Proteoglycan synthesis and content in articular cartilage and cartilage repair tissue in horses

Hexosamine concentration, DNA concentration, and [35S]sulfate incorporation for articular cartilage obtained from various sites in the metacarpophalangeal and carpal joints of horses were measured. The same measurements were made on the repair tissue filling full-thickness articular defects in the intermediate carpal bone and on cartilage surrounding partial-thickness defects 6 weeks after the defects were created arthroscopically. Cellularity (measured as DNA concentration), proteoglycan content (measured as hexosamine concentration), and proteoglycan synthesis (measured as [35S]sulfate incorporation) varied according to the site sampled. Cartilage from the transverse ridge of the head of the third metacarpal bone and the radial facet of the third carpal bone had the lowest hexosamine concentration, whereas rate of proteoglycan synthesis was lowest in cartilage from the transverse ridge of the head of the third metacarpal bone and the distal articular surface of the radial carpal bone. Repair tissue filling a full-thickness cartilage defect at 6 weeks was highly cellular. It was low in proteoglycan content, but was actively synthesizing these macromolecules. In contrast, the cartilage surrounding a partial-thickness defect was unchanged 6 weeks after the original defect was made.     

Low dietary silicon supplementation may not affect bone and cartilage in mature,sedentary horses

As osteoarthritis is a major cause of lameness in horses in the United States, improving collagen health prior to onset and increasing collagen turnover within affected joints could improve health- and welfare-related outcomes. Through its positive effects on bone mineral content and density and its role in increasing collagen synthesis, silicon (Si) may slow the development and progression of osteoarthritis, thereby reducing lameness. This study evaluated the hypothesis that Si supplementation would increase cartilage turnover through increased collagen degradation and formation markers, as well as bone formation markers, resulting in reduced lameness severity when compared with controls. Ten mature Standardbred geldings were assigned to either a Si-treated (SIL) or control (CON) group and group-housed on pasture for 84 d. Horses were individually fed to ensure no cross-contamination of Si other than what was present in the environment. For the duration of the study, SIL horses received a Si–collagen supplement at the rate of 0.3 g supplement/(100 kg body weight day). Serum samples were taken weekly for osteocalcin, and plasma samples were taken on days 0, 42, and 84 for plasma minerals. On days 0, 42, and 84, subjective and objective lameness exams were performed, and radiographs and synovial fluid samples were taken from reference and osteoarthritic joints. Plasma minerals were similar in both groups and were lower on day 84 than on day 0 (P < 0.05). Si supplementation, fed at the manufacturer’s recommended rate, did not improve lameness or radiographs when compared with controls, and supplemented horses did not show greater collagen degradation and/or synthesis markers in synovial fluid than controls, indicating that cartilage turnover remained unaffected. However, a minimum beneficial threshold and range for Si supplementation standardized to body weight need to be established.     

Measurement of articular cartilage stiffness of the femoropatellar, tarsocrural, and metatarsophalangeal joints in horses and comparison with biochemical data

OBJECTIVE: To determine normal cartilage stiffness values in different weight-bearing and non-weight-bearing areas of 3 different equine joints, and to evaluate the relationship between cartilage stiffness and glycosaminoglycan (GAG) and collagen content. STUDY DESIGN: Compressive stiffness of the articular cartilage was measured in 8 horse cadaver femoropatellar (FP), tarsocrural (TC), and metatarsophalangeal (MT) joints. Gross evaluation, collagen content, GAG content, and histologic appearance were assessed for each measurement location. ANIMALS: Eight equine cadavers (4 intact females, 4 castrated males; 7 Quarter Horse or Quarter Horse type, 1 Arabian; aged 4-12 years, weighing 400-550 kg). METHODS: The articular surfaces of 8 equine cadaver FP, TC, and MT joints were grossly evaluated for signs of articular cartilage pathology. Stiffness at preselected sites (FP joint-6 sites; TC joint-3 sites; MT joint-4 sites) was determined using an arthroscopic indentation instrument. Biochemical composition (collagen, GAG content) and histologic evaluation (modified Mankin score) were assessed for each measurement site. RESULTS: All cartilage from all sites evaluated was determined to be normal based on macroscopic and histologic assessments. No significant correlation between Mankin scores and cartilage stiffness values was observed. Site differences in cartilage stiffness were measured in all 3 joints (P<.001). GAG or collagen content had a significant positive correlation with stiffness values in 6 of 13 sites (P<.05, r>0.622, r2>0.387). CONCLUSION: Relative cartilage stiffness values measured in healthy equine joints are site dependent and can be measured using an indentation device intended for arthroscopic application. CLINICAL RELEVANCE: An indentation instrument provided an objective means of determining relative compressive stiffness of articular cartilage. Further research needs to be performed to confirm the site and joint differences observed in this study in clinically normal horses and to determine if the tester can be used clinically to predict articular cartilage pathology.     

