Microtomographic Parameters and Nanoindentation of the Hoof of Girolando Cattle

The aim of this study was to describe the microstructure of the pigmented and depigmented hoof capsule of Girolando cattle by bi‐ and tridimensional microtomography and nanoindentation, analysing the possible relation between these findings and the susceptibility of such animals to podal diseases. To carry out the microtomography and the nanoindentation, duplicated samples were collected from the dorsal wall, abaxial wall and pre‐bulbar sole of the hoof capsule. Material collection was performed in 40 medial digits of thoracic limbs and 40 lateral digits of pelvic limbs. The bidimensional microtomography showed that the dorsal wall of the thoracic and pelvic limbs presented higher density, followed by the abaxial wall, and finally by the sole, with the lowest density. Moreover, the hoof capsule of cows of Girolando breed is a compact, non‐porous material, and constituted by extratubular and intratubular keratin. By the tridimensional microtomography, it was possible to measure the angles of the corneal tubules in relation to the periople and the claws in the different regions of the hoof capsule, which were 90° for the dorsal wall, 55° for the abaxial wall and 70° for the sole. The tridimensional microtomography also showed corneal tubules of different diameters: 17, 51, 85, 119 and 153 μm. The nanoindentation test, when performed in different regions of the hoof capsule, did not reveal significant difference of Vickers hardness in the evaluated areas. However, we verified a larger elastic module of these regions on the transversal cut of the corneal tubules compared to the longitudinal cut.     

Mineral composition and microstructure of the abaxial hoof wall in dairy heifers after biotin supplementation

The aim of this study was to evaluate the effect of daily biotin supplementation on the mineral composition and microstructure of the abaxial hoof wall in dairy heifers. The heifers were housed on a concrete floor and fed for weight gain more than 800 g per day, which is a challenging environment for the hoof. Twelve crossbred dairy heifers (Jersey × Holstein) were divided into two treatment groups. Animals in the control group (n = 6) received a diet without supplemental biotin, while the heifers in the biotin-supplemented feed group (n = 6) each received 20 mg of biotin daily for 120 days. Samples of the abaxial hoof wall were collected from the outer claw of the fore and hind limb, before and after supplementation. The samples were evaluated by X-ray fluorescence spectrometry, computed microtomography, atomic force microscopy and confocal laser scanning microscopy. Biotin supplementation increased the sulphur content and decreased the calcium and potassium content in the abaxial hoof wall. Biotin treatment also increased the percentage of horn tubules with smaller diameter marrow (17–51 µm). However, biotin did not influence the surface relief of the hoof wall, suggesting that its action is limited to the inner layers of the stratum corneum. Daily supplementation with 20 mg of biotin promoted changes in the mineral composition and microstructure of abaxial hoof wall of crossbred dairy heifers. These findings suggest biotin supplementation improves hoof quality and may help to understand the function of biotin in the stratum corneum.     

External characteristics of the lateral aspect of the hoof differ between non-lame and lame horses

During a pre-purchase examination (PPE) there is always a debate about how clinical findings of the hoof different from ideal should be interpreted in relation to future lameness risk and/or unsuitability of the horse for the potential purchaser. The objectives of this study were to describe and compare external angular measurements, linear ratios and hoof capsule characteristics of non-lame and lame feet. Photographs of feet from 300 horses with foot pain and 25 non-lame horses were analysed. Hoof wall, heel and coronary band angles and hoof wall length and height, weight-bearing length, coronary band length and height of the coronary band at dorsal and palmar locations were measured and expressed as linear ratios.Mean hoof wall, heel and coronary band angles were larger in lame compared with non-lame feet; only the ratio of dorsal to palmar coronary band heights and the shape of the coronary band were significantly different between lame and non-lame horses. Growth rings were divergent and horn tubules were non-parallel in lameness of >3 months. At a PPE, a larger ratio of dorsal to palmar coronary band heights in one limb may be indicative of previous lameness in that foot. Changes in coronary band shape and divergent growth rings and horn tubules would suggest a longer duration.     

