The lipopolysaccharide model for the experimental induction of transient lameness and synovitis in Standardbred horses.

An established lipopolysaccharide (LPS) model previously described in Warmbloods, was inconsistent in Standardbred horses, where lameness was not detected despite the presence of synovitis. The present study aimed to determine the dose of LPS from E. coli O55:B5 required to induce mild to moderate lameness following middle carpal joint injection in Standardbred horses and to quantitate the induced lameness over time, with and without anti-inflammatory pre-treatment. In a baseline trial, eight healthy, clinically sound Standardbred horses were used in a rule-based dose-escalation design trial, starting at a dose of 10 endotoxin units (EU). Lameness at trot was evaluated visually and quantitatively (using an inertial-sensor system and pressure plate analysis). Synovial fluid aspirates were analysed for total nucleated cell counts, total protein and prostaglandin E2 (PGE2). Following 2 months wash-out, the effective LPS-dose determined in the baseline trial was used to evaluate the effect of anti-inflammatory treatment. A mixed model for repeated measures with horse as random effect was used for analysis. After injection of 10 EU LPS, the desired degree of lameness was observed in the baseline trial, with maximal lameness at post-injection hour (PIH) 4, followed by a rapid decline and return to baseline by PIH 48. No lameness was observed following pre-treatment with meloxicam. In synovial fluid, PGE2 was significantly higher at PIH 8 and PIH 24 in the baseline trial compared with following meloxicam pre-treatment. In conclusion, injection of the middle carpal joint with 10 EU LPS consistently induces a transient lameness and synovitis in Standardbred horses.

Dynamic evaluation of toe-heel and medio-lateral load distribution and hoof landing patterns in sound, unshod Standardbred horses with toed-in, toed-out and normal hoof conformation.

Dynamic load distribution and landing patterns play an important role in equine orthopaedics. The aim of this study was to analyse dynamic load distribution and hoof landing patterns of sound Standardbreds. Twenty-four sound, unshod Standardbreds were walked and trotted over a pressure plate to obtain the peak vertical force (PVF), vertical impulse (VI) and stance time (ST) of both forelimbs. Asymmetry indices between both forelimbs were calculated and the dynamic medio-lateral and toe-heel load distribution of the forelimbs were compared between normal, toed-in and toed-out horses. The hoof landing patterns were determined on the first loaded zone. Linear mixed models and Chi-square analysis were used for statistical analysis (α = 0.05). At walk and trot, there were no significant differences in asymmetry of PVF, VI and ST between normal (n = 9/24 horses), toed-in (n = 6/24 horses), and toed-out horses (n = 9/24 horses). In both forelimbs, a flat landing (39.6% and 70.8% at walk and trot, respectively) predominated over all hoof conformations. No significant association between hoof conformation and landing pattern was observed (P > 0.05). Toed-out horses had significantly higher loading of the medial zone at the end of the stance phase at walk than normal horses (P = 0.002). No other statistically significant differences in load distribution were observed (P > 0.05). In conclusion, flat landing was more common in Standardbred horses than previously described in Warmbloods, emphasising the need for breed-specific clinical reference data and biomechanical studies. Moreover, mild deviations in hoof conformation did not necessarily affect load distribution.

Dietary supplementation of micro-encapsulated sodium butyrate in healthy horses: effect on gut histology and immunohistochemistry parameters.

BACKGROUND: As colic and intestinal disorders are a major concern in horses, the aim of the present study was to investigate the effect of dietary supplementation of butyrate, known to have a diverse array of beneficial effects on intestinal health. The effect of micro-encapsulated sodium butyrate supplementation on gut histology and immunohistochemistry parameters was studied in 14 healthy warmblood horses destined for slaughter in two separate periods. Horses were fed a low fiber - high starch diet, designed to induce subsequent starch overflow in the large intestine, aiming to create a mild challenge for large intestinal health. Treatment included supplementation with either micro-encapsulated sodium butyrate (Excential Butycoat®, Orffa, Werkendam, the Netherlands) or placebo (containing only coating material). The horses were fed for 20 consecutive days at a dosage of 0.4 g/kg BW (body weight). At day 21, the horses were slaughtered and intestinal samples were collected for determination of gut pH, villus length, crypt depth and area % of CD3+ and CD20+ cells. RESULTS: Horses on the butyrate supplemented diet had significantly reduced crypt depths in the right dorsal colon compared to placebo-fed horses (P < 0.001). However, a treatment x period interaction (P = 0.002) was discovered regarding this parameter, which could not be explained by the authors. Further investigation into the number of KI67+ cells in the RDC crypts did not reveal any significant differences between treatments (P = 0.650), indicating that the reduction in crypt depth in butyrate-fed horses could not be explained by a significant difference in cellular proliferation. Intestinal pH, villus length and expression of intestinal CD3+ and CD20+ cells were not significantly affected by treatment at any intestinal level. CONCLUSIONS: Our data indicate that supplementation of micro-encapsulated sodium butyrate to the equine diet did not influence gut histology (with the exception of a decrease found in the crypts of the RDC) or immunohistochemistry parameters in healthy horses. Further research is warranted to investigate the impact of butyrate supplementation in horses with intestinal disease.