The hemodynamic, tissue oxygenation, and selected biochemical effects of isoflurane and halothane anesthesia in horses

Eight healthy horses (337 to 643 kgs) were used to determine the effects of halothane and isoflurane (1.5 MAC) on cardiovascular and blood gas parameters over a 4 hour perio d, while in left lateral recumbency. Each animal was used twice (2 weeks apart) and the order in which the agents were used was randomized. End tidal CO2 was maintained at 45-55 mmHg using positive pressure ventilation. The following parameters were measured every 30 minutes; HR, ET CO2, ET anes. gas cone., SAP, CVP, PAP, and CO. Heparinized samples were also drawn for blood gas analysis from the right lateral metatarsal artery, right jugular vein, and the femoral veins in each rear limb. The 02 delivered to the tissues (TOD) was calculated using the following equation: {ie396-1} It was determined that more 02 was delivered to the tissues during isoflurane anesthesia (382m102/min/m2 + 21 SEM) than halothane anesthesia (280m102min/m2 + 11 SEM). Furthermore the 02 extraction ratio (OER) in the dependent limb was significantly higher during halothane anesthesia. The differences seen in the OER's are due to both anesthetic and positional effects. Serum biochemical changes also suggest differences in tissue damage related to differences in TOD and OER.     

Postpartum supplemental fat, but not maternal body condition score at parturition, affects plasma and adipose tissue fatty acid profiles of suckling beef calves

Three-year-old Angus x Gelbvieh beef cows, which were nutritionally managed to achieve a BCS of 4 +/- 0.07 (479 +/- 36 kg of BW) or 6 +/- 0.07 (580 +/- 53 kg of BW) at parturition, were used in a 2-yr experiment (n = 36/yr) to determine the effects of maternal BCS at parturition and postpartum lipid supplementation on fatty acid profile of suckling calf plasma and adipose tissue. Beginning 3 d postpartum, cows within each BCS were assigned randomly to 1 of 3 treatments in which cows were all fed hay and either a low-fat (control) supplement or supplements with either high-linoleate cracked safflower seeds (linoleate) or high-oleate cracked safflower seeds (oleate) until d 61 of lactation. Diets were formulated to be isonitrogenous and isocaloric, and safflower seed supplements were provided to achieve 5% of DMI as fat. Total concentration of fatty acids in plasma did not differ (P = 0.48) due to maternal BCS at parturition. Percentage of 20:5n-3 in plasma tended (P = 0.06) to be greater for calves suckling cows with a BCS of 6 at parturition. No other differences (P = 0.12 to 0.99) were noted in calf plasma fatty acid profile due to maternal BCS at parturition. Likewise, no differences were detected for total fatty acid concentration (P = 0.88) in calf adipose tissue due to maternal BCS at parturition. Weight percentage of 14:1 (P = 0.001) was greatest in adipose tissue of calves suckling cows fed control and oleate; however, the percentages of 14:0, 15:0, 16:0, 16:1, 17:0, and 18:3n-3 were greater (P < 0.001) in adipose tissue from calves suckling cows fed control compared with calves suckling cows fed linoleate or oleate. Percentages of 18:0, 18:1trans-11, 18:2n-6, and cis-9, trans-11 CLA were greater (P < 0.001) in adipose tissue from calves suckling cows fed linoleate compared with calves suckling cows fed control and oleate. Calves suckling cows fed oleate had greater (P < 0.001) percentages of 18:1trans-9, 18:1trans-10, and 18:1cis-9 in adipose tissue than calves suckling cows fed control or linoleate. Calf plasma and adipose tissue fatty acid profiles were reflective of milk fatty acids. Because fatty acids play an important role in metabolic regulatory functions, changes in milk fatty acid profile should be considered when beef cows are fed lipid supplements.     

