Dietary fructan carbohydrate increases amine production in the equine large intestine: implications for pasture-associated laminitis

Pasture-induced laminitis in the horse is associated with the overconsumption of fermentable carbohydrate, in the form of simple sugars, fructans, or starch. The fermentation of carbohydrate in the cecum and large intestine results in the production of lactic acid and other toxins or "laminitis trigger factors." Vasoactive amines have been suggested as possible initiating factors. The aim of this study was to feed a commercially available form of fructan carbohydrate (inulin, 3 g/kg of BW per day) to normal ponies and to ponies predisposed to laminitis, to mimic a change from a basal hay diet to lush spring-summer pasture. Five normal and 6 laminitis-prone, native-breed ponies were acclimated to a basal hay diet before the inclusion of inulin and chopped dried grass. Blood samples, fecal samples, and foot temperature measurements were taken throughout the study. Amines were measured in the feces and plasma by HPLC and liquid chromatography-mass spectrometry, respectively. The pH of the fecal samples decreased from 6.89 +/- 0.11 on the hay diet to a minimum of 6.18 +/- 0.11 with the addition of inulin (P <0.05). An increase was observed in the fecal concentrations of a number of amines, including tryptamine (2.5-fold increase, P <0.05) and tyramine (2-fold increase, P <0.05). No changes were noted in plasma amine concentrations or plasma D- or L-lactate, indicating that there may be a threshold of hindgut pH change before mucosal damage can result in the release of these factors into the circulation. No differences in pH or any of the measured compounds were observed between the group of normal ponies and those predisposed to laminitis. This indicates that differences in the intestinal microflora do not account for this predisposition. However, the results from this study indicate that moderate increases in dietary fructan carbohydrate can produce increases in bacterial fermentation products and other compounds in the large intestine, which may be relevant to the pathogenesis of acute laminitis in ponies on pasture.     

Effect of feeding glucose, fructose, and inulin on blood glucose and insulin concentrations in normal ponies and those predisposed to laminitis

Identification of ponies (Equus caballus) at increased risk of pasture-associated laminitis would aid in the prevention of the disease. Insulin resistance has been associated with laminitis and could be used to identify susceptible individuals. Insulin resistance may be diagnosed by feeding supplementary water-soluble carbohydrate (WSC) and measuring blood glucose and insulin concentrations. The aim of this study was to assess the glycemic and insulinemic responses of 7 normal (NP) and 5 previously laminitic (PLP), mixed breed, native UK ponies fed glucose, fructose, and inulin [1 g/(kg(.)d) for 3 d] or no supplementary WSC (control) in spring and fall after a 7-d adaptation to a pasture or hay diet. Blood samples were taken for 12 h after feeding on each day, and baseline and peak concentrations and area under the curve (AUC) for glucose and insulin were recorded. Linear mixed models were used for statistical analysis. Differences between PLP and NP groups were most marked after glucose feeding with differences in peak glucose (P = 0.02) and peak insulin (P = 0.016) concentrations. Season and diet adaptation also affected results. Peak concentrations of glucose and insulin occurred 2 to 4 h after WSC feeding. Peak insulin concentration was greater and more variable in fall, particularly in PLP adapted to fall pasture. Baseline glucose and insulin concentrations varied between individuals and with season and diet adaptation but were not greater in PLP than NP. Insulin AUC was greater in PLP than NP after feeding both glucose and fructose (P = 0.017), but there were no differences between PLP and NP in glucose AUC. Glycemic and insulinemic changes were less (P ≤ 0.05) after feeding fructose than glucose, although differences between PLP and NP were still evident. Minimal changes in glucose and insulin concentrations occurred after inulin feeding. Measurement of peak insulin 2 h after feeding of a single dose of glucose (1 g/kg) may be a simple and practical way to aid identification of laminitis-prone ponies before the onset of clinical disease, particularly when ponies are adapted to eating fall pasture.     