The hardness of horn in different segments of the bovine claw

Hardness of bovine hoof horn was tested as ball indentation hardness and as shore D hardness post mortem in different segments of the hoof wall, in the sole and the hard bulb of sound claws of 10 Austrian Holstein Friesian cows. Both methods of hardness determination showed corresponding results, with shore D hardness between 52.2 and 63.9 hardness units (hu) and ball indentation hardness between 11.2 N/mm2 and 24.3 N/mm2. Bovine hoof horn becomes significantly softer from the coronary band towards the weight bearing border (vertical decrease) and from the dorsal wall towards the heel (horizontal decrease). Decreasing hardness was associated with decreasing dry matter content. Measurements of the claw capsule showed thickness of the hoof wall increasing from the coronary border towards the sole. In dorsopalmar/-plantar direction, bovine hoof wall at the weight bearing border decreases towards the heel.     

Modulus of elasticity and dry-matter content of bovine claw horn affected by the changes of chronic laminitis

The mechanical properties of horn samples from 22 hind claws with chronic laminitis were determined in adult Austrian Fleckvieh cows. The resistance to deformation was quantified as the modulus of elasticity (E). Tension tests revealed mean E values of 520MPa for the dorsal wall, 243MPa for the abaxial wall, 339MPa for the axial wall and 97MPa for the sole. E tended to be lower in laminitic horn than in sound horn in all segments tested, with the difference being largest in the abaxial wall. The mean dry-matter content (DMC) of the laminitic claws was 75.8% in the dorsal wall, 75.86% in the abaxial wall, 71.15% in the axial wall and 69.28% in the sole. These values are generally comparable to those for sound claws except in the axial wall. Further, E and DMC were only correlated in the axial wall. Chronic laminitis leads to a low resistance of claw horn to mechanical insults in the dorsal wall, abaxial wall and sole, and to the loss of a correlation between the E and DMC in these segments. The reason for these alterations is therefore not increased ingress of moisture, but must be due to changes in the microstructure, biochemical components and/or horn formation by the diseased dermis.     

Hoof growth and wear in pasture-fed dairy cattle

Hoof growth and wear studies were conducted on ten spring-calving cows over 12 months. The rates of hoof wall growth and wear were lower in autumn and winter than during spring and summer. Both were greater in 2-year-old than in mature cows (p<0.05). Sole wear occurred most rapidly along the abaxial edge of the weight-bearing surface, less quickly in the toe and heel areas, and most slowly in the mid-sole region. Both hoof wall wear and sole wear were greater in lateral digits than in medial digits (p<0.01). Sole concavity, measured using a profile gauge, was greater in 2-year-olds prior to calving than in mature-aged cows (p<0.05) but this difference disappeared soon after calving as concavity was lost in the younger cows. Medial digits almost always had greater concavity than lateral digits. Sole concavity measurements were also made on the hind digits of 24 autumn-calving cows on four occasions at 2-monthly intervals. Prior to calving, concavity was much greater in these cows than in the spring-calving cows. However, concavity was lost rapidly following calving and concavity measurements were soon similar to those found in the spring-calving cows.     

Fenestration of the abaxial hoof wall and implantation of gentamicin-impregnated collagen sponges for the treatment of septic arthritis of the distal interphalangeal joint in cattle.

A novel surgical approach to the distal interphalangeal joint of cattle, through the abaxial hoof wall, for the treatment of septic arthritis is described. In seven cattle a rectangular piece of horn (15 x 40 mm) was excised from the abaxial hoof wall and the joint was lavaged with Ringer's solution. Infected and/or necrotic tissue was removed from the joint and gentamicin-impregnated collagen sponges were introduced. The piece of horn was reattached, a rubber block was glued to the contralateral claw, and a bandage applied. Aftercare consisted of changes of bandage, systemic administration of antimicrobial drugs and box rest. At follow-up, six of the cattle were sound but one cow which had been slaughtered five months after surgery had remained slightly lame.     

