•Maternal overnutrition shifts muscle fiber type differentiation in the foal.•Skeletal muscle fiber area is increased in foals from overfed mares.•Foal gene expression and cell signaling were altered ...by maternal overnutrition.
Skeletal muscle plays an integral role in the ability of a horse to perform at high levels. Shifts in skeletal muscle development in response to maternal plane of nutrition may have substantial and lasting impacts on athletic performance and whole-body metabolism. Therefore, sixteen Quarter Horse mares were used in a completely randomized design and maintained at a body condition score (BCS) 6 until start of third trimester. On d 235 of gestation, mares were randomly assigned to receive one of two dietary treatments with a diet formulated to meet requirements during late gestation (CON; n = 8), and an overfed diet (HIGH; n = 8) where mares received an additional 40% above CON. Five h after parturition, foals were euthanized, and gluteus medius, triceps brachii, and semitendinosus were harvested for analyses. Gene expression was determined by qPCR and western immunoblotting was used to quantify total and phosphorylated forms of proteins involved in skeletal muscle metabolism with tubulin as the loading control. All data were analyzed using PROC MIXED of SAS. Foals from HIGH mares exhibited larger skeletal muscle fibers by area (P <0.05), and a shift in muscle fiber development towards type I slow twitch muscle fibers (P <0.05). Relative expression of glucose transporter 4 (GLUT4) was lower in HIGH foals compared to CON in gluteus medius (P = 0.05). Insulin receptor isoform B (INSR-B) and insulin-like growth factor 1 receptor (IGF1R) were greater in triceps brachii of HIGH foals compared to CON (P ≤ 0.03). Insulin receptor isoform A (INSR-A), however, tended to be lower in triceps brachii of HIGH compared to CON (P = 0.10). Ratios of phosphorylated to total extracellular signal-regulated protein kinase 1/2 (ERK1/2) and c-June N-terminal kinase (JNK) were higher in HIGH foals compared to CON (P ≤0.04) in gluteus medius. There were no differences observed for phosphorylated to total protein ratios in semitendinosus and triceps brachii muscles; however, total ERK1/2 tended to be elevated (P <0.10) in semitendinosus from CON foals compared to HIGH. There was no difference in phosphorylated or total protein kinase B (AKT) (P >0.14). These data indicate hypertrophy of skeletal muscle fibers and a shift towards type I slow twitch fibers in HIGH foals. Furthermore, this study identifies muscle specific changes in gene expression and downstream insulin receptor signaling, which may contribute to future metabolic abnormalities in response to maternal overnutrition.
•Overnutrition during late gestation alters maternal insulin sensitivity.•Offspring pancreas size not influenced by maternal overnutrition.•Spleen and testes larger in foals of overfed ...mares.•Increased size and abundance of pancreatic islets of Langerhans in foals from overfed mares.
Results from previous studies indicate that maternal overnutrition during late gestation predisposes foals to metabolic disease, however, specific mechanisms resulting in disease remain unknown. Quarter Horse mares (n = 16), were randomly assigned to dietary treatments, beginning on gestational day 235, and consisted of a control group (CON- diet meeting nutrient requirement; n = 8) or an overfed diet (HIGH; n = 8) where mares received an additional 40 % above CON. On gestational days 285 and 315, an intravenous glucose tolerance test (FSIGTT) was conducted. Following parturition, foals were separated from the mare, prohibited from nursing, and an FSIGTT was conducted at 2 h postpartum. Foals were immediately euthanized and tissues preserved for analyses. There was no effect of treatment on foal BW (P = 0.50), pancreas weight (P = 0.60), or FSIGTT area under the curve for glucose (P = 0.80) and insulin (P = 0.70). Colocalization of α-amylase to isolate pancreatic islets of Langerhans indicated increased islet number and size in foals from HIGH mares (P < 0.01). Immunofluoresent analysis of insulin, glucagon, and somatostatin indicate no difference in intensity of staining (P> 0.10). Foals exposed to overnutrition during peak fetal growth had altered pancreatic islet development that may lead to adult-onset metabolic disease.