Fermentation of six different forages in the semi-continuous fermentation technique Caesitec

The aim of the present study was to compare carbohydrate degradation of forages which store carbohydrates either predominantly as fructan or starch, in horses’ hindgut. The effects of an abrupt change from hay‐based feeding to green fodder‐based feeding on the caecal flora were tested with the in vitro hindgut simulation technique ‘Caesitec’. Six trials with different forages (English ryegrass, tall fescue, grass mixture‐horses, grass mixture‐cows, lucerne, white clover) were conducted. During a 4‐day stabilisation period, samples were taken once a day before loading the fermenters with hay. After diet‐change to forage‐based feeding, samples were taken four times a day. Ammonia and pH‐value were measured before and 1, 2 and 6 h after loading the ‘Caesitec’. Gas formation was measured daily. Bacterial numbers, lactate and short chain fatty acids were detected at four time‐points of each trial. The grass mixtures contained the highest amounts of fructan. The pH‐values were in the physiological range from pH 6 up to 7 (6.58–6.83) by feeding all forages. Gas formation, anaerobic and aerobic bacterial numbers increased after diet change from hay to any forage. The maximum amount of fructan (3.75 g/kg) in swiss pasture did not cause a permanent pathological change in the hindgut‐flora.     

Effects of excess caloric fat feeding on the lipid metabolism in Shetland ponies

To investigate the influence of overweight and dietary fat supplementation on lipid and insulin glucose metabolism of Shetland ponies, eight Shetland pony geldings were fed a hypercaloric (30 MJ DE/150 kg bwt. and day) fat diet (10% fat as soybean oil) or a carbohydrate control diet for nine months until ponies gained an overweight of 15%. Afterwards oral glucose tolerance tests (oGTT; 5, 6 mmol/kg bwt.) were performed after a 12 hour fast and after a fast which led to an increase of plasma triglyceride concentrations to a threshold of 3 mmol/l (36-65 hrs.). Plasma concentrations of glucose, insulin, triglycerides and non esterified fatty acids (NEFA) were determined for 480 minutes after the glucose load. Ponys having had received the control diet tended to a higher insulin secretion in case of both oGTTs, whereas the glucose tolerance was similar in both groups but lower than in ponies of normal weight. During the oGTTs after fasting leading to the plasma triglyceride threshold, triglyceride concentrations decreased significantly (p < 0.05) faster and stronger in fat fed ponies. Additionally, fat fed pony showed significantly (p < 0.05) lower NEFA levels. The results of this study demonstrate a positive effect of fat feeding on the triglyceride clearance of overweight Shetland ponies.     

Hypertension and insulin resistance in a mixed-breed population of ponies predisposed to laminitis

OBJECTIVE: To determine the metabolic phenotype of a group of laminitis-prone ponies when at pasture in summer, compared with when at pasture in winter. ANIMALS: 40 ponies of various breeds predisposed to recurrent pasture-associated laminitis and 40 unaffected control ponies. PROCEDURES: Body condition score and size of the crest of the neck were assessed, blood samples obtained, and blood pressure measured by use of an indirect oscillometric technique, while ponies were kept on winter pasture (last week of November or beginning of December) and again on summer pasture (June). Serum insulin concentration and plasma glucose, triglyceride, uric acid, and ACTH concentrations were measured. Insulin sensitivity was calculated with proxies derived from basal serum insulin and plasma glucose concentrations. RESULTS: No significant differences were apparent between ponies predisposed to laminitis and control ponies during winter. However, in June, laminitis-prone ponies had increased serum insulin concentration and plasma triglyceride and uric acid concentrations, compared with control ponies. Also, laminitis-prone ponies were relatively insulin resistant, compared with control ponies. Mean blood pressure was significantly higher during summer in laminitis-prone ponies (median [interquartile range], 89.6 mm Hg [78.3 to 96.9 mm Hg]), compared with control ponies (76.8 mm Hg [69.4 to 85.2 mm Hg]). CONCLUSIONS AND CLINICAL RELEVANCE: Summer pastures appear to induce metabolic responses in some ponies, leading to expression of the prelaminitic phenotype, which includes hypertension as well as insulin resistance. Signs of this metabolic syndrome may not be apparent in affected ponies during periods of grazing winter pasture. Understanding this syndrome may enable improved countermeasures to be devised to prevent laminitis.     

