Behavior during transition differs for cows diagnosed with claw horn lesions in mid lactation

Claw horn lesions, including sole hemorrhages and sole ulcers, are a major cause of lameness in dairy cattle. These lesions often develop in the weeks around calving and become visible 8 to 12 wk later. The aim was to determine whether cows that are diagnosed with claw horn lesions several weeks after calving behave differently during the calving period when the lesions are thought to first develop. The claws of 26 multiparous Holstein dairy cows were scored for sole hemorrhage severity and presence of sole ulcers 2 wk before calving, 3 wk after calving, and every 4 wk thereafter until 15 wk after calving. Of the 26 cows, 13 cows had no or low lesion scores up to 15 wk and 13 cows had at least 1 new severe lesion or sole ulcer diagnosed between 7 and 15 wk after calving. Behavior (dry matter intake, feeding time and rate, number and size of meals, standing time, number and duration of standing bouts, location of standing, and displacements at the feed bunk) was recorded from 2 wk before calving to 3 wk after calving. Behavior during the 2 wk before calving, first 24 h after calving, 1 wk after calving, and 2 to 3 wk after calving was compared between cows with and without lesions in mid lactation. Multivariate regression was used to determine the predictive value of each behavior and combination of behaviors on lesion diagnosis. Cows diagnosed with lesions in mid lactation spent more time standing than cows without lesions during the 2 wk before (832 ± 29 vs. 711 ± 29 min/d) and 24 h after (935 ± 46 vs. 693 ± 46 min/d) calving. These differences were driven by an increase in the time spent perching with front feet in the stall (241 ± 22 vs. 147 ± 22 min/d at 2 wk before calving) and an increase in standing bout duration (101 ± 10 vs. 56 ± 10 min/bout at 24 h after calving). Compared with cows without lesions, cows with lesions consumed feed at a faster rate (86 ± 3 vs. 77 ± 3 g/min) during the 2 wk before calving and consumed more feed (17.9 ± 0.9 vs. 12.3 ± 0.9 kg/d) during the 24 h after calving. The number of displacements at the feeder was not different between groups. These results indicate that a combination of feeding and standing behavior during the transition can serve as early indicators of claw horn lesions in mid lactation.     

Reduced test-day milk fat percentage in cows diagnosed with claw horn lesions during routine claw trimming

The objective of this study was to investigate whether cows diagnosed with claw horn lesions during routine claw trimming had an altered milk fat percentage on test days preceding the diagnosis, compared with cows without such lesions. Data included date of trimming, cow number, type, and location of claw lesions recorded by a professional claw trimmer during routine trimming sessions on 11 commercial dairy farms, and monthly test-day data. Recordings from 4,381 cows were included in the analysis. Three hundred seventy-two (8.5%) cows were diagnosed with at least one claw lesion. One hundred ninety-five (4.4%) cows were classified as cases of laminitis and 169 cows (3.9%) were classified as cases of infectious claw diseases. Average test-day milk fat percentage was estimated from monthly test-day data using a mixed model with a random farm effect and a marginal effect for repeated measurements. Estimated milk fat percentage of cows classified as cases of laminitis on the day of claw trimming was 0.17 and 0.22% less than that of cows with either no claw lesion or those with an infectious claw disease, on test days occurring 2 and 5 mo before claw trimming, respectively. To the best of our knowledge, decreased test-day milk fat percentage in dairy cows diagnosed with claw horn lesions has not been described. Because decreased milk fat and claw horn lesions have both been associated with subacute ruminal acidosis, it is plausible that subacute ruminal acidosis played a role in the association we found. A prospective study with a greater frequency of claw examinations would be necessary to help clarify these associations. Our findings indicate that test-day milk fat might be useful as a tool for monitoring cows and farms at risk for claw horn lesions.     