Neutrophil myeloperoxidase measurements in plasma, laminar tissue, and skin of horses given black walnut extract

OBJECTIVE: To compare measurements of myeloperoxidase (MPO) in plasma, laminar tissues, and skin obtained from control horses and horses given black walnut heartwood extract (BWHE). ANIMALS: 22 healthy 5- to 15-year-old horses. PROCEDURES: Horses were randomly assigned to 4 groups as follows: a control group given water (n = 5) and 3 experimental groups given BWHE (17) via nasogastric intubation. Experimental groups consisted of 5, 6, and 6 horses that received BWHE and were euthanatized at 1.5, 3, and 12 hours after intubation, respectively. Control horses were euthanatized at 12 hours after intubation. Plasma samples were obtained hourly for all horses. Laminar tissue and skin from the middle region of the neck were harvested at the time of euthanasia. Plasma and tissue MPO concentrations were determined via an ELISA; tissue MPO activity was measured by use of specific immunologic extraction followed by enzymatic detection. RESULTS: Tissues and plasma of horses receiving BWHE contained significantly higher concentrations of MPO beginning at hour 3. Laminar tissue and skin from horses in experimental groups contained significantly higher MPO activity than tissues from control horses. Concentrations and activities of MPO in skin and laminar tissues were similar over time. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, BWHE administration causes increases in MPO concentration and activity in laminar tissue and skin and the time of increased MPO concentration correlates with emigration of WBCs from the vasculature. These findings support the hypothesis that activation of peripheral WBCs is an early step in the pathogenesis of acute laminitis.     

Growth, nutrient utilization, radiographic bone characteristics and postprandial thyroid hormone concentrations in weanling horses fed added dietary fat

Thirty, 19-week-old Quarter Horses were utilized in a 112-d study to compare growth, nutrient utilization and post prandial thyroid hormone concentrations in weanling horses fed fat-supplemented or conventional diets. Concentrates were formulated to contain the same nutrient-calorie ratios and were fed with Coastal Bermuda grass hay in a 70:30 concentrate:hay ratio. Initially and at 28-d intervals the horses were weighed and measured for height, heart girth circumfer ence and subcutaneous fat thickness over the rump and ribs. Radiographs were taken of the right carpal and metacarpal phalangealjoints at the beginning and end of the experimental period for evaluation of bone density and physeal maturation. A digestion trial was conducted on each horse 10 d prior to completion of the study. On d 70, blood samples were col lected from 6 females in each treatment group over a 6-h collection period for evaluation of postprandial insulin, T₄ and T₃ concentrations in response to the diets. Horses in both groups consumed an average of 2.6% of body weight daily of total feed (Dry Matter). Weanlings consuming the fat-supplemented concentrate tended to have higher (P = .10) average daily weight gains (.80 vs .74 kg) and lower (P<.05) feed:gain ratios (6.3 vs 7.3) than weanlings fed the control diet. Gain in heart girth circumference was greater (P<.05) for the fat fed weanlings (20.1 cm) than the control weanlings (17.9 cm). All other linear measurements of growth were similar between treatments. There were no radiographic indications of abnormalities in the physes of horses on either treatment. Closure of the physes occurred at normal rates in foals on both treatments. Radiographic bone density was not different between treatments, and there was an expected increase in bone density in weanlings on both treatments over time. Digestibilities of protein and energy were similar between treatments, while ether extract and neutral detergent fiber digestibilities were higher (P<.05 and P<.06, respec tively) in those horses consuming the fat-supplemented diet (65.39 vs 57.67% and40.55 vs 35.62%, respectively). These data indicate that fat can be used as an energy source to support growth and development in weanling horses.     