Distribution of insulin receptor and insulin‐like growth factor‐1 receptor in the digital laminae of mixed‐breed ponies: An immunohistochemical study

Reasons for performing study: Hyperinsulinaemia has been implicated in the pathogenesis of laminitis; however, laminar cell types responding to insulin remain poorly characterised. Objectives: To identify laminar cell types expressing insulin receptor (IRc) and/or insulin‐like growth factor‐1 receptor (IGF‐1R); and to evaluate the effect of dietary nonstructural carbohydrate (NSC) on their expression. Methods: Mixed‐breed ponies (n = 22) received a conditioning hay chop diet (NSC ～6%); following acclimation, ponies were stratified into lean (n = 11, body condition score [BCS]≤4) or obese (n = 11, BCS ≥7) groups and each group further stratified to remain on the low NSC diet (n = 5 each for obese and lean) or receive a high NSC diet (total diet ～42% NSC; n = 6 each for obese and lean) for 7 days. Laminar samples were collected at the end of the feeding protocol and stained immunohistochemically for IRc and IGF‐1R. The number of IRc(+) cells was quantified; distribution of IGF‐1R was qualitatively described. Laminar IRc content was assessed via immunoblotting. Results: The number of IRc(+) cells was greater in the laminae of high NSC ponies than low NSC ponies (P = 0.001); there was a positive correlation between the change in serum insulin concentration and number of IRc(+) cells (r²= 0.74; P<0.0001). No epithelial IRc(+) cells were observed; IRc(+) cells were absent from the deep dermis. Analysis of serial sections identified IRc(+) cells as endothelial cells. The distribution of IGF‐1R was more extensive than that of IRc, with signal in vascular elements, epithelial cells and fibroblasts. Conclusions: Increased dietary NSC results in increased laminar endothelial IRc expression. Laminar keratinocytes do not express IRc, suggesting that insulin signalling in laminar epithelial cells must be mediated through other receptors (such as IGF‐1R). Potential relevance: Manipulation of signalling downstream of IRc and IGF‐1R may aid in treatment and prevention of laminitis associated with hyperinsulinaemia.     

The occurrence of feed-induced stomach ulcers in horses

Ponies were fed hay (n = 21) or mixed feed (n = 27; 128 g crude protein, 175 g crude fibre/kg dry matter) for at least 14 days (treatment against parasites with ivermectin before the experimental period). The feeding level was 18 g dry mater/kg body weight/day. After the feeding period (the experiments were conducted to investigate the water and electrolyte content of the alimentary tract in dependence on feed, feeding time and physical exercise) the ponies were slaughtered and the stomachs were visually controlled for the occurrence of ulcers. In the hay group no gastric ulcers were observed, but from the 27 ponies given mixed feed 10 (chi 2 = 7.71) animals showed ulcers on the cutaneous mucosa of the pars oesophagea localized along the margo plicatus. At 3.5 hours postprandial the percentage of dry matter in gastric contents of concentrate fed ponies was higher than in those which consumed hay; 12 hours after feeding the opposite took place. The gastric fill was rather similar, while the amount of chlorine and the pH-value respectively were 37 (+/- 16) mg/kg BW and 3.83 (+/- 0.83; n = 8) respectively in hay fed animals compared to 45 (+/- 17) mg/kg BW and 4.84 (+/- 0.55; n = 11) respectively in ponies, which consumed the mixed feed.     