Eighteen Quarter Horses were used in a randomized complete design for a 28-d experiment to evaluate age-related effects on inflammation and cartilage turnover after induction of a single inflammatory ...insult using lipopolysaccharide (LPS). Horses were grouped by age as yearlings (3 males and 3 females), 2 to 3 yr olds (2/3 yr old; 2 males and 4 females), and skeletally mature 5 to 8 yr olds (mature; 2 males and 4 females). On d 0, all horses were individually housed and fed diets that met or exceeded requirements. On d 14, horses were challenged with an intra-articular injection of LPS. Radial carpal joints were randomly assigned to receive 0.5 ng LPS solution obtained from O55:B5 or 0.8 mL sterile lactated Ringer's solution as a contralateral control. Synovial fluid was collected prior to LPS injection at h 0 before injection and at 6, 12, 24, 168, and 336 h after injection. Samples were analyzed using commercial ELISA kits for PGE, collagenase cleavage neoepitope (C2C), and carboxypropeptide of type II collagen (CPII). Heart rate (HR), respiratory rate (RR), and rectal temperature (RT) were monitored over the initial 24 h and carpal circumference and surface temperature were also recorded, with additional measurements at 168 and 336 h. Data were analyzed using PROC MIXED of SAS. Values for RT, HR, and RR were within the normal range for each age group. Heart rate and RT were influenced by age ( < 0.01), whereas RR was unaffected ( ≤ 0.21). Joint circumference was not influenced by age of horse ( = 0.84), but circumference and surface temperature increased ( < 0.01) over time across all age groups. Synovial PGE concentrations tended ( = 0.09) to be influenced by age, with yearlings having lower ( = 0.03) concentrations than mature horses. Concentrations of synovial C2C were affected by age of horse, with yearlings and 2/3 yr olds having lower ( < 0.01) concentrations than mature horses. Similarly, synovial CPII was influenced by age, with yearlings and 2/3 yr olds having lower ( ≤ 0.02) concentrations than mature horses. Ratios of anabolic CPII to catabolic C2C varied by age, with mature and 2/3-yr-old horses having greater ( < 0.01) values compared with yearlings. These results indicate that inflammation and the corresponding cartilage turnover in response to LPS administration vary with age.
Abstract
Twenty-seven mature Quarter horses were used in a randomized design to determine the effects of bioactive protein supplementation on gait kinematics and systemic inflammatory markers in a ...34-d trial. Treatments consisted of oral doses of 230 g/d of pelleted supplements containing 0 g (CON; n = 9), 40 g of bioactive protein (40BP; n = 9; LIFELINE, APC, LLC, Ankeny, IA), and 80 g of bioactive protein (80BP; n = 9) daily. Horses were fed a commercial concentrate at 0.5% BW (as-fed) and received ad libitum coastal bermudagrass (Cynodon dactylon) hay daily. On day 33, horses consistent in exercise (CON, n = 6; 40BP, n = 8; 80BP, n = 7) participated in a trailering and riding challenge. Kinematic gait analysis was performed on day 0 for use as a covariate, and on day 14, 28, and 34 to allow for the determination of potential time and dosage effects. Video footage was collected and analyzed using gait analysis software (EquineTec, Monroe, GA) for the determination of stride length (SL) and range of motion (ROM). Blood was collected via jugular venipuncture on days 0, 14, 28, and 34 for determination of systemic expression of tumor necrosis factor (TNF)-α and IL-1β. Data were analyzed using PROC MIXED of SAS. A trend towards treatment × time interaction was observed in ROM of the knee at the walk (P = 0.10), due to the increasing ROM for 40BP and 80BP as time increased and decreasing ROM for CON. A treatment × time interaction was observed (P < 0.01) for hock ROM at a walk resulting from CON and 80BP decreasing from day 14 to 28 with 40BP increasing, while from day 28 to 34 ROM at a walk decreased for 40BP and increased for 80BP. The main effect of treatment on hock ROM at the walk was quadratic (P < 0.01) and characterized by higher ROM values for 40BP compared to CON or 80BP. Dietary treatment lengthened (P = 0.04) SL of the hind limb at the walk for 40BP and 80BP compared to CON on both days 14 and 28. A significant treatment × time interaction was observed in the expression of IL-1β (P < 0.01) and can be explained by lower concentrations of IL-1β for 80BP on day 34 compared to the other treatments, with 40BP being intermediate and CON being the highest. Increased articular ROM with decreased expression of IL-1β may indicate potential anti-inflammatory effects of 80 g/d of bioactive proteins.