Distribution of bovine milk sialoglycoconjugates during lactation

The sialoglycoconjugate content of human milk has been extensively studied. However, little attention has been paid to the changes occurring in these compounds in bovine milk during lactation. Since sialoglycoconjugates are very abundant in milk from the early stages of lactation, they have been suggested to be important for the nutrition of the newborn during the first months of life. The distribution of sialoglycoconjugates (expressed as glycoconjugate-bound sialic acid) from four different stages of lactation (colostrum, transitional, mature, and late-lactation milks) was investigated in four Spanish-Brown cows. All the fractions studied (total sialic acids, glycoproteins, oligosaccharides, casein, and gangliosides) showed a similar trend. We found the highest values in the colostrum, these decreasing in transitional and mature milks and increasing again in late-lactation milk. We also found a selective change in the relative contents of glycoprotein- and oligosaccharide-bound sialic acids. In mature milk, the latter increased up to 80% (59% in colostrum) and the former decreased to 3.9% (35.3% in colostrum). It would appear that the decrease in oligosaccharide-bound sialic acid is compensated by the increase in glycoprotein-bound sialic acid. From these results, it is deduced that newborn infants or calves fed with infant formulas or milk replacers, respectively, should be supplemented with sialoglycoconjugates to approximate the composition of human and cow milk as far as is practicable.     

Visceral tissue growth and proliferation during the bovine lactation cycle

Twenty one multiparous, nonpregnant, lactating dairy cows were used to assess the impact of stage of lactation on visceral tissue mass and small intestinal cell proliferation. Cows were slaughtered at each of 4 stages of lactation: 14, 90, 120, and 240 d of lactation. With stage of lactation, DMI increased through d 90 and thereafter remained similar through d 240 (quadratic). Carcass weight and empty body weight (EBW) declined with stage of lactation through d 120 and increased thereafter (quadratic). As a percentage of EBW, rumen, small intestine, and liver weights increased with increasing stage of lactation (quadratic), increasing from 14 to 120 d and declining through 240 d. Stage of lactation did not have a measurable affect on reticulum, omasum, abomasum, or large intestine weights as a percentage of EBW. Visceral adipose mass as a percentage of EBW declined with stage of lactation to a minimum at 120 d and increased by 240 d (quadratic). Concentrations of RNA and DNA of digestive tract organs were largely unaffected by stage of lactation with the exception of the liver DNA concentration through d 120 (quadratic). The proliferative growth fraction (Ki67) was unaffected by stage of lactation. However, bromo-deoxyuridine labeling of jejunal crypts exhibited a cubic response with stage of lactation and tritiated thymidine incorporation by duodenal epithelium increased with stage of lactation through d 120, declining thereafter (quadratic). Mass of visceral tissues increase to meet the energetic demands of lactation and that increased absorption capacity of the intestines is achieved by hyperplastic growth of the intestinal epithelium.     

The pressure distribution under the bovine claw during square standing on a flat substrate

The distribution pattern of pressure over the bovine claw was investigated to test the hypothesis that the ground reaction force is unevenly distributed and makes some regions of the claw more prone to overloading and injury than others. In eight recently trimmed Holstein Friesian cows, the distribution of vertical pressure was measured during square standing with a spatial resolution of 2.6 sensors/cm2 and a temporal resolution of 313 measurements/s. In each animal, the localization of maximum pressure per foot and per claw was determined during five trials. In the front limb, maximum pressures were normally found on the medial claw; in the hindlimb they were located on the lateral claw. In both claws, the highest pressures were found on the sole of the foot and not on the wall. In the front limbs, maximum pressures were located in the posterior portion of the sole; in the hind limb in the anterior portion. There was no difference in the location of the maximum pressure between the medial and lateral claw in either limb. The regions in which these maximum pressures occur are known to be relatively susceptible to injuries. This could indicate a causal relation between the location of pressure concentrations and claw diseases found in clinical observations.     

The effect of claw horn disruption lesions and body condition score at dry-off on survivability, reproductive performance, and milk production in the subsequent lactation