Glycaemic and insulinaemic responses to feeding hay with different non-structural carbohydrate content in control and polysaccharide storage myopathy-affected horses

The aim of this study was to determine whether the glycaemic/insulinaemic responses to hay with non-structural carbohydrate (NSC, soluble carbohydrate) of 17% (HC), 10% (MC) or 4% (LC) differs in control horses and whether these responses differ between control and horses with polysaccharide storage myopathy (PSSM). Five clinically normal control horses and seven PSSM horses, all unfit and of Quarter Horse breeding (age 9.4 ± 3.4 years, body condition score range: 4.5-6). A crossover design compared the HC and LC hay, with horses randomly assigned to hay type for 5 days, and all horses fed the MC hay during washout, after which the diets were switched. Horses were fed 1.5% BW (as fed) divided into 2 feeding per day, no grain. On morning of the fifth day of each block (seventh day for washout), horses were given 0.5% BW in hay, blood was drawn before and every 30 min for 5 h after feeding, and the rate of intake was measured. Whole blood glucose and plasma insulin were measured. The intake rate was significantly higher for HC. In control horses, the insulin area under the curve (6891.7 ± 3524.2 HC vs. 1185.4 ± 530.2 LC) was significantly higher than LC. Polysaccharide storage myopathy horses had significantly higher glycaemic and insulinaemic responses to HC vs. LC, however; the magnitude of insulin response was lower and glucose response higher in PSSM vs. control horses. Results suggest that insulin responses can differ significantly with the NSC content of hay. Feeding hay with 17% NSC produces elevations in insulin that could be detrimental for PSSM horses.     

Influence of high phosphorus intake on salivary and plasma concentrations,and urinary phosphorus excretion in mature ponies

This study addressed the question whether the concentration of phosphorus (P) in saliva of ponies is influenced by P intake. Six ponies were fed a diet high in P (HP treatment), providing 21 g P/day, and a diet low in P (LP treatment), supplying 7 g P/day. The two diets provided approximately 21 g calcium (Ca) and 6 g magnesium (Mg)/day. The experiment had an A‐B‐A design with treatment periods of 30 days. The ponies first received the HP diet (HP1), followed by the LP treatment and were then fed again the HP diet (HP2). Urinary P excretion was increased in both HP feeding periods and equalled approximately 7% of P intake vs. 0.5% on the LP diet. Plasma P concentration was higher for the HP treatment. The salivary P concentration ranged from 0 to 1.01 mmol P/l between ponies and there was no effect of P intake. It is suggested that saliva is not an important excretion route of P. The percentage of Ca and Mg in urine (% of intake) was higher for the LP treatment than for the HP treatments. The results of this study suggest that salivary Mg may contribute to Mg homeostasis.     

Gluconeogenesis from propionate produced in the colon of the horse

The production of propionate has been measured in the large colon of two ponies fitted with cannulas and fed on a standard diet of hay or hay and wheat bran. A continuous infusion of 14C-labelled sodium propionate was made into a cannula in the right ventral colon and samples of ingesta were obtained from another cannula near the end of the right dorsal colon. A simultaneous intravenous infusion of [2-3H]-labelled glucose was made to measure total glucose entry. Colonic propionate production on the hay diet was 146 (range 110-176) mg/h per kg body weight and, on the hay and bran, 195 (range 130-273) mg/h per kg body weight. Mean total glucose production for the two diets was 120 (range 98-143) and 92 (range 79-116) mg/h per kg body weight, respectively. The results obtained indicate that 50% of the glucose was synthesized from propionate produced in the colon in ponies on the hay diet and 61% in ponies on the hay and bran diet.     

Feeding practices associated with colic in horses.