Eighteen Quarter Horses were used in a randomized complete design for a 28-d experiment to evaluate age-related effects on inflammation and cartilage turnover after induction of a single inflammatory ...insult using lipopolysaccharide (LPS). Horses were grouped by age as yearlings (3 males and 3 females), 2 to 3 yr olds (2/3 yr old; 2 males and 4 females), and skeletally mature 5 to 8 yr olds (mature; 2 males and 4 females). On d 0, all horses were individually housed and fed diets that met or exceeded requirements. On d 14, horses were challenged with an intra-articular injection of LPS. Radial carpal joints were randomly assigned to receive 0.5 ng LPS solution obtained from O55:B5 or 0.8 mL sterile lactated Ringer's solution as a contralateral control. Synovial fluid was collected prior to LPS injection at h 0 before injection and at 6, 12, 24, 168, and 336 h after injection. Samples were analyzed using commercial ELISA kits for PGE, collagenase cleavage neoepitope (C2C), and carboxypropeptide of type II collagen (CPII). Heart rate (HR), respiratory rate (RR), and rectal temperature (RT) were monitored over the initial 24 h and carpal circumference and surface temperature were also recorded, with additional measurements at 168 and 336 h. Data were analyzed using PROC MIXED of SAS. Values for RT, HR, and RR were within the normal range for each age group. Heart rate and RT were influenced by age ( < 0.01), whereas RR was unaffected ( ≤ 0.21). Joint circumference was not influenced by age of horse ( = 0.84), but circumference and surface temperature increased ( < 0.01) over time across all age groups. Synovial PGE concentrations tended ( = 0.09) to be influenced by age, with yearlings having lower ( = 0.03) concentrations than mature horses. Concentrations of synovial C2C were affected by age of horse, with yearlings and 2/3 yr olds having lower ( < 0.01) concentrations than mature horses. Similarly, synovial CPII was influenced by age, with yearlings and 2/3 yr olds having lower ( ≤ 0.02) concentrations than mature horses. Ratios of anabolic CPII to catabolic C2C varied by age, with mature and 2/3-yr-old horses having greater ( < 0.01) values compared with yearlings. These results indicate that inflammation and the corresponding cartilage turnover in response to LPS administration vary with age.
Bisphosphonates are commonly used to treat and prevent bone loss, but their effects in active, juvenile populations are unknown. This study examined the effects of intramuscular clodronate disodium ...(CLO) on bone turnover, serum bone biomarkers (SBB), bone mineral density (BMD), bone microstructure, biomechanical testing (BT), and cartilage glycosaminoglycan content (GAG) over 165 days. Forty juvenile sheep (253 ± 6 days of age) were divided into four groups: Control (saline), T0 (0.6 mg/kg CLO on day 0), T84 (0.6 mg/kg CLO on day 84), and T0+84 (0.6 mg/kg CLO on days 0 and 84). Sheep were exercised 4 days/week and underwent physical and lameness examinations every 14 days. Blood samples were collected for SBB every 28 days. Microstructure and BMD were calculated from tuber coxae (TC) biopsies (days 84 and 165) and bone healing was assessed by examining the prior biopsy site. BT and GAG were evaluated postmortem. Data, except lameness data, were analyzed using a mixed-effects model; lameness data were analyzed as ordinal data using a cumulative logistic model. CLO did not have any measurable effects on the skeleton of sheep. SBB showed changes over time (p ≤ 0.03), with increases in bone formation and decreases in some bone resorption markers. TC biopsies showed increasing bone volume fraction, trabecular spacing and thickness, and reduced trabecular number on day 165 versus day 84 (p ≤ 0.04). These changes may be attributed to exercise or growth. The absence of a treatment effect may be explained by the lower CLO dose used in large animals compared to humans. Further research is needed to examine whether low doses of bisphosphonates may be used in active juvenile populations for analgesia without evidence of bone changes.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Abstract
