicular bone morphometry and histomorphometry were estimated, after adjustment of the data to even out the effects of front and rear limbs, morphometrical type, sex, weight, and size.All the external measurements of the navicular bone decreased significantly with increasing age. From the histomorphometrical data, cortical bone volume decreased with age in most horses, whereas cancellous bone volume and, in particular, the marrow spaces increased. The increase in the cancellous bone volume could have resulted from tunnelling of the internal part of the cortex, which converted it progressively into a porous trabecular-like structure. Trabecular bone volume also decreased with age and the trabecular lattice changed dramatically to become disconnected in aged horses. These observations corresponded closely to those reported for ageing of the skeletal system in humans.However, in sporting horses, the navicular cortical bone volume increased with age and the cancellous bone volume decreased. Exercise appeared to have decreased bone resorption and increased bone formation at the endocortical junction. The cancellous bone architecture was also improved, in that the trabecular lattice and trabecular bone volume remained unchanged in aged sporting horses. Our findings confirmed that exercise may be good practice to prevent age-related bone loss.