OBJECTIVE: To determine whether specific feeding practices were associated with development of colic in horses. DESIGN: Prospective matched case-control study. ANIMALS: 364 horses examined by veterinarians in private practice in Texas because of colic (cases; n = 182) or any other reason (controls; 182). PROCEDURE: Participating veterinarians were sent forms at the beginning of the study to collect information on signalment, feeding management practices, farm management practices, and preventive medical treatments. Case and control horses were compared by use of conditional logistic regression to identify factors associated with colic. RESULTS: Risk factors for colic were a recent change in batch of hay, decreased exposure to pasture, a recent change in type of grain or concentrate fed, feeding > 2.7 kg (6 lb) of oats/d, feeding hay from round bales, and Thoroughbred breed. Recent anthelmintic administration decreased the risk of colic. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that certain changes in diet (eg, change in batch of hay, change in type of grain or concentrate, feeding hay from round bales) and management (eg, decreased availability of pasture) increase the risk of colic in horses.     

Serum protein changes in ponies on different parasite control programmes

Serum protein responses were examined in 52 ponies divided into five groups and subjected to various control strategies that resulted in pasture infectivity ranging from 706 to 18,486 infective third stage, cyathostome and Trichostrongylus axei larvae per kilogram of herbage (L3/kg) by 17 September 1984. Major protein changes occurred only in young ponies (Groups 4 and 5) and were observed before exposure to maximum numbers of pasture larvae (Group 4; 10,210 L3/kg, Group 5: 10,042 L3/kg) on 17 September. It appeared that a primary infection of T axei was a greater stimulus to serum beta-globulin and immunoglobulin (Ig)G(T) responses that provided by continued infection with cyathostome (small strongyle) worms. The large strongyles (Strongylus vulgaris, S edentatus and S equinus) were not detected in any larval cultures or on pastures grazed by the young ponies. A fall in beta-globulin and IgG(T) concentrations of Group 5 ponies one month after treatment with ivermectin indicated a larvicidal action against T axei and/or the cyathostomes. A subsequent rise in serum albumin concentrations of Group 5 ponies suggested that a protein-losing gastroenteropathy had been alleviated by the larvicidal action of ivermectin. Mature control ponies (Group 1) showed little beta-globulin response and only a modest IgG(T) response in six of the 10 ponies after exposure to heavily infected lawns (18,486 L3/kg) in September 1984. It was concluded that serum protein and IgG(T) responses were of limited value as an aid to diagnosis of parasitism because of numerous difficulties of interpretation.     

燕麦干草、青贮燕麦与天然牧草饲喂牦牛的营养价值比较研究

为比较四川牦牛常规粗饲料的营养价值,试验选用燕麦干草、青贮燕麦、天然牧草,对其进行营养成分和体外产气分析。体内消化试验于四川省红原县进行,选取9头初始体重为（150±10） kg的麦洼公牦牛,随机分为3组,每组3头牛。试验结果如下：1）天然牧草粗蛋白（crude protein, CP）含量显著高于青贮燕麦和燕麦干草（P<0.05）;天然牧草的中性洗涤纤维（neutral detergent fiber, NDF）和酸性洗涤纤维（acid detergent fiber, ADF）含量显著低于青贮燕麦（P<0.05）;2）康奈尔净碳水化合物蛋白质动态评价体系（cornell net carbohydrate and protein system, CNCPS）中,青贮燕麦的非蛋白氮（non-protein nitrogen, PA）部分显著高于燕麦干草和天然牧草（P<0.05）;天然牧草的快速降解蛋白（rapidly degraded protein, PB1）、慢速降解蛋白（slowly degraded protein, PB3）含量最高,且不可降解蛋白（undegraded protein, PC）含量最低;青贮燕麦的不可利用纤维（undegraded fiber, CC）含量显著高于天然牧草和燕麦干草（P<0.05）;可利用纤维（available fiber, CB2）、非结构性碳水化合物（non-structural carbohydrates, NSC）含量显著低于天然牧草和燕麦干草（P<0.05）;3）天然牧草的NDF、CP消化率均显著高于燕麦干草和青贮燕麦（P<0.05）;天然牧草ADF消化率显著高于青贮燕麦（P<0.05）,与燕麦干草差异不显著（P>0.05）;4）天然牧草牦牛瘤胃液的NH₃-N浓度显著高于青贮燕麦和燕麦干草组（P<0.05）,而其微生物蛋白质（microbial crude protein, MCP）产量显著高于燕麦干草组,与青贮燕麦组差异不显著;燕麦干草总挥发性脂肪酸（total volatile fatty acid, TVFA）产量最高;5）天然牧草组的产气量显著高于青贮燕麦组（P<0.05）,与燕麦干草组差异不显著（P>0.05）;体外降解方面,燕麦干草的体外干物质降解率（in vitro dry matter digestibility, IVDMD）、体外中性洗涤纤维降解率（in vitro neutral detergent fiber digestibility, IVNDFD）、体外酸性洗涤纤维降解率（in vitro acid detergent fiber digestibility, IVADFD）最高,其中,IVDMD显著高于青贮燕麦（P<0.05）,与天然牧草差异不显著（P>0.05）;IVNDFD、IVADFD与其他两种饲草相比差异均不显著（P>0.05）。综上,CNCPS分析的各组分含量所反映的瘤胃发酵状况、体外产气法所测定的结果与牦牛体内消化试验测定的各养分消化率和瘤胃代谢产物结果基本吻合, 3种饲草的营养价值由高到低依次为天然牧草>燕麦干草>青贮燕麦。     