The objective of this study was to evaluate the effects of claw horn disruption lesions (CHDL; sole ulcers and white line disease) and body condition score (BCS) at dry-off on survivability, milk production, and reproductive performance during the subsequent lactation. An observational prospective cohort study was conducted on a large commercial dairy in Cayuga County, New York, from September 2008 until January 2009. A total of 573 cows enrolled at dry-off were scored for body condition and hoof trimmed; digits were visually inspected for the presence of CHDL. The BCS data were recategorized into a 3-level variable BCS group (BCSG), with cows with BCS <3 placed in BCSG 1 (n = 113), cows with BCS = 3 placed in BCSG 2 (n = 254), and cows with BCS >3 placed in BCSG 3 (n = 206). Cows in BCSG 2 were 1.35 and 1.02 times more likely to conceive than cows in BCSG 1 and 3, respectively. The cull/death hazard for BCSG 1 cows was 1.55 and 1.47 times higher than for cows in BCSG 2 and BCSG 3, respectively. Milk yield for cows in BCSG 2 (44.6 kg/d, 95% CI 43.4-45.8) was significantly greater than that for cows in BCSG 1 (41.5 kg/d, 95% CI 39.8-43.3). Cows with previous lactation days open ≤91 had 1.6 times higher odds of being classified into BCSG 1 at dry-off; cows with previous lactation mature-equivalent 305-d milk >14,054 kg had a similar 1.6 times higher odds of being classified into BCSG 1. Claw horn disruption lesions were found in 24.4% of the cows (n = 140) at dry-off. Cows without CHDL were 1.4 times more likely to conceive than cows with CHDL. Additionally, lesion cows were 1.7 times more likely to die or be culled than nonlesion cows. Absence of CHDL did not have a significant effect on milk yield. These findings highlight the importance of claw health and BCS at the end of lactation on future survival and performance.     

Prevalence and ultrastructural characteristics of bovine mammary corpora amylacea during the lactation cycle

Morphologic and histochemical study of spherical, lamellated inclusion bodies in bovine mammary parenchyma established presence of corpora amylacea. The majority (90%) were in alveolar lumina, followed by stroma (7%) and epithelium (3%). Occurrence of corpora amylacea was unrelated to lactation age, intramammary infection status, milk somatic cell concentration, or milk production. Quantitative analysis demonstrated a progressive increase of prevalence of these structures from the early to later stages of lactation, followed by a return during the late dry period to concentrations at early lactation. Ultrastructural study revealed deeply basophilic, dense, lamellated deposits (70%) and less dense, fibrillar deposits (30%). Dense bodies stained partially or not at all for amyloid but stained for presence of calcium salts, whereas fibrillar bodies were amyloid positive and calcium negative. Corpora amylacea were most prevalent in fully differentiated parenchymal areas (47.1 bodies/unit tissue area of 2.4 X 10(6) micron 2), less numerous in intermediately differentiated areas (23.4), and sparse in nonsecretory areas (7.1). Morphologic relationships between deposits and parenchyma suggest that corpora amylacea suppress milk secretion and flow during late lactation in isolated areas by engorging luminal spaces and clogging small ducts, leading to milk stasis and involution.     

Changes in physical properties of bovine milk from the colostrum period to early lactation

The aim of this study was to analyze individual cows’ samples from the colostrum, postcolostrum, and early lactation periods to investigate how milk composition, physical properties, stability, and suitability for processing change throughout this period. Attention was paid to the first week postpartum in which the composition of bovine mammary secretion can change markedly. Properties including pH, titratable acidity, ethanol stability (ES), rennet clotting time, and casein micelle size were analyzed, together with some compositional factors such as fat, total protein, lactose, total and ionic calcium, magnesium, citrate, phosphorus, sodium, and potassium. Total Ca (36.2 mM) and free ionic Ca (2.58 mM), Mg (5.9 mM), P (32.2 mM), and Na (24.1 mM) appeared to be high on d 5 postpartum, having decreased substantially over the first 5 d; they gradually decreased thereafter. The average pH on d 5 was only 6.49, compared with 6.64 at 1 mo postpartum. Stability measurements showed that the average ES on d 5 was 70% and the rennet clotting time was 12.2 min, which were significantly lower than values at later stages. A number of milk properties including ES, pH, protein content, and Ca²⁺ concentration could be useful for identifying the point of transition from colostrum to the early lactation period. Knowing the composition and physical properties of colostrum and postcolostrum secretions will help establish when such milk is suitable for processing and determine the best use for that milk.     

Concentrations of sialyloligosaccharides in bovine colostrum and milk during the prepartum and early lactation

Sialyloligosaccharides and sialylglycoconjugates in colostrum and milk are regarded to be important biological components with respect to be source of brain gangliosides in infant and to be antiinfectional components for the attack by the pathogenic bacteria and virus. Several acidic oligosaccharides have been characterised in both bovine and human milk or colostrum. The sialyloligosaccharide content of human colostrum and milk has been extensively studied, whereas that of cows milk and colostrum has received less attention. In this study, the concentrations of three sialyloligosaccharides of bovine colostrum and milk were determined at various stages during the prepartum and the first 7 d postpartum. The concentration of 3'SL (Neu5Ac(alpha2-3)Gal(beta1-4)Glc) reached a maximum value of 0.85 mg/ml immediately following parturition while the concentrations of 6'SL (Neu5Ac(alpha2-6)Gal(beta1-4)Glc) and 6'SLN (Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc) of 0.14 and 0.12 mg/ml, respectively, were much lower at this initial stage, although these concentration were maximum immediately following parturition. Bovine colostrum, especially that collected immediately after parturition, may be suitable as a source of 3'SL and other sialyloligosaccharides for use as additives by the food or pharmaceutical industries.     

Differential distribution of T lymphocyte subpopulations in the bovine mammary gland during lactation.

The existence and distribution of T lymphocyte subpopulations in the mammary parenchymal tissue of cows were immunohistochemically detailed during early lactation. CD2+, CD4+, and CD8+ T lymphocytes were localized primarily in the mammary parenchymal tissue. CD8+ T lymphocytes were predominant over CD4+ T lymphocytes and occurred in close contact with the alveolar epithelium and between epithelial cells in the central area of the upper mammary gland. CD4+ T lymphocytes were present in equal numbers in the epithelial and connective tissue area. Occasionally, both CD4+ and CD8+ T lymphocytes formed cell clusters in the interalveolar connective tissue. The ratio of CD4+ T lymphocytes to CD8+ T lymphocytes was less than 1.0 and was lower in the epithelial area than in the connective tissue. The distribution of CD4+ and CD8+ T lymphocytes was similar in the parenchymal tissue of the gland cistern. The observation that there is a preferential presence of CD8+ T lymphocytes in the epithelial area of the bovine mammary gland during early lactation might indicate that these cells participate in the maintenance of the integrity of the epithelium.     

Age, segment, and horn disease affect expression of cytokines, growth factors, and receptors in the epidermis and dermis of the bovine claw

The aim of this study was to examine changes in RNA expression for growth factors, cytokines, and receptors in epidermal-dermal tissues of the bovine claw relative to host age, claw segment, and disease state of the horn. Epidermal-dermal tissues were collected from the coronary, wall, sole, and bulb segments of 8- to 9-mo-old Holstein fetuses, normal adult cows, and adult cows with sole ulceration. Anatomic and pathologic characteristics were determined in tissues stained with eosin and hematoxylin, and RNA expression levels were evaluated using real-time, quantitative PCR. In normal tissues, certain RNA expression levels were clearly affected by host age: 290.0-, 610.0-, 53.4-, and 8.1-fold greater expression of granulocyte-macrophage colony stimulating factor was observed in fetal coronary, wall, sole, and bulb segment relative to adult tissues, respectively. A claw segment effect was also observed in that IL-1α expression was greater (1.59-fold) in the normal adult wall relative to the coronary segment, and IL-18 expression was greater (16.2-fold) in the normal adult sole compared with the coronary segment and 2.88 greater in the fetal sole relative to the bulb segment. Sole ulceration was associated with hemorrhage, thrombosis, inflammation, and striking increases in IL-1β, IL-18, and inducible nitric oxide synthase, and with less dramatic, albeit measurable, changes in IL-1 type I receptor, IL-1 receptor antagonist, and tumor necrosis factor-α. Amidst striking increases in keratinocyte growth factor receptor (i.e., 21.0-fold, 10.4-fold, 0, and 21.6-fold in the coronary, wall, sole, and bulb segments, respectively), a concomitant decrease occurred in keratinocyte growth factor (i.e., 0.80-, 0.54-, 0.56-, and 0.72-fold, respectively). The results demonstrated changes in disease state and, to a lesser extent, claw segment and were accompanied by alterations in the RNA expression of several cytokines, growth factors, and receptors present in the normal claw.     

The effects of social environment on standing behavior and the development of claw horn lesions

The primary aim of this prospective experimental study was to evaluate how the social environment after calving influenced standing behavior in primiparous cows. At calving, primiparous cows were mixed with familiar peers in a low-stocked pen (≤33% stocking density; n = 22) or mixed with unknown older cows at 100% stocking density (n = 20). All study cows were mixed with older cows 3 wk after calving. Time spent standing and perching (standing with only the front feet in the stall) were measured d 1 to 3 after calving using 5-min scan sampling. To evaluate if the low-stocked treatment constituted a low-stress social environment, agonistic interactions at the feed barrier were measured for 90 min following feed delivery for a subsample of cows in both treatments (12 cows/treatment). The daily behavioral time budget, including the 90 min following milking, was examined for this subset of cows. A secondary aim was to assess if the social environment after calving was related to the risk of developing claw horn lesions later in lactation. Sole and white line lesions were recorded at wk 6 and 12 after calving, and cows were categorized as either having or not having at least 1 hemorrhage of severity ≥3 (scale 1 to 5) for each lesion type and assessment. Prolonged standing after regrouping was not observed, and we found no differences in standing time and time spent perching between treatments. Agonistic behaviors directed toward the focal cows occurred less frequently in the low-stocked pen compared with the control. The number and severity of sole and white line lesions increased after calving. At wk 6 postpartum there was a numeric (but not statistically significant) difference between treatments in the proportion of primiparous cows that had white line hemorrhages of severity score ≥3 (low-stress social environment: 20% vs. control: 50%). In conclusion, under the conditions of this study the social environment did not influence standing behavior, but did affect agonistic interactions and may have influenced the risk of claw horn lesions in the weeks following calving. Further studies should evaluate the relationship between the social environment and claw health.     

Regulation of bovine adipose tissue metabolism during lactation. 4. Dose-responsiveness to epinephrine as altered by stage of lactation

Adaptations in adipose tissue lipolysis responsiveness to doses of epinephrine were determined in first lactation Holstein cows. Subcutaneous adipose tissue was biopsied at -30, -15, 30, 60, 120, 180, and 240 d about first calving. Glycerol and fatty acid release from tissue triglycerides were determined in vitro in the presence of 10(-8) to 10(-4) M epinephrine. Basal lipolysis increased postpartum and remained elevated through 240 d of lactation. Glycerol release in response to graded doses of epinephrine increased from 30 d prepartum to 30 d postpartum and remained elevated through 240 d. The highest net response was reached at 120 d and was maintained to 240 d. Increases during lactation were noted in actual glycerol release, net response (stimulated minus basal activity), and maximum net response (calculated from reciprocal plots). Maximal and submaximal response of fatty acid release to epinephrine increased post partum with maximal adaptation occurring by 30 d and remaining elevated through 240 d. Maximum net response of glycerol release at 30 d was related positively (r = .73) to milk energy secretion and negatively to energy intake (r = -.57) and energy balance (r = -.79). Net maximum free fatty acid response at 120 d related positively (r = .89) to milk energy secretion and negatively (r = -.81) to energy balance. The epinephrine responsiveness of adipose tissue increases during lactation in a manner consistent with whole body energy inputs and outputs.     

Efficacy of chlorhexidine as a postmilking teat disinfectant for the prevention of bovine mastitis during lactation

A natural exposure trial was conducted for 12 mo in a herd of 150 lactating Jersey cows to determine efficacy of a .35% chlorhexidine teat dip containing a glycerine emollient for the prevention of bovine intramammary infections. Right teats of cows were dipped in the experimental teat dip after milking machine removal and left teats were not dipped. The herd was free of Streptococcus agalactiae and had a low prevalence of Staphylococcus aureus. Most new major pathogen intramammary infections resulted from Streptococcus species, primarily Streptococcus uberis and Streptococcus dysgalactiae. New infections by Streptococcus species were significantly lower in teats dipped in chlorhexidine than in undipped teats. Overall efficacy of the chlorhexidine teat dip against major mastitis pathogens was 50%. The experimental teat dip also reduced coagulase-negative Staphylococcus species infections 49.0% and Corynebacterium bovis infections 65.2%. Overall efficacy against minor mastitis pathogens was 54.0%. No irritation or chapping of teats dipped in the experimental teat dip was observed.     

Technical note: Effects of frozen storage on the mechanical properties of the suspensory tissue in the bovine claw

It is proposed that a softening of the suspensory tissue in the claw is involved in the development of lameness and claw lesions in cattle. A relatively small amount of research has been carried out to verify this theory. Research in this area would be simplified if mechanical testing of the suspensory tissue could be performed on frozen and stored specimens. The current study tested whether freezing of the specimens changes the suspensory tissues’ mechanical properties. Limbs from 3 freshly slaughtered Danish Holstein dairy cows and 6 nonpregnant Angus heifers, without clinical signs of lameness, were allocated to 1 of 2 treatments (frozen or nonfrozen) in such a way that each cow was represented in each treatment group with a frozen limb and a corresponding nonfrozen limb (i.e., frozen left front, fresh right front, and so on). The frozen limbs were kept at −18°C for a week before processing and the nonfrozen limbs were processed within 2 h of slaughter. Two samples measuring 8 × 8 mm were cut from the abaxial side of each claw in such a way that the sample included the horn of the abaxial wall, pedal bone, and the interposed corium. The samples were kept on ice until being mounted in a large deformation rheometer with an extension testing frame, fixed by the horn and the pedal bone, and loaded to failure. During deformation force and displacement data were recorded, from which corresponding stress and strain were calculated. Young's modulus (a measure of tissue elasticity or stiffness) and a measure of physiological support (PS; force needed to displace the sample 1 mm) were calculated from the data. The response variables, Young's modulus and PS, were analyzed separately by a mixed model. The explanatory variables were treatment (frozen or nonfrozen), limb (front or back), claw (medial or lateral), position of the sample (dorsal or palmar-plantar), and group (Angus or Holstein). Interactions between group and treatment and between limb, claw, and sample position were included in the model. Cow identity was included as a random effect. Model reduction was performed by stepwise backward elimination, until all remaining terms were significant at the 5% level or less. Freezing had no effect on the elasticity of the suspensory apparatus or on PS. However, PS was affected by limb (hind legs had higher PS values than front) and the position of the sample (palmar-plantar samples had higher PS values than dorsal). The Angus group had higher PS values than the Holstein group, but the groups differed in age, parity, body weight, lactation, housing, and management, as well as in breed; therefore, further studies are needed to investigate these effects. The results indicate that mechanical testing of bovine claw suspensory tissue can be performed on specimens that have been frozen, thus aiding research in the mechanical aspect of bovine lameness and claw lesions.     

Glycosidase activities in bovine milk over lactation

The presence and activity of glycosidases in Irish bovine milk over three months of lactation was investigated. A low level of variation between animals was observed and the highest level of glycosidase activity was present in colostrum, decreasing through transitional milk production to minimal but constant levels in mature milk. N-acetyl-β-d-glucosaminidase, α-l-fucosidase, α-galactosidase and N-acetyl-neuraminidase appear to be the most biologically relevant glycosidases in bovine milk. The elevated levels of enzymatic activity in colostrum suggests the milk glycosidases may play a role in the digestion of bovine milk glycans in the infant mammal, possibly acting as substitutes for bacterial glycosidases prior to colonisation by the gut microflora, which are involved in in vivo oligosaccharide metabolism in the lower gastrointestinal tract.     

Invited review: formation of keratins in the bovine claw: roles of hormones, minerals, and vitamins in functional claw integrity

Keratins are the characteristic structural proteins of the highly cornified epidermis of the skin, feathers, and hoof. Keratin proteins provide the structural basis for the unique properties of the biomaterial horn and its protective function against a wide range of environmental factors. Hoof horn is produced through a complex process of differentiation (keratinization) of epidermal cells. Formation and biochemical binding of keratin proteins and synthesis and exocytosis of intercellular cementing substance (ICS) are the hallmarks of keratinization. It is finalized by the programmed death of the living epidermal cells, i.e., cornification, that turns the living epidermal cells into dead horn cells. The latter become connected by the intercellular cementing substance. The functional integrity of hoof horn essentially depends on a proper differentiation, i.e., keratinization of hoof epidermal cells. Keratinization of hoof epidermis is controlled and modulated by a variety of bioactive molecules and hormones. This process is dependent on an appropriate supply of nutrients, including vitamins, minerals, and trace elements. Regulation and control of differentiation and nutrient flow to the epidermal cells play a central role in determining the quality and, consequently, the functional integrity of hoof horn. Decreasing nutrient supply to keratinizing epidermal cells leads to horn production of inferior quality and increased susceptibility to chemical, physical, or microbial damage from the environment. A growing body of evidence suggests that hormones, vitamins, minerals, and trace elements play critical roles in the normal development of claw horn and correct keratin formation.     

Synthesis and distribution of cytokeratins in healthy and ulcerated bovine claw epidermis

Keratinization of the epidermal cells of the bovine claw generates the horn that gives the tissue its mechanical strength. Disruption of keratinization is likely to have a detrimental effect on the strength and integrity of the horn, and could lead to solar lesions and lameness. As part of a wider investigation of the cell biological causes of lameness in dairy animals, we have compared keratin synthesis and distribution in healthy bovine claw tissue with those in hooves with solar ulcers. Protein synthesis was measured by [35S]-labelled amino acid incorporation in claw tissue explant cultures. [35S]-labelled protein synthesis was higher in tissue from diseased claws than in healthy claws, and highest at the ulcer site. The identity of proteins synthesised in vitro did not differ between healthy and diseased tissue. DNA synthesis indicative of cell proliferation was also elevated in diseased tissue. Immunoblotting after one- or two-dimensional electrophoresis showed cytokeratins (CK) 4, 5/6, 10 and 14 to be amongst those expressed in healthy claw tissue. The relative abundance of these keratins was not altered in healthy regions of ulcerated hooves, nor at the ulcer site, but CK16, not usually found in healthy tissue, was detected in the sole of diseased claws. CK5/6 and CK14 were shown by immunohistochemistry to be present in the basal epidermis of healthy tissue, whereas CK10 was found in supra-basal layers. In healthy tissue from ulcerated claws, this distribution was unaltered, but at the site of solar ulcers, CK5/6 and CK14 were each found in both basal and supra-basal epidermis. The study suggests that solar ulceration of the bovine claw is not associated with gross alteration in the keratin composition of the tissue, but causes abnormal distribution of cytokeratins, perhaps as a result of loss of positional cues from the basement membrane. Ulceration did, however, stimulate cell repair involving epidermal protein synthesis (including keratins), and keratinocyte proliferation.     

Biomechanics and histology of bovine claw suspensory tissue in early acute laminitis

Weakening of the suspensory tissue supporting the pedal bone is the central issue in the theory of acute bovine laminitis, but this aspect has never been tested. The objective of this study was to investigate the effect of laminitis on the suspensory tissue. The hypothesis was that clinical and histological signs of acute laminitis are associated with decreased strength of the suspensory tissue of the bovine claw. Nonpregnant dairy heifers (n = 10) received oral oligofructose overload (17 g/kg of body weight) and were killed 24 (n = 4) and 72 h (n = 6) after overload. Control heifers (n = 6) received tap water and were killed at 72 or 96 h. Clinical, orthopedic, and histological examinations were carried out to confirm the occurrence of laminitis. After euthanasia, 2 adjacent tissue samples including the horn wall, lamellar layer, dermis, and pedal bone were cut from the dorso-abaxial aspect of each claw. Tissue samples were kept on ice until mounted on a mechanical testing frame, fixed by horn and bone, and loaded to failure. A stress displacement curve was generated and measurements of physiological support (force needed to displace 1 mm beyond first resistance) and maximal support (force needed to break the tissue) were recorded. Heifers treated with oligofructose developed clinical signs consistent with ruminal and systemic acidosis after treatment as well as acute laminitis, characterized by weight shifting (35% of observations vs. 6% in controls), moderate lameness (100 vs. 17%, score of 3 out of 5 at 72 h), and reaction to hoof testing (30 and 50% at 48 and 72 h, respectively, vs. 0% in controls). Histological examination of claws from heifers killed 72 h after overload showed changes consistent with acute laminitis, including stretched lamellae, wider basal cells with low chromatin density, and a thick, wavy, and blurry appearance of the basement membrane. Biomechanical results showed no effect of oligofructose overload on physiological support of the suspensory tissue at 24 and 72 h after overload; in contrast, overload increased maximal support of the tissue 72 h after overload. Herd of origin and location of the sample had large effects on both physiological support and maximal support (herd = 547 N/cm²; location = 531 N/cm²) of claw suspensory tissue (herd = 260 N/cm²; location = 327 N/cm²). Despite clinical and histological signs of laminitis, no weakening of the suspensory tissue of the bovine claw was detected at 24 and 72 h after oligofructose overload. Herd factors appeared to be important for claw suspensory tissue strength.