Phosphorus excretion by mares post-lactation.

Across the equine literature, estimates of true P digestibility range from -23% to 79%. This large range cannot be explained by differences in P intake or phytate-P intake alone. However, differences in endogenous P secretion into the GI tract may explain the variation. In horses, excess absorbed P is not excreted in the urine but is re-secreted into the GI tract, increasing faecal P and leading to estimates of low P digestibility. Thus, accurate estimates of P digestibility can only be obtained if absorbed P is retained in the horse. The objective of this study was to examine P digestibility in post-lactational mares and control mares that were fed similar amounts of P. It was hypothesized that post-lactational mares would have greater P retention and higher apparent P digestibility than control mares. Prior to the study, four lactating and four non-lactating mares were fed a diet that provided 100% of the control mares' P requirement, but only 55% of the lactating mares' P requirement. During the study, both groups were fed P at the rate recommended for non-lactating mares. Post-lactational mares did not retain more P than control mares but tended to excrete more P than control mares (p = .082), presumably due to differences in endogenous P secretion into the GI tract. Metabolic changes occurring during mammary gland involution may have contributed to the increase in P excretion. However, faecal P excretion exceeded P intake in both groups (p = .08) and both groups lost weight during the study. Tissue mobilization during weight loss may have influenced P secretion into the GI tract.

Technical Note: A method for determination of titanium dioxide concentration in fecal samples.

Titanium dioxide has been used as a marker for determining diet digestibility indirectly, but some authors have expressed difficulty in measuring TiO2 concentrations in fecal material. We developed an accurate and precise method to determine TiO2 concentrations in equine feces. The method includes dry-ashing samples, digestion with (NH4)2SO4 in concentrated sulfuric acid, followed by the addition of H2O2 to produce a yellow to orange color that can be read spectrophotometrically. Accuracy was tested by spike recovery, and precision was tested by examining the coefficient of variation (CV) between duplicates of 449 individual samples. The method described here was compared with a previously published method by examining CV between duplicates of samples analyzed using both methods and comparing them using a paired t-test. Titanium dioxide spike recovery averaged 106%, and the CV between duplicates averaged