Triglyceride, Insulin, and Cortisol Responses of Ponies to Fasting and Dexamethasone Administration

Ponies were evaluated for their response to feed withholding and exogenous administration of corticosteroids (dexamethasone 0.04 mg/kg intramuscular [IM]) in an attempt to reproduce the hyperlipemia syndrome. Because insulin resistance has been associated with hyperlipemia, all ponies were initially evaluated for insulin response to an oral glucose load and normal dexamethasone suppression of serum cortisol. Four ponies were identified as hyperinsulinemic reflecting insulin resistance. All ponies had suppressed cortisol concentrations following dexamethasone administration. Feed withdrawal resulted in hypertriglyceridemia by 48 hours in all ponies. Very low density lipoprotein-triglyceride (VLDL) fraction was primarily elevated. The administration of dexamethasone failed to increase the degree of triglyceridemia. Although insulin resistance has been proposed as the likely cause of the hypertriglyceridemia in ponies, in this study four of eight ponies were considered to have normal insulin responses and yet still developed hypertriglyceridemia.     

Energy and fibre intake in a group of captive giraffe (Giraffa camelopardalis) offered increasing amounts of browse

We investigated the effect of diet on intake of energy and fibre in a group of three captive adult giraffe by weighing offered diet items and leftovers for 7 days after an adaptation period of 7 days. Digestion coefficients were calculated using, as internal marker, the acid detergent lignin content of a faecal sample pooled from subsamples taken during the last 5 days of intake measurement. Two lucerne hay-only diets of differing quality (L1, L2) were fed, as well as the regular diet of lucerne hay and concentrates (L2C), and the regular diet supplemented with 3 or 6 kg of edible, fresh browse material (L2CB3, L2CB6). The proportion of roughage in the ingested diets L2CB6 (45 +/- 5% dry matter), L2CB3 (35 +/- 3%) and L2C (37 +/- 10%) did not differ significantly. Digestible energy intake was low on the hay-only diets [L1: 0.28 +/- 0.06 MJ/kg body weight (BW)0.75; L2: 0.33 +/- 0.10 MJ/kg BW0.75] and increased from L2C (0.60 +/- 0.13 MJ/kg BW0.75) to a significant increase with L2CB3 (0.72 +/- 0.17 MJ/kg BW0.75); no further increase was obvious for L2CB6 (0.63 +/- 0.15MJ/kg BW0.75). The results confirm that giraffes are unlikely to meet energy requirements on lucerne hay-only diets. In a feeding scenario where both lucerne hay and the concentrate component of the diet are fed ad libitum, the animals tended to exchange hay for browse when browse was added. Only the higher level of browse supplementation led to a potentially beneficial increase in fibre intake. Whether additional browse supplementation will lead to increased intakes in a feeding scenario with restricted concentrate provision can be suspected but remains to be demonstrated.     