Domesticated equine species and their derived hybrids differ in their fecal microbiota.

BACKGROUND: Compared to horses and ponies, donkeys have increased degradation of dietary fiber. The longer total mean retention time of feed in the donkey gut has been proposed to be the basis of this, because of the increased time available for feed to be acted upon by enzymes and the gut microbiota. However, differences in terms of microbial concentrations and/or community composition in the hindgut may also underpin the increased degradation of fiber in donkeys. Therefore, a study was conducted to assess if differences existed between the fecal microbiota of pony, donkey and hybrids derived from them (i.e. pony × donkey) when fed the same forage diet. RESULTS: Fecal community composition of prokaryotes and anaerobic fungi significantly differed between equine types. The relative abundance of two bacterial genera was significantly higher in donkey compared to both pony and pony x donkey: Lachnoclostridium 10 and 'probable genus 10' from the Lachnospiraceae family. The relative abundance of Piromyces was significantly lower in donkey compared to pony × donkey, with pony not significantly differing from either of the other equine types. In contrast, the uncultivated genus SK3 was only found in donkey (4 of the 8 animals). The number of anaerobic fungal OTUs was also significantly higher in donkey than in the other two equine types, with no significant differences found between pony and pony × donkey. Equine types did not significantly differ with respect to prokaryotic alpha diversity, fecal dry matter content or fecal concentrations of bacteria, archaea and anaerobic fungi. CONCLUSIONS: Donkey fecal microbiota differed from that of both pony and pony × donkey. These differences related to a higher relative abundance and diversity of taxa with known, or speculated, roles in plant material degradation. These findings are consistent with the previously reported increased fiber degradation in donkeys compared to ponies, and suggest that the hindgut microbiota plays a role. This offers novel opportunities for pony and pony × donkey to extract more energy from dietary fiber via microbial mediated strategies. This could potentially decrease the need for energy dense feeds which are a risk factor for gut-mediated disease.

Multi-kingdom characterization of the core equine fecal microbiota based on multiple equine (sub)species.

BACKGROUND: Equine gut microbiology studies to date have primarily focused on horses and ponies, which represent only one of the eight extant equine species. This is despite asses and mules comprising almost half of the world's domesticated equines, and donkeys being superior to horses/ponies in their ability to degrade dietary fiber. Limited attention has also been given to commensal anaerobic fungi and archaea even though anaerobic fungi are potent fiber degrading organisms, the activity of which is enhanced by methanogenic archaea. Therefore, the objective of this study was to broaden the current knowledge of bacterial, anaerobic fungal and archaeal diversity of the equine fecal microbiota to multiple species of equines. Core taxa shared by all the equine fecal samples (n = 70) were determined and an overview given of the microbiota across different equine types (horse, donkey, horse × donkey and zebra). RESULTS: Equine type was associated with differences in both fecal microbial concentrations and community composition. Donkey was generally most distinct from the other equine types, with horse and zebra not differing. Despite this, a common bacterial core of eight OTUs (out of 2070) and 16 genus level groupings (out of 231) was found in all the fecal samples. This bacterial core represented a much larger proportion of the equine fecal microbiota than previously reported, primarily due to the detection of predominant core taxa belonging to the phyla Kiritimatiellaeota (formerly Verrucomicrobia subdivision 5) and Spirochaetes. The majority of the core bacterial taxa lack cultured representation. Archaea and anaerobic fungi were present in all animals, however, no core taxon was detected for either despite several taxa being prevalent and predominant. CONCLUSIONS: Whilst differences were observed between equine types, a core fecal microbiota existed across all the equines. This core was composed primarily of a few predominant bacterial taxa, the majority of which are novel and lack cultured representation. The lack of microbial cultures representing the predominant taxa needs to be addressed, as their availability is essential to gain fundamental knowledge of the microbial functions that underpin the equine hindgut ecosystem.