Sixteen weanling Quarter horses (255 ± 22 kg BW) were used in a 56-d trial to determine effects of Zn, Cu, Mn, and Co source on intra-articular inflammation and cartilage metabolism ...following an inflammatory insult. Horses were stratified by days of age (233 ± 20 d), sex (7 fillies, 9 colts), horse source (7 TAMU, 9 Birdsong Farms), and BW. Horses were randomly assigned to dietary treatment formulated with inorganic sources (CON; CuSO4, ZnSO4, and MnSO4 and CoCO3; n=8) or metal complexes (ZPM; zinc methionine, manganese methionine, copper lysine, cobalt glucoheptonate; n=8). Added trace minerals were at iso-levels between treatments and trace mineral intakes met or exceeded NRC requirements. Horses were offered 1.75% BW treatments (as-fed) and 0.75% BW coastal bermudagrass (Cynodon dactylon; as-fed) hay. Total diet was split into two equal feedings daily at 0600 and 1600h. Daily DM intakes did not differ between treatments (P>0.05). On d 42, an intra-articular lipopolysaccharide (LPS) challenge was conducted. Carpal joints were randomly assigned to receive intra-articular injections of 0.5 ng LPS derived from Escherichia coli 055:B5 or sterile lactated Ringer’s solution as a contralateral control. Synovial fluid was collected at pre-injection h0 and 6, 12, 24, 168, and 336h post-injection. Samples were analyzed for prostaglandin E2 (PGE2), carboxypeptide of type II collagen (CPII), collagenase cleavage neopeptide (C2C) and aggrecan chondroitin sulfate 846 epitope (CS846) using commercial ELISA kits. Data were analyzed using the MIXED procedure of SAS. Intra-articular LPS increased (P≤0.01) synovial PGE2, C2C, CPII, and CS846 concentrations regardless of treatment. Synovial C2C was greater (P<0.01) in ZPM horses (373.5 ± 9.3 ng/mL) compared to CON (337.4 ± 9.3 ng/mL). In contrast, CPII concentration remained unaffected by treatment. Despite differences in degradation, the ratio of anabolic to catabolic processes (CPII:C2C) was unaffected by diet (P=0.57). No effect of diet was observed for PGE2 (P=0.13); however, ZPM horses had a greater increase in concentration of PGE2 in the LPS-injected joint at 6h compared to CON (P<0.01; 3239.7 ± 116.2 and 2664.8 ± 116.2 pg/mL, respectively). A diet time LPS interaction was observed for CS846. Concentrations for ZPM horses peaked at 12h while CON horses peaked with a lower concentration at 24h (P<0.05). Compared to inorganic mineral sources, data suggest that sufficient intake of a complexed trace mineral source may support a more robust proinflammatory immune response to an LPS challenge resulting in increased collagen degradation and a more rapid rise in aggrecan reparative processes in young growing horses.
Sixteen weanling Quarter horses (255 ± 22 kg BW) were used in a 56-d trial to determine effects of Zn, Cu, Mn, and Co source on intra-articular inflammation and cartilage metabolism following an ...inflammatory insult. Horses were stratified by days of age (233 ± 20 d), sex (7 fillies, 9 colts), horse source (7 TAMU, 9 Birdsong Farms), and BW. Horses were randomly assigned to dietary treatment formulated with inorganic sources (CON; CuSO4, ZnSO4, and MnSO4 and C°CO3; n=8) or metal complexes (ZPM; zinc methionine, manganese methionine, copper lysine, cobalt glucoheptonate; n=8). Added trace minerals were at iso-levels between treatments and trace mineral intakes met or exceeded NRC requirements. Horses were offered 1.75% BW treatments (as-fed) and 0.75% BW coastal bermudagrass (Cynodon dactylon; as-fed) hay. Total diet was split into two equal feedings daily at 0600 and 1600h. Daily DM intakes did not differ between treatments (P>0.05). On d 42, an intra-articular lipopolysaccharide (LPS) challenge was conducted. Carpal joints were randomly assigned to receive intra-articular injections of 0.5 ng LPS derived from Escherichia coli 055:B5 or sterile lactated Ringer's solution as a contralateral control. Synovial fluid was collected at pre-injection h0 and 6, 12, 24, 168, and 336h post-injection. Samples were analyzed for prostaglandin E2 (PGE2), carboxypeptide of type II collagen (CPII), collagenase cleavage neopeptide (C2C) and aggrecan chondroitin sulfate 846 epitope (CS846) using commercial ELISA kits. Data were analyzed using the MIXED procedure of SAS. Intra-articular LPS increased (P=0.01) synovial PGE2, C2C, CPII, and CS846 concentrations regardless of treatment. Synovial C2C was greater (P<0.01) in ZPM horses (373.5 ± 9.3 ng/mL) compared to CON (337.4 ± 9.3 ng/mL). In contrast, CPII concentration remained unaffected by treatment. Despite differences in degradation, the ratio of anabolic to catabolic processes (CPII:C2C) was unaffected by diet (P=0.57). No effect of diet was observed for PGE2 (P=0.13); however, ZPM horses had a greater increase in concentration of PGE2 in the LPS-injected joint at 6h compared to CON (P<0.01; 3239.7 ± 116.2 and 2664.8 ± 116.2 pg/mL, respectively). A diet time LPS interaction was observed for CS846. Concentrations for ZPM horses peaked at 12h while CON horses peaked with a lower concentration at 24h (P<0.05). Compared to inorganic mineral sources, data suggest that sufficient intake of a complexed trace mineral source may support a more robust proinflammatory immune response to an LPS challenge resulting in increased collagen degradation and a more rapid rise in aggrecan reparative processes in young growing horses.