Anti-Trichinella antibodies detected in chronically infected horses by IFA and Western blot, but not by ELISA

In the Balkan countries, where trichinellosis is a re-emerging zoonosis, it is of great importance to determine Trichinella infection prevalence among the major hosts, including horses. One method for monitoring prevalence is serological surveillance; however, the validity of serological methods in horses is not well understood. The dynamics of anti-Trichinella IgG production and circulating excretory/secretory (ES) antigens were investigated in three horses experimentally-infected with Trichinella spiralis. Horses were slaughtered at 32 week post infection (p.i.). Low worm burdens were found in all three animals. Anti-Trichinella IgG was detected up to 32 weeks p.i. by an indirect immunofluorescence assay (IFA) and by Western blot (Wb), but not by ELISA. The ELISA test detected antibodies for only a short period of time (up to 18 weeks p.i. using ES antigen or up to 20 weeks p.i. using tyvelose-BSA antigen). The presence of circulating muscle larvae ES antigen in sera of infected horses was observed by dot blot from the 4th week p.i. up to the 32nd week p.i.     

Biochemical characterisation of navicular hyaline cartilage,navicular fibrocartilage and the deep digital flexor tendon in horses with navicular disease

The study hypothesis was that navicular disease is a process analogous to degenerative joint disease, which leads to changes in navicular fibrocartilage and in deep digital flexor tendon (DDFT) matrix composition and that the process extends to the adjacent distal interphalangeal joint. The objectives were to compare the biochemical composition of the navicular articular and palmar cartilages from 18 horses with navicular disease with 49 horses with no history of front limb lameness, and to compare navicular fibrocartilage with medial meniscus of the stifle and collateral cartilage of the hoof. Cartilage oligomeric matrix protein (COMP), deoxyribonucleic acid (DNA), total glycosaminoglycan (GAG), metalloproteinases MMP-2 and MMP-9 and water content in tissues were measured. Hyaline cartilage had the highest content of COMP and COMP content in hyaline cartilage and tendon was higher in lame horses than in sound horses (p<0.05). The concentration of MMP-2 amount in hyaline cartilage was higher in lame horses than in sound horses. The MMP-2 amounts were significantly higher in tendons compared to other tissue types. Overall, 79% of the lame horses with lesions had MMP-9 in their tendons and the amount was higher than in sound horses (p<0.05). In horses with navicular disease there were matrix changes in navicular hyaline and fibrocartilage as well as the DDFT with potential implications for the pathogenesis and management of the condition.     

Age-related changes in serum insulin-like growth factor-I, insulin-like growth factor-I binding protein-3 and articular cartilage structure in Thoroughbred horses

REASONS FOR PERFORMING STUDY: Structural changes in articular cartilage associated with the ageing process require definition for investigators performing developmental and age-related studies, for which information is lacking. OBJECTIVES: To 1) determine the onset and end of puberty as defined by serum insulin like growth factor (IGF-I) and IGF-binding protein-3 (IGFBP-3) concentrations and 2) correlate articular-epiphyseal cartilage complex structural changes with the onset and end of puberty. METHODS: IGF-I and IGFBP-3 were measured in serum samples from normal female and male horses age 9-715 days to determine peak and steady-state values for horses transitioning through puberty. Osteochondral tissue sections were obtained from horses age 120-840 days (4-28 months) and examined histologically for cartilage canals and tidemark formation. RESULTS: In male and female horses, serum IGF-I/IGFBP-3 concentrations peaked at approximately 225 days, defining the onset of puberty. Cartilage canals were absent from articular cartilage just prior to this time point. IGF-I/IGFBP-3 concentrations declined to steady-state levels at approximately age 450 days, signalling exit from puberty and therefore the beginning of ageing. This time point correlated to initial formation of a tidemark in the osteochondral tissue sections. CONCLUSIONS: Horses may be considered pubescent at age 225-450 days, and post pubescent and ageing after age 450 days. POTENTIAL RELEVANCE: Defining the normal post natal to post pubescent concentrations for serum IGF-I and serum IGFBP-3 establishes subsets of animals for age-related studies and may be used to monitor horses for abnormally high IGF-I concentrations due to natural disease or subsequent to systemic growth hormone administration.     