Evaluation of titanium dioxide and chromic oxide as digestibility markers in ponies fed alfalfa hay in relation to marker dosing frequency.

In equines, Cr2O3 is widely accepted as an indigestible marker, but there are health concerns regarding the carcinogenic properties of Cr2O3. Recently, TiO2 has been suggested to be an alternative digestibility marker in equines. However, a comparison between Cr2O3 and TiO2 has not been made in equines. Six Welsh pony geldings (initial BW: 254±3 kg; 7 years of age) fed chopped alfalfa hay were used to evaluate the use of TiO2 (Ti) and Cr2O3 (Cr) as markers for calculating apparent digestibility and to investigate the effect of frequency of marker administration on the measurement of digestibility values. Diets contained 4.65 kg dry matter (DM) chopped alfalfa hay supplemented with minerals, vitamins, TiO2 (3.3 g Ti/day) and Cr2O3 (3.2 g Cr/day). Ponies were dosed with either 3.3 g Ti and 3.2 g Cr once daily (DF1) or with 1.65 g Ti and 1.60 g Cr twice daily (DF2). After adaptation to the diets and procedures for 14 days, voluntary voided faeces were collected quantitatively over 7 days and analysed for moisture, ash, Ti and Cr. Apparent total tract DM digestibility (DMD) and organic matter digestibility (OMD) were calculated using the total faecal collection (TFC) and marker method (Ti and Cr). The overall mean cumulative faecal recovery of Cr and Ti (as % of intake) were 102.0% and 96.6%, respectively. Mean daily faecal recoveries of Cr as well as of Ti were not different (P=0.323; P=0.808, respectively) between treatments. Overall daily faecal recovery of Cr differed (P=0.019) from 100% when the marker was dosed once daily, whereas overall daily faecal recovery was similar to 100% for both administration frequencies when Ti was used as a marker. For both markers, the coefficient of variation of the mean faecal marker recovery between horses was lower when the markers were administrated twice per day. Across treatments, cumulative DMD and OMD estimated with Ti were similar (P=0.345; P=0.418, respectively) compared with those values determined by TFC method. When Cr was used, the calculated cumulative DMD tended (P=0.097) to be greater compared with those estimated with TFC, and cumulative OMD values were overestimated (P=0.013). Orally supplemented Ti recovery in the faeces of ponies fed chopped alfalfa hay with Ti administered once or twice daily was close to 100%, making it the preferred marker for digestibility trials in equines.

Effect of exercise on apparent total tract digestibility of nutrients and faecal recovery of ADL and TiO2 in ponies.

Exercise and physical training are known to affect gastrointestinal function and digestibility in horses and can lead to inaccurate estimates of nutrient and energy digestibility when markers are used. The effect of exercise on apparent nutrient digestibility and faecal recoveries of ADL and TiO2 was studied in six Welsh pony geldings subjected to either a low- (LI) or high-intensity (HI) exercise regime according to a cross-over design. Ponies performing LI exercise were walked once per day for 45 min in a horse walker (5 km/h) for 47 consecutive days. Ponies submitted to HI exercise were gradually trained for the same 47 days according a standardized protocol. Throughout the experiment, the ponies received a fixed level of feed and the daily rations consisted of 4.7 kg DM of grass hay and 0.95 kg DM of concentrate. The diet was supplemented with minerals, vitamins and TiO2 (3.0 g Ti/day). Total tract digestibility of DM, organic matter (OM), CP, crude fat, NDF, ADF, starch, sugar and energy was determined with the total faeces collection (TFC) method. In addition, DM and OM digestibility was estimated using internal ADL and the externally supplemented Ti as markers. Urine was collected on the final 2 days of each experimental period. Exercise did not affect apparent digestibility of CP, crude fat, starch and sugar. Digestibility of DM (DMD), OM (OMD), ADF and NDF tended to be lower and DE was decreased when ponies received the HI exercise regime. For all treatments combined, mean faecal recoveries of ADL and Ti were 87.8±1.7% and 99.3±1.7%, respectively. Ti was not detected in the urine, indicating that intestinal integrity was maintained with exercise. Dry matter digestibility estimated with the TFC, ADL and Ti for ponies subjected to LI exercise were 66.3%, 60.3% and 64.8%, respectively, while DMD for HI ponies were 64.2%, 60.3% and 65.2%, respectively. In conclusion, physical exercise has an influence on the GE digestibility of the feed in ponies provided with equivalent levels of feed intake. In addition, the two markers used for estimating apparent DMD and OMD indicate that externally supplemented Ti is a suitable marker to determine digestibility of nutrients in horses performing exercise unlike dietary ADL.