Ultrasonographic Imaging and Thickness Measurement of the Sole Horn and the Underlying Soft Tissue Layer in Bovine Claws

Ultrasonographic imaging and thickness measurements of sole horn and underlying soft tissue layer were evaluated in 100 bovine claw specimens. The claws were trimmed and examined in longitudinal planes using a 7.5 MHz linear transducer. The sole horn, underlying soft tissue layer and distal surface of the distal phalanx of all claws were clearly visualized. The sole horn had a heterogeneous hypoechoic appearance; the underlying soft tissue layer was predominantly anechoic. Corium and subcutis could be differentiated. The best imaging of these structures was achieved in claws with less than 10 mm sole horn thickness and soft sole horn. Imaging of the dorsal wall structures was more difficult. In each claw, B- and A-mode measurements of horn thickness of sole and dorsal wall and distance from outer claw surface to distal phalanx surface were made at three marked points on the weightbearing surface and at one point on the dorsal wall. In 10 claws, CT-images were taken and the horn density was determined. All claws were then frozen, transected and anatomical reference measurements were made after thawing. Statistical correlation coefficients of 0.88 and higher were found for B-mode ultrasonographic and anatomical measurements of sole horn thickness. A-mode proved not to be useful for the sole horn thickness measurement. B-mode ultrasonography was carried out successfully in the claws of five living cattle.     

Geometric symmetry of the solar surface of hooves of thoroughbred racehorses

OBJECTIVE: To define a 3-dimensional (3-D) coordinate system with clear definitions of origins and axes relative to hoof anatomic features and determine whether solar surfaces of Thoroughbred racehorse hooves have geometric asymmetry in the mediolateral and dorsopalmar directions. SAMPLE POPULATION: Left forelimb hooves from 20 Thoroughbred racehorse cadavers. PROCEDURE: A right-handed 3-D coordinate axes system centered on the collateral sulci was defined for the left front hoof. Orthogonal distances of anatomic features from the dorsopalmar axis and the plane coincident with the ground were measured and compared between medial and lateral sides and between dorsal and palmar regions of the hoof. RESULTS: The hoof was wider and had a greater radius laterally than medially. The most distal part of the lateral bar of the frog was further from the dorsopalmar axis than that of the medial bar. Overall, mediolateral asymmetries in depth were not observed. The sole at the perimeter was deeper medially in the dorsal part of the hoof and laterally in the palmar part, with depth overall being greater palmarly than dorsally. Most features had dorsopalmar asymmetry. CONCLUSIONS AND CLINICAL RELEVANCE: When the angle bisected by the collateral sulci is used to determine the dorsopalmar axis of the hoof, most central structures (bars and collateral sulci) have mediolateral symmetry. However, the hoof wall and sole have some mediolateral asymmetries and most structures have dorsopalmar asymmetry. These findings may assist the development of devices for attachment to hooves and studies of the interaction of hooves with bearing surfaces.     

Elastic properties of hoof horn on different positions in the bovine claw

Hind claws of 15 adult, sound Fleckvieh cows were used for material analysis. The elastic modulus was tested in tension tests according to EN ISO 527 and ASTM D 638-03 at a universal material testing machine. Samples were taken from different segments of the bovine claw to find the differences in material properties. Samples orientation was parallel to the horn tubules and transversal, respectively. Dry matter of the test samples was determined at the time of testing. Elastic modulus values were highest with mean = 659.7 N/mm2 at the dorsal wall. Values dropped axial to 416.3 N/mm2, abaxial to 343.9 N/mm2 for longitudinal (parallel) samples and to 433.1 N/mm2 for transversal samples. The elastic modulus of the sole segment was found to be 172.1 N/mm2. No difference was calculated neither between right and left feet, lateral and medial claws, nor between longitudinal and transversal samples.     