Abstract This experiment evaluated the effects of multiple bovine-appeasing substance (BAS) administration during a 42-d preconditioning program followed by a feedlot receiving period on ...productivity, health, and physiological variables of feeder cattle. Ninety calves were weaned, weighed, loaded into a livestock trailer, transported for 70 km, and unloaded at the Bozeman Agricultural Research and Teaching Farm for a 42-d preconditioning program. Upon arrival, calf body weight (BW) was recorded again, and both pre- and post-transport BWs were averaged and used as calf weaning initial BW. Calves were ranked by BW, sex, and age in a completely randomized design and assigned to receive 1) multiple administrations of BAS at weaning (day 0), days 14, 28, and before transport and feedlot entry (day 42; BAS; RSEA Group, Quartier Salignan, France; n = 9 pens/treatment), or 2) placebo (diethylene glycol monoethyl ether; CON; n = 9 pens/treatment). Treatments (5 mL) were applied to the nuchal skin area of each animal during the preconditioning period. Calves within treatment groups were ranked again by initial BW, sex, and age, in a manner that pens have similar initial BW, age, and three steers and two heifers and allocated to 1 of the18 drylot pens. On day 42, calves were combined within the treatment group, loaded into two different single double-deck commercial livestock trailers, and transported for 1,000 km (approximately 16 h). Upon arrival (day 43), calves were unloaded at the same feedyard. Blood samples were collected on days 0, 3, 7, 14, 21, 28, 42, 43, 46, 50, 57, 64, and 90. Average daily gain, final BW, and feed efficiency did not differ (P > 0.52) between BAS and CON calves in the preconditioning and receiving phases. A treatment × day interaction was detected (P < 0.001) for plasma haptoglobin concentrations, which was greater (P < 0.01) in CON on days 3 and 7 vs. BAS calves. During the preconditioning phase, serum NEFA concentration was reduced (P < 0.01) in BAS on day 3 compared with CON calves. A treatment × day interaction was detected (P = 0.001) for exit velocity, which was greater (P < 0.001) for CON vs. BAS calves on days 3, 7, 14, and 21 during the preconditioning phase and on day 46 of the receiving phase. Therefore, Applications of BAS reduced immunological responses and exit velocity associated with stress caused by management practices, but did not improve performance during the preconditioning and receiving phases.
Abstract
Passive immune transfer in a newborn foal is highly dependent upon the quality and concentration of immunoglobulins (Ig) in maternal colostrum, primarily IgG. Anti-endotoxin IgY derived from ...chicken egg yolks has been used in calves to reduce morbidity and mortality. Its use may provide additional pathogen-specific immune protection to neonatal foals, but passive transfer has not been confirmed. Therefore, the objective of this study was to quantify passive IgY transfer in neonatal foals given an egg-derived anti-endotoxin IgY supplement (Camas, Inc.) and identify effects on passive immunity. To test this, 5 neonatal foals were randomly divided into two treatment groups. Within 20 minutes after first nursing, the treated group (TRT; n = 3) was administered 40 g of the egg yolk supplement diluted in 50 mL water orally via syringe and the control group (CON; n = 2) was given isovolumetric water. At birth, physical measurements including BW, wither height (WH), hip height (HH), body length (BL), and heart girth (HG) circumference were recorded. Serum samples were collected prior to first nursing (h 0) and h 2, 6, 12, 24, 72, 168, 336 post treatment administration. Rectal temperature, heart rate, and respiration rate were recorded at all blood collection timepoints. Serum samples were analyzed for IgG, IgA, and IgY by commercial ELISA. Custom ELISA was used to identify endotoxin-specific IgY for Salmonella and E. coli at h 6. Western immunoblotting was used to confirm that passively transferred IgY was intact. Data were analyzed using PROC MIXED in SAS. Vital signs remained within normal, healthy ranges and physical measurements at birth were similar for all foals (P > 0.26). There was an effect of treatment (P = 0.05), time (P < 0.01), and treatment × time interaction (P < 0.01) for serum IgY concentrations where IgY was only detectable in TRT foals and peaked at h 6 and 12, decreasing thereafter to h 336. Concentrations of IgG and IgA increased from h 0 to 12 (P < 0.01); however, there were no effects of treatment nor treatment × time interactions (P > 0.24) for either Ig. Western immunoblotting confirmed that IgY was transferred intact and custom ELISA identified IgY specific against Salmonella and Escherichia coli. These results confirm the intact passive transfer of IgY in neonatal foals when given an IgY-infused egg yolk supplement with no negative impact on passive transfer of IgG and IgA from maternal colostrum.
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