Ultrasonographic measurements of localized fat accumulation in Shetland pony mares fed a normal v. a high energy diet for 2 years.

Health risks associated with obesity are more likely a factor of the localization of fat excess, rather than of elevated BW per se. The aim of this randomized controlled clinical trial was to determine the effect of a long-term high energy diet on BW, fat accumulation and localization. Eight Shetland pony mares, 3 to 7 years old, were randomly divided into a control and a high energy (HE) diet group fed either maintenance or double maintenance energy requirements (200% net energy (NE)) for two consecutive summers, with a low energy diet in the winter in between. Body condition score (BCS) did not differ between the groups at the onset of the study (control 5.6±0.75 v. HE 6.3±0.5). From 12 weeks after starting the diet, ultrasonography of five different locations (retroperitoneal, axillary, withers, intercostal and rump) for adipose deposition, BCS and BW were measured monthly during the period that ponies received different diets. Statistical analysis was performed using a linear mixed-effects model with post hoc Bonferroni testing. P values <0.05 were considered significant. At week 12 after the onset of the diet, fat thickness in the HE group was significantly greater than in the control group. During the monitoring period, the HE group showed a significant increase in mean (±SE) BW (+52%, 265±13.94kg) and BCS (+70%; to 9.0±0.4), while the control group was unchanged (BW 160±13.98 kg; BCS 3.8±0.4). At all locations, the fat depth in the HE group increased significantly, with the highest increase noted for retroperitoneal deposits. The conclusions were that a 200% NE diet induced subcutaneous and retroperitoneal fat accumulation, with the greatest increase in intra-abdominal deposits. The moderate increase of the subcutaneous fat depth followed by a plateau phase suggests the existence of a limit of adipose tissue expandability, as in man.

Evaluation of methodological aspects of digestibility measurements in ponies fed different haylage to concentrate ratios.

Methodological aspects of digestibility measurements were studied in four Welsh pony geldings consuming haylage-based diets with increasing proportions of a pelleted concentrate according to a 4×4 Latin square design experiment. Ponies were fed four experimental, iso-energetic (net energy (NE) basis) diets (i.e. 22 MJ NE/day) with increasing proportions of a pelleted concentrate (C) in relation to haylage (H). The absolute amounts of diet dry matter fed per day were 4.48 kg of H (100H), 3.36 and 0.73 kg of H and C (75H25C), 2.24 and 1.45 kg of H and C (50H50C) and 1.12 and 2.17 kg of H and C (25H75C). Diets were supplemented with minerals, vitamins and TiO2 (3.7 g Ti/day). Voluntary voided faeces were quantitatively collected daily during 10 consecutive days and analysed for moisture, ash, ADL, acid-insoluble ash (AIA) and Ti. A minimum faeces collection period of 6 consecutive days, along with a 14-day period to adapt the animals to the diets and become accustomed to the collection procedure, is recommended to obtain accurate estimations on dry matter digestibility and organic matter digestibility (OMD) in equids fed haylage-based diets supplemented with concentrate. In addition, the recovery of AIA, ADL and Ti was determined and evaluated. Mean faecal recovery over 10 consecutive days across diets for AIA, ADL and Ti was 124.9% (SEM 2.9), 108.7% (SEM 2.0) and 97.5% (SEM 0.9), respectively. Cumulative faecal recovery of AIA significantly differed between treatments, indicating that AIA is inadequate to estimate the OMD in equines. In addition, evaluation of the CV of mean cumulative faecal recoveries obtained by AIA, ADL and Ti showed greater variations in faecal excretion of AIA (9.1) and ADL (7.4) than Ti (3.7). The accuracy of prediction of OMD was higher with the use of Ti than ADL. The use of Ti is preferred as a marker in digestibility trials in equines fed haylage-based diets supplemented with increasing amounts of pelleted concentrate.