Characterisation of white line degeneration in sheep and evidence for genetic influences on its occurrence

Shelly hoof in sheep occurs when the hoof wall becomes detached from the laminar corium; it often then becomes impacted with debris leading to infection, pain and lameness. The problem of shelly hoof is under-reported and is often confused with classical footrot. A study was conducted using data on 9,169 Blackface and Texel sheep from 22 farms in the UK. The aim of the study was to estimate the prevalence of shelly hoof in these breeds, to quantify the genetic basis to shelly hoof, and to describe a pilot trail to characterise the physical properties of horn using Transmission Electron Microscopy (TEM). The results showed that shelly hoof has a high prevalence (47% for Blackface and 24% for Texel ewes) and is under moderate genetic control (h² = 0.3). The TEM images showed fragmented dorsal wall horn, microscopic crevices leading deeper into the hoof accompanied by bacteria and other micro-organisms. The invasion of bacteria was facilitated by separation of the cells in a characteristic ‘un-zipping’ phenomenon, poor attachment of the keratin to the cell membranes, poor cell membrane quality with a ‘lace-chain’ effect and incomplete keratinisation of the cells.     

The effect of flat horseshoes, raised heels and lowered heels on the biomechanics of the equine hoof assessed by finite element analysis (FEA)

The biomechanical effects of lowering and raising the heels were studied using a finite element (FE) computer model of the equine hoof capsule consisting of 18,635 finite elements. A static load of 3000 N was distributed to nodes of the inner hoof wall (80%) according to the suspension of the coffin bone, 20% loaded sole and frog. When loaded the FE hoof capsules showed the following deformations: the proximal dorsal wall moves back, the quarters flare to the side and sole and frog perform a downward movement. Stresses are high in the material surrounding the quarter nails, in the heels and in the proximal dorsal wall. Three types of horseshoes were simulated, a regular shoe with flat branches, a shoe with 5 degrees raised heels and a shoe with 5 degrees lowered heels. Raising the heels resulted in significantly (P < 0.05) low stress and displacement values. The lowered heels model calculated highest stress and displacement values and the results of the FE model with the regular horseshoe were found in between.     

马、骡蹄角质某些物理性能的测定

对马(20)、骡(10)蹄角质的测定结果表明,硬度、弹性,抗张强度、弹性模量均为蹄壁>蹄底>蹄叉;蹄底的磨耗大于蹄壁;蹄壁的磨擦系数大于蹄底;蹄壁纵向拉伸的抗张强度大于其横向拉伸,强度越大其伸长率越小;在相同压力下,蹄壁侧压弹性模量值大于纵压值,而蹄底侧压模量值则小于纵压值;蹄角质在材料力学上表现为一种各向异性的特点。作者认为,马、骡蹄角质物理性能的差异与蹄病的发生密切相关。     

Zur mikroskopischen Anatomie der Epidermis an der Schweineklaue

To gain a basis for diagnosing the frequent pathological changes of the porcine hoof, the fine structure of normal hoofs taken from landrace pigs was examined with the light microscope, the polarizing microscope and the electron microscope. The hoofs were taken mostly from 6 to 7 month-old slaughtered pigs, but also from younger animals, beginning at birth. The hoofs destined for light microscopical study were not softened before frozen sections were made; these sections were stained while floating on the stain. The hoof horn of the pig consists of mostly flat cells which are shaped like pancakes. The horn tubules often have oval cross sections and do not present a distinct boundary with the adjacent intertubular horn. The harder horn of the hoof has more horn tubules per mm² than the softer horn.     