Behavior of back fat thickness, the activity of NADP-dependent dehydrogenases from adipose tissue and adipose tissue constituents fat and protein and their evidence for energy metabolism in dairy cows

Studies were conducted into 60 Friesian dairy cattle in the GDR for determination of dorsal fat thickness (DFT), activities of glucose-6-phosphate dehydrogenase (GPDH) and isocitrate dehydrogenase (ICDH) in adipose tissue, concentrations of fat and protein in adipose tissue, 2 weeks ante partum as well as 0, 2, 4, 6, 8, 12, 16, 20, 28, 36 weeks post partum, and liver fat levels, 2 and 4 weeks post partum. DFT, ICDH, GPDH, ICDH-GPDH ratio, fat level, fat-protein quotient, changes in DFT, GPDH, and fat-protein quotient exhibited significant relations with the weeks of lactation. The above 60 experimental cows were subdivided by 6 groups of half-siblings consisting of 10 animals each. Significant differences were found to exist between these groups of half-siblings with regard to DFT, GPDH, ICDH-GPDH ratio, and fat-protein quotient. Within each of the half-sibling groups, significant differences were found to exist between individuals for DFT, ICDH, GPDH, ICDH-GPDH ratio, and fat-protein quotient. The above parameters can be used to describe the energy metabolism of dairy cow via quantitative and temporal curves of fat mobilisation and fat deposition. In the context of both animal health and breeding, more attention should be given to determination of mobilisation and deposition of fat as well as to the post partum energy deficit.     

Wet litter not only induces footpad dermatitis but also reduces overall welfare,technical performance,and carcass yield in broiler chickens

This study investigated whether a high level of footpad dermatitis (FPD) in broiler chickens induced by increased litter moisture content is accompanied by negative effects on technical performance, carcass yield, and other welfare aspects. Litter moisture content was increased by systematically spraying water over the litter from 6 d of age onward (lesion-induction treatment). Results were compared with a control group kept on relatively good quality litter and having a very low prevalence of footpad lesions. Litter quality significantly decreased from 7 d of age onward in the pens with the lesion-induction treatment compared with the control pens. At 21 and 36 d of age, significantly more FPD was observed in the lesion-induction groups compared with the control groups. Technical performance of the broilers was negatively affected for the lesion-induction groups from 28 d of age onward; lesion-induction groups had significantly lower BW gain, feed intake, and water intake and significantly higher FCR compared with control groups. As a result of the lower BW gain, carcass weight was less for the lesion-induction group and significantly more rejections for commercial parts were found. The lesion-induction groups also had significantly more hock burns and breast irritations at d 21 and 36, and were dirtier but had fewer thigh scratches at d 35 compared with the control groups. In addition, locomotion was negatively affected among the lesion-induction group, as they had a higher gait score at d 36 compared with the control group. We concluded that increased litter moisture content not only caused severe FPD but also reduced broiler performance and carcass yield and had a negative effect on other welfare aspects.     