Effects of fat feeding and energy level on plasma metabolites and hormones in Shetland ponies

The aim of this study was to investigate the influence of a fat-supplemented diet compared with a carbohydrate diet on the lipid metabolism and the enteroinsular axis of Shetland ponies. The 'crossover' experiment was divided into two parts: in the first 10 weeks the diets comprised the correct number of calories according to requirements and in the following 10 weeks they were hypercaloric, in order to check the effect of a different energy content of the diets. Feeding the fat-enriched diet, independently of its energy content, led to a significant decrease in plasma triglycerides, associated with a mean 50% increase of plasma lipoprotein lipase activity. After oral glucose load the ponies on fat-enriched diets showed higher plasma glucose concentrations. Oral glucose administration after feeding the hypercaloric fat-enriched diet led to a 25-fold increase of plasma insulin levels. Glucose-dependent insulinotropic polypeptide plasma levels were increased in the animals on the fat-enriched diets. The results of this study suggest that fat feeding improves triglyceride clearance. However, the fat supplementation of the diet also led to impaired glucose tolerance. These results are important for a better understanding of the function of the enteroinsular axis. To investigate the influences of fat on lipid metabolism in relation to the aetiopathogenesis of equine hyperlipaemia further studies involving diseased animals are needed.     

Hormonal patterns in normal and hyperleptinemic mares in response to three common feeding-housing regimens

We previously reported that a rise in plasma leptin concentrations followed the rise in insulin and glucose in meal-fed horses, whereas horses maintained on pasture had little fluctuations in hormonal patterns. We have also described a hyperleptinemic-hyperinsulinemic condition that occurs in about 30% of our light horse mares of high body condition maintained on pasture. The present experiment was designed to 1) study the effect of 3 common feeding-housing regimens on leptin and other metabolic hormones in mares and 2) determine whether the hyperleptinemic condition interacted with these regimens. Six light horse mares with high body condition (average score = 7) were assigned to 2 simultaneous 3 x 3 Latin squares, 1 with normal mares (leptin = 0.1 to 6 ng/mL) and 1 with mares displaying hyperleptinemia (>10 ng/mL). Three feeding-housing regimens were compared: ad libitum pasture, ad libitum native grass hay in an outdoor paddock, and single morning feedings of a pelleted concentrate and hay at 0700 in a barn. Five days of acclimation to the feeding regimens were followed by a 36-h period of hourly blood collection to characterize the hormonal characteristics. Leptin concentrations were elevated (P < 0.001) in mares predetermined to be hyperleptinemic compared with normal mares, regardless of the feeding regimen. Leptin was greatest (P < 0.01) in mares on pasture and least in mares fed hay. Variations over time (P < 0.01) were present for all hormones and metabolites studied. Glucose and insulin concentrations were greatest (P < 0.01) in mares on pasture, with meal-fed mares exhibiting an immediate rise in plasma concentrations of both after feeding. Mares on hay had low and constant concentrations of glucose, insulin, and leptin, with no apparent fluctuations. Cortisol, prolactin, and IGF-I did not differ with leptin status, whereas GH differed due to feeding-housing regimen (P < 0.02); there was also an interaction of leptin status and feeding-housing regimen for GH concentrations (P = 0.094). It was concluded that 1) estimates of hormonal secretion in horses based on frequent sampling, depending upon the hormone in question, can be profoundly affected by the feeding-housing regimens, and 2) the hyperleptinemic condition persists under differing conditions of feeding-housing.     

Effect of dexamethasone, feeding time, and insulin infusion on leptin concentrations in stallions

Three experiments tested the hypotheses that daily cortisol rhythm, feeding time, and/or insulin infusion affect(s) leptin secretion in stallions. Ten mature stallions received ad libitum hay and water and were fed a grain concentrate once daily at 0700. In Exp. 1, stallions received either a single injection of dexamethasone (125 microg/kg BW i.m.; n = 5) or vehicle (controls; n = 5) at 0700 on d -1. Starting 24 h later, blood samples were collected every 2 h for 36 h via jugular venipuncture. Cortisol in control stallions varied (P < 0.01) with time, with a morning peak and evening nadir; dexamethasone suppressed (P < 0.01) cortisol concentrations. Leptin and insulin were greater (P < 0.01) in the treated stallions, as was the insulin response to feeding (P < 0.01). Leptin in control stallions varied (P < 0.01) in a diurnal pattern, peaking approximately 10 h after onset of eating. This pattern of leptin secretion was similar, although of greater magnitude (P < 0.01), in treated stallions. In Exp. 2, five stallions were fed the concentrate portion of their diet daily at 0700 and five were switched to feeding at 1900. After 14 d on these regimens, blood samples were collected every 4 h for 48 h and then twice daily for 5 d. Cortisol varied diurnally (P = 0.02) and was not altered (P = 0.21) by feeding time. Insulin and leptin increased (P < 0.01) after feeding, and the peaks in insulin and leptin were shifted 12 h by feeding at 1900. In Exp. 3, six stallions were used in two 3 x 3 Latin square experiments. Treatments were 1) normal daily meal at 0700; 2) no feed for 24 h; and 3) no feed and a bolus injection of insulin (0.4 mIU/kg BW i.v.) followed by infusion of insulin (1.2 mIU.kg BW(-1).min(-1)) for 180 min, which was gradually decreased to 0 by 240 min; sufficient glucose was infused to maintain euglycemia. Plasma insulin increased (P < 0.01) in stallions when they were meal-fed (to approximately 150 microIU/mL) or infused with insulin and glucose (to approximately 75 microIU/mL), but insulin remained low (10 microIU/mL or less) when they were not fed. The increases in insulin were paralleled by gradual increases (P < 0.01) in leptin concentrations 3 to 4 h later in stallions fed or infused with insulin and glucose. When stallions were not fed, leptin concentrations remained low. These results demonstrate that feeding time, and more specifically the insulin increase associated with a meal, not cortisol rhythm, drives the postprandial increase in plasma leptin concentrations in horses.     

Effects of an intravenous endotoxin challenge on glucose and insulin dynamics in horses

OBJECTIVE: To evaluate the effects of endotoxin administered IV on glucose and insulin dynamics in horses. ANIMALS: 16 healthy adult mares. PROCEDURES: Each week of a 2-week randomized crossover study, each horse received an IV injection (duration, 30 minutes) of Escherichia coli O55:B5 lipopolysaccharide (LPS) in 60 mL of sterile saline (0.9% NaCl) solution (20 ng/kg) or sterile saline solution alone (control treatment). Frequently sampled IV glucose tolerance test procedures were performed at 24 hours before (baseline) and 24 and 48 hours after injection; glucose and insulin dynamics were assessed via minimal model analysis. RESULTS: 13 of 16 horses had a clinical response to LPS, which was characterized by mild colic and leukopenia. Before treatment, mean +/- SD insulin sensitivity was 2.9 +/- 1.9 x 10(4) L x min(1) x mU(1); this significantly decreased to 0.9 +/- 0.9 x 10(4) L x min(1) x mU(1) 24 hours after treatment (69% reduction) and was 1.5 +/- 0.9 x 10(4) L x min(1) x mU(1) 48 hours after treatment. At baseline, mean +/- SD acute insulin response to glucose was 520 +/- 196 mU x min x L(1); this significantly increased to 938 +/- 620 mU x min x L(1) (80% increase) and 755 +/- 400 mU x min x L(1) (45% increase) at 24 and 48 hours after LPS treatment, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Compared with baseline values, insulin sensitivity was decreased for 24 hours after IV injection of LPS, and affected horses had a compensatory pancreatic response. These disturbances in glucose and insulin dynamics may contribute to development of laminitis in horses.