Ultrasonographic and macroscopic comparison of the thickness of the capsule,corium, and soft tissues in bovine claws: an in vitro study

This study aimed to compare thickness of the capsule, corium, and soft tissues measured ultrasonographically and macroscopically in selected regions of bovine claws. A hundred and twenty claws (n = 120) of 15 healthy Holstein bovines were obtained. After cleaning the claws, ultrasonographic measurement of the capsule, corium, and soft tissues was performed while submerging the claws in a water bath. Macroscopic measurements were taken after cutting of the claws axially. These values were compared statistically. According to the macroscopic measurements, the mean thickness ± standard deviation (SD) of the capsule for dorsal wall and sole was 6.2 ± 0.1 and 9.5 ± 0.4 mm, respectively. The thickness of the corium and soft tissues for dorsal wall and sole was 4.5 ± 0.1 and 5.3 ± 0.1 mm, respectively. Ultrasonographically, the mean thickness ± SD of the capsule for dorsal wall and sole was 4.7 ± 0.1 and 7.8 ± 0.3 mm, respectively. The thickness of the corium and soft tissues for dorsal wall and sole was 4.3 ± 0.1 and 5.9 ± 0.2 mm, respectively. Findings demonstrated that ultrasonography can be reliably to measure of the thickness of the hoof capsule, corium, and soft tissue in bovine claw.     

A macroscopic study of the vascular system of the bovine hind limb claw

The vascular organisation of the hind limb claws from clinically normal cattle was studied using plastic corrosion casts. The proper digital artery ran dorsodistally within the interdigital space and gave off a bulbar branch to the bulb, a plantar branch to the axial aspect of the wall and sole, and a coronary artery to the coronary margin and proximal wall. The artery entered the distal phalanx at the axial foramen, coursed through the nutrient canal as the terminal arch, and emerged at the abaxial foramen to anastomose with branches of the bulbar artery. During its interosseous course, the terminal branch gave off 6-8 primary branches which provided the arterial supply to the coronary margin, axial and abaxial wall, white zone and sole. The dorsal proper digital vein and axial and abaxial proper plantar digital veins drained all venous blood from the claw. Each of these large veins arose from numerous smaller veins which joined to form the venous networks of the coronary margin, wall, sole and bulb. A large arched vein passed through the coronary cushion and formed a connection between the superficial coronary venous plexus and the intra-osseous plexus of the distal phalanx. The axial and abaxial proper plantar digital veins were connected by a vessel through the bulbar cushion. At the same level, the axial proper plantar digital veins of the lateral and medial claw were connected to each other by a large vessel through the interdigital space, situated just proximal to the distal interdigital ligaments. This resulted in the presence of a complete distal venous arch along the plantar aspects of both claws. The parietal collecting veins drained most of the venous blood from the sole, white zone and wall, and connected directly to the venous plexuses of the coronary margin and bulb. Valves occurred commonly in medium and larger calibre veins throughout the claw.     

Decreased expression of p63, a regulator of epidermal stem cells, in the chronic laminitic equine hoof

Reasons for performing study: Abnormal epidermal stem cell regulation may contribute to the pathogenesis of equine chronic laminitis. Objective: To analyse the involvement of p63, a regulator of epidermal stem cell proliferative potential, in chronic laminitis. Methods: Epidermal tissues from skin, coronet and lamellae of the dorsal foot were harvested from 5 horses with chronic laminitis and 5 control horses. Tissues were analysed using histopathology, immunofluorescence microscopy and quantitative immunoblotting Results: Hoof lamellae of laminitic horses had a lower frequency of p63 positive cells than control lamellae, particularly in the distal region. Quantitative immunoblotting confirmed reduced p63 expression in the laminitic distal lamellar region. The decreased p63 expression in laminitic epidermal lamellae was most apparent in the abaxial region adjacent to the hoof wall and highly associated with the formation of terminally differentiated, dysplastic and hyperkeratotic epidermis in this region, whereas lamellae from control horses maintained high p63 expression throughout the axial‐abaxial axis. Conclusions: Expression of p63 in equine skin resembles that reported in other species, including man and rodents, suggesting that p63 can serve as a marker for the proliferative potential of equine epidermal stem cells. p63 expression was significantly lower in the chronic laminitic hoof than in that of control horses, suggesting laminitic hoof epithelium has more limited proliferative potential with a shift towards differentiation. This may reflect reduced activity of epidermal stem cells in laminitic hoof. It is proposed that p63 contributes to the maintenance of hoof lamellae and that misregulation of p63 expression may lead to epidermal dysplasia during lamellar wedge formation. Potential relevance: This study suggests that loss of epidermal stem cells contributes to the pathogenesis of equine laminitis. Autologous transplantation of p63‐positive epidermal stem cells from unaffected regions may have regenerative therapeutic potential for laminitic horses.     