The preventive effects of two nutraceuticals on experimentally induced acute synovitis.

BACKGROUND: Nutraceuticals are often used in the management of equine osteoarthritis, but scientific evidence of their efficacy is lacking. OBJECTIVES: To study the preventive effects of two new nutraceuticals after the experimental induction of synovitis in comparison with positive and negative control treatments. STUDY DESIGN: Blinded, controlled, randomised experiment. METHODS: Twenty-four healthy Standardbred horses were randomly allocated to supplement AT (multi-ingredient, 28 days), supplement HP (collagen hydrolysate, 60 days), meloxicam (4 days) or placebo (60 days). Synovitis was induced in the right intercarpal joint by intra-articular injection of 0.5 ng lipopolysaccharide (LPS) of Escherichia coli while treatments were continued. Blood and synovial fluid were sampled before treatment, immediately prior to LPS injection, and at 8, 24 and 48 h post-injection. Synovial fluid samples were analysed for total nucleated cell count (TNCC), total protein (TP) and selected biomarkers (prostaglandin E2 [PGE2 ], interleukin-6 [IL-6], glycosaminoglycans [GAGs], type II collagen synthesis [CPII], matrix metalloproteinase [MMP]). Lameness was scored by visual examination and pressure plate analysis immediately prior to LPS injection, and at 8, 24 and 48 h post-injection. Clinical examinations were performed before treatment, immediately prior to LPS injection, at 2, 4 and 6 h post-injection, and then twice per day during the test period. RESULTS: Before treatment and intra-articular challenge, there were no statistically significant differences among the treatment groups for any of the parameters. After intra-articular challenge, the placebo group showed significantly higher synovial fluid TP, TNCC and PGE2 compared with the meloxicam group, although the model did not induce a relevant amount of lameness. Both nutraceuticals resulted in significantly lower synovial fluid TP, TNCC and PGE2 compared with placebo. No statistical differences in IL-6, GAGs, CPII or MMPs were observed among treatment groups. No adverse effects were observed. MAIN LIMITATIONS: Despite evidence of synovitis, lameness was too mild to detect. CONCLUSIONS: The preventive administration of these nutraceuticals showed anti-inflammatory effects in this validated synovitis model. Therefore, further studies of their clinical applicability are warranted.

A commercially available immunoglobulin E-based test for food allergy gives inconsistent results in healthy ponies.

REASONS FOR PERFORMING STUDY: Commercial immunoglobulin E (IgE)-based tests are available for diagnosis of food allergies and are commonly used in equine practice. However, these tests have been proven unreliable as a screening method in man and other species, but not critically evaluated in equids. Therefore, a commercially available IgE-based test for horses was evaluated. OBJECTIVES: To evaluate the consistency of the results obtained with a commercially available IgE-based test for food allergy diagnosis in ponies (Phase I) and to subject ponies to a provocation trial with the presumed allergens (Phase II). STUDY DESIGN: Allergen screening followed by experimental food provocation trials in healthy ponies. METHODS: Blood samples of 17 healthy Shetland ponies were taken at 2 different time points, sent blinded to a commercial laboratory for screening of common food allergens and the results were evaluated for consistency (Phase I). Ponies that were positive for food allergens were consecutively challenged orally with each allergen separately for 14 days (Phase II). A washout period of one week was applied in ponies with multiple positive results. Clinical parameters and serum amyloid A were monitored during the provocation trial. RESULTS: Only 7/17 ponies were negative on the IgE-based test at the 2 time points, 3 had positive results twice but only one tested positive twice for the same food allergen. No abnormalities were noted during the provocation trials. CONCLUSIONS: This study demonstrated that this IgE-based test is not a reliable screening tool for food allergy in healthy equids.

Evaluation of methodological aspects of digestibility measurements in ponies fed different grass hays.

Methodological aspects of digestibility measurements of feedstuffs for equines were studied in four Welsh pony geldings consuming four grass-hay diets in a 4 × 4 Latin square design. Diets contained either a low (L), medium (M), high (H), or very high (VH) ADF content (264, 314, 375, or 396 g·kg DM, respectively). Diets were supplemented with minerals, vitamins, and TiO (3.9 g Ti·d). Daily feces excreted were collected quantitatively over 10 consecutive days and analyzed for moisture, ash, ADL, AIA, and titanium (Ti). Minimum duration of total fecal collection (TFC) required for an accurate estimation of apparent organic matter digestibility (OMD) of grass hay was assessed. Based on literature and the calculated cumulative OMD assessed over 10 consecutive days of TFC, a minimum duration of at least 5 consecutive days of fecal collection is recommended for accurate estimation of dry matter digestibility (DMD) and OMD in ponies. The 5-d collection should be preceded by a 14-d adaptation period to allow the animals to adapt to the diets and become accustomed to the collection procedures. Mean fecal recovery over 10 d across diets for ADL, AIA, and Ti was 93.1% (SE 1.9), 98.9% (SE 5.5), and 97.1% (SE 1.8), respectively. Evaluation of CV of mean fecal recoveries obtained by ADL, AIA, and Ti showed that variation in fecal Ti (6.8) and ADL excretion (7.0) was relatively low compared to AIA (12.3). In conclusion, the use of internal ADL and externally supplemented Ti are preferred as markers to be used in digestibility trials in equine fed grass-hay diets.

Daily methane production pattern of Welsh ponies fed a roughage diet with or without a cereal mixture.

Methane production from Welsh ponies fed 2 isoenergetic diets (NE basis) at maintenance was studied in a crossover design with 4 mature geldings (230 ± 10.5 kg BW, mean ± SE). Treatments included a roughage-only (R) diet (5.1 kg DM/d) or a roughage plus cereal mix (RC) diet (2.5 kg DM hay/d plus 1.1 kg DM cereal mix/d). For both diets, the same grass hay was used (898 g DM/kg and 4.5 MJ NE/kg DM) and a commercial cereal mix was used in the RC diet (890 g DM/kg and 9.6 MJ NE/kg DM). Ponies were housed in pairs in climate-controlled respiration chambers. Carbon dioxide production (CO2), oxygen (O2) consumption, and CH4 production were measured over 3 consecutive days. Heat production (HP) rates were calculated from gaseous exchange. Feces were collected quantitatively to determine dietary nutrient digestibility. Dry matter intake differed between diets (P < 0.0001), but NE intake was equal for both diets (22.3 ± 0.07 MJ NEm/d). Organic matter digestibility was lower (P = 0.006) for the R diet (47.2%) than the RC diet (55.6%). Methane production was higher (P = 0.014) on the R diet (29.8 L · pony(-1) · d(-1)) compared to the RC diet (23.2 L · pony(-1) · d(-1)). Methane production expressed in liters/kilogram metabolic body weight (BW0.75) per day tended (P = 0.064) to decrease with 21% for the RC group compared with the R group. Heat production, O2 consumption, and CO2 production were not affected by diet. Diurnal patterns of CH4 production and HP were similar for both diets. Methane production increased slightly (P < 0.652) after feeding and was numerically lower for the RC diet for all time points throughout the day. For both diets, HP was higher after feeding than before feeding and decreased again within approximately 3 h after feeding. Isoenergetic replacement of roughage by a cereal mix reduces CH4 production in ponies. No clear diurnal pattern in CH4 emission can be discerned in ponies fed at maintenance.

Repeated measurements of P retention in ponies fed rations with various Ca:P ratios.

This study addresses the question of whether feeding rations rich in P for a period of up to 42 d induces a positive P balance in adult ponies. Biochemical bone markers and parathyroid hormone (PTH; intact as well as whole PTH) were measured to obtain clues as to the effect of P loading on bone metabolism. The experiment had a Latin square design. Each feeding period lasted 42 d, and there were 2 balance trials (ECP1 and ECP2) within each feeding period. Each balance trial lasted 10 d (ECP1: d 11 to 21; ECP2: d 33 to 42). Six ponies aged 2.5 to 7 yr were fed a control diet that provided P and Ca according to the requirement (Control diet: 54 mg Ca·kg BW(-1) · d(-1); 36 mg P · kg BW(-1) · d(-1)), a diet high in Ca and P (HCaHP diet: 146 mg Ca · kg BW(-1) · d(-1); 121 mg P · kg BW(-1) · d(-1)), and a diet with a high P level only and Ca fed to the requirement (HP diet: 54 mg Ca · kg BW(-1) · d(-1); 122 mg P · kg BW(-1) · d(-1)). When fed the Control diet, the ponies showed a zero P and Ca balance over the 42-d period. The HCaHP diet resulted in both P and Ca retention (about 2 g Ca and P/d; P < 0.05). Phosphorus retention (about 2 g P/d) alone was observed when ponies were fed the HP diet, but P retention was only different (P < 0.05) from the Control diet in ECP1. The excretion of P in urine was reduced by greater Ca intake (P < 0.05), and Mg absorption was reduced by high P intake (P < 0.05). Plasma P concentration was raised by high P intake. Plasma Ca levels were not affected by dietary treatment. The greater (P < 0.05) P retentions observed for the HCaHP diet during ECP1 and ECP2 and HP diet during ECP1 could not be explained by processes that could have been indicated by the bone markers or PTH values. It was concluded that dietary-P-induced retention of P in ponies does not seem to be associated with altered bone metabolism in this study.

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.

The influence of calcium intake on phosphorus digestibility in mature ponies.

Literature data show conflicting results on the effect of feeding high calcium (Ca) levels on phosphorus (P) digestibility in ponies. None of the studies reported involved a dose response as to the effect of Ca intake on apparent P digestibility while keeping P intake constant. Thus, the influence of high dietary Ca level on apparent P digestibility was studied in balance trials by feeding six mature ponies three different Ca levels against a constant P intake. The ponies received 148, 316 and 535 mg Ca/kg BW/day with the low, intermediate and high Ca diet. Phosphorus intake was approximately 125 mg P/kg BW and magnesium (Mg) intake was approximately 35 mg Mg/kg BW/day. The intermediate and high Ca level decreased apparent digestibility, renal excretion and retention of P when compared with the low Ca level. However, apparent digestibility and renal excretion of P were not different between the intermediate and high Ca level. The high Ca level resulted in a retention of 66 mg Ca/kg BW/day. The percentage apparent digestibility of Ca was lower for the intermediate and high Ca level compared with the low Ca level. Magnesium digestibility was not affected by high dietary Ca level. In practice, high absolute amounts of Ca should be prevented especially when low amounts of P are fed; P digestibility will be depressed and a negative P balance may occur.

The apparent digestibility of phytate phosphorus and the influence of supplemental phytase in horses.

Availability of phytate-bound P as influenced by supplemental phytase was studied in eight horses consuming four diets in a 4 x 4 Latin square design experiment. The treatments were a control (containing a low P level, 18.4 g/d) and three high-P diets. These diets contained P as monocalcium phosphate (MCP; 43.7 g/d), myoinositol hexakisphosphate in the form of wheat and rice bran (MIHP; 41.8 g/d), or MIHP with microbial phytase (MIHPP; 42.5 g/d). The proportions of phytate-bound P were 3, 1, 55, and 56% for the control, MCP, MIHP, and MIHPP, respectively. The MIHPP diet was supplemented with 300 phytase units (FTU)/kg (as-fed basis). Feces and urine were collected quantitatively and analyzed for P, Ca, and Mg. Urinary P excretion was lower (P < 0.05) with the control diet (0 g of P/d) than with the MCP diet (1.0 g of P/d). The low urinary P excretion (0.3 g of P/d) for the MIHP diet suggested low P availability compared with the MCP diet, but apparent digestibility of P expressed as a percentage of intake did not differ (P = 0.065) between these diets. Apparent Ca digestibility was lower (P < 0.05) for the MIHP diet than for the MCP diet (26.4 vs. 42.4%). This difference may have been caused by the origin of the Ca in these diets. Phytase supplementation increased apparent Ca digestibility from 26.4 to 31.5% (P < 0.05). Magnesium was not influenced by the level of phytate in the diet. Our data indicate that phytase supplementation had more influence on Ca digestibility than on P digestibility and suggest that phytase supplementation may be beneficial for improving Ca digestibility for horses receiving a phytate-rich diet.

[Energy and protein supply for event horses during training: comparison between intake and requirements]. / De energie- en eiwitvoorziening van militarypaarden in training: vergelijking van opname en behoeftenormen.

In horses that exercise intensively (for example, event horses in training) the intake and energy requirements were compared on the basis of a diet record and estimates of energy required for exercise. Daily net energy intake over a 7 days period was on average 30% (n = 15) higher than the net energy requirement. Since the horses had a constant body weight, and thus were in energy balance, the energy intake was overestimated and/or the energy requirement was underestimated. The intake of digestible protein was 92% higher than the protein requirement. This study illustrates the problems concerning ration assessment and evaluation in practice.