The effect of dietary energy source on serum concentration of insulin-like growth factor-I growth hormone,insulin, glucose,and fat metabolites in weanling horses

Feeding diets high in soluble carbohydrates to growing horses has been implicated in the development of orthopedic diseases; as a result, substitution of dietary fat for soluble carbohydrates has received attention. Because IGF-I is integral to growth and cartilage development and because it is influenced by nutrition, we evaluated the effect of dietary fat substitution on metabolic endpoints and circulating GH and IGF-I in growing horses. Twelve Quarter Horse weanlings, four female and eight male, 151 to 226 d old, were blocked by sex and age and assigned to two treatment groups. Group one (CARB; n = six) was fed a concentrate containing 2.21% fat and 33.9% starch; group two (FAT; n = six) was fed a concentrate containing 10.3% fat and 24.0% starch. Both concentrates contained 3.0 Mcal/kg of DE and 16% CP. Brome hay also was fed. Diets were fed at 0800 and 1600 for 60 d. On d 0, 30, and 60, blood samples were obtained via a jugular catheter from 1 h before until 5 h after the morning feeding. Serum was analyzed for glucose, insulin, GH, IGF-I, NEFA, and total cholesterol (CHOL). Neither ADG (0.85 +/- 0.04 and 0.84 +/- 0.04 kg) nor concentrate DMI (4.04 +/- 0.12 and 4.03 +/- 0.12 kg/d) differed between treatments. There were consistent increases in glucose and insulin in response to feeding on d 0, 30, and 60 for both groups. On d 30, the glucose response to feeding was less (P = 0.07) over time in FAT vs. CARB; however, there were no significant treatment x time effects on d 0 or 60. On d 60, the insulin response to feeding was less (P < 0.05) over time in FAT compared with CARB; however, there was no treatment x time effect on d 0 or 30. Serum CHOL concentrations did not differ between groups on d 0. Horses in the FAT group had increased CHOL concentrations on d 30 and 60 compared with CARB (P < 0.01). Although treatment x time interactions were noted for GH on d 30 and 60 (P < 0.05), only transient and inconsistent differences in the secretory profiles between CARB and FAT treatments were evident at those sampling times. Serum NEFA and IGF-I did not differ between treatments on d 0, 30, or 60. These results suggest that dietary energy source, at least at the level used in this study, did not affect foal growth performance or serum IGF-I and NEFA concentrations. Fat substitution increased serum CHOL and variably affected serum GH, glucose, and insulin concentrations in response to feeding.     

日粮中添加菊粉和不同脂肪类型对肉鸡行为、血清代谢物、肝脏脂肪、腹脂沉积以及组织中脂肪酸成分的影响

该试验旨在评价2种不同脂肪来源的饲料中添加菊粉对肉鸡行为、血清代谢物、肝脏脂质、腹脂、胸肌和腿肌中脂肪酸组成的影响。将240只1日龄雌性肉鸡随机分成6组,每组8个重复,每个重复5只肉鸡。试验采用3×2双因子试验设计,菊粉设3个水平（0、5和10g/kg）,日粮中2种类型的脂肪[棕榈油（PO）组和葵花籽油（SO）组]含量均为90g/kg。试验期为1～34d。日粮中的脂肪不影响肉鸡的体重增长,但PO组肉鸡的消化率低于SO组（p〈0.001）。日粮中含棕榈油增加了肉鸡腹脂沉积和血清中脂质及葡萄糖的含量。PO组肉鸡肝脏中的三酰甘油含量高于SO组。同样,日粮中的脂肪改变了肉鸡腹脂和肌内的脂肪酸组成,导致PO组肉鸡的C16：0和C18：1n-9含量高于SO组,C18：2n-6含量却更低。日粮中添加菊粉对肉鸡体重的增长（p=0.017）有二次效应,而对消化率没有影响。与对照组相比,菊粉减少了肉鸡肝脏油脂的总含量（p=0.003）、血清中甘油三酯含量以及低密度的脂蛋白胆固醇（31%）。菊粉添加到含葵花籽油的日粮中,增加了肉鸡腹脂和肌内多不饱和脂肪酸与饱和脂肪酸的比值。该研究结果表明：日粮中添加菊粉对肉鸡血脂的有益作用,是因为减少了三酰甘油含量。该结果同样证明,菊粉增强了葵花籽油提高肉鸡肌内多不饱和脂肪酸与饱和脂肪酸比值的能力。