Keratomas in horses: seven cases (1975-1986)

The diagnosis of keratoma in 7 horses and their treatment and outcome were evaluated. Horses were 2 to 20 years old, of various breeds, and were intact or castrated males. All were lame, and 6 horses had had previous injuries of the affected hoof that had not responded to prior treatments. Only 1 hoof was affected in each horse. Keratomas were beneath the hoof wall (6 horses) or sole (1 horse). Radiographically, a circular or semicircular defect with a discrete margin was present in the distal portion of the third phalanx in 3 horses. Grossly, keratomas were firm solitary masses (1.5 to 5 cm diameter); gray, tan, or yellow; and oval or conical. Keratomas were excised from beneath the hoof wall by removing a section of hoof wall (5 horses) or by undermining the wall beginning at its junction with the sole (1 horse). A keratoma beneath the sole in 1 horse was excised by excavating the sole to the level of the palmar surface of the third phalanx. Keratoma was verified microscopically by the presence of characteristic rings of squamous epithelial cells with abundant keratin. Purulent exudate and inflammatory infiltrate often were present concurrently, reflecting an associated localized infection. Aftercare included daily application of an antiseptic iodine solution and foot bandages, and shoeing with a treatment plate several weeks after surgery. Hoof and sole defects healed completely between 6 months and 1 year. By 1 year after surgery, 6 horses were sound, and keratoma had not recurred.     

A method of biomechanical testing the suspensory apparatus of the third phalanx in cattle: a technical note

There are a number of differences between the claws of the front and hind limbs in cattle concerning macroscopic shape, chemical and physical properties of claw horn and epidemiological data. Front claws have superior data in almost every respect. Virtually no information is available on the mechanical stability of the suspensory apparatus of the third phalanx. It was the objective of this study to develop a method to measure the maximum strength (N/mm2) of the suspensory apparatus of the bovine distal phalanx. This apparatus includes all layers of tissue such as bone, corium, corioepidermal junction and wall horn. The feet of 13 clinically healthy beef bulls were collected from the local abattoir. Testing was performed in three locations of the wall segment (dorsal, abaxial, axial) in a material testing machine. The corium and the corioepidermal junction were set under increasing tension until failure occurred. Maximum stress was recorded. The values ranged between 2.47-3.13 N/mm2 (dorsal), 4.08-4.87 N/mm2 (abaxial) and 2.27-2.66 N/mm2 (axial). No differences could be validated statistically between front and hind claws and between lateral and medial claws respectively. Abaxial ultimate stress values were significantly higher than dorsal and axial ones. Between the two a statistical difference could not be validated. Most of the specimens (57.1%) were torn apart at the corioepidermal junction, almost a quarter of the samples (27.6%) were to a degree disrupted at the corioepidermal junction and to a certain extent in the deeper layers of the corium. A few samples showed disruption within the horn (11.5%) or at the attachment of the third phalanx (3.8%). Biomechanical features of the experimental set-up are discussed. The biological significance of these findings is that the body weight and the additional dynamic loading is predominantly taken over by the abaxial part of the claw, while the dorsal and axial aspects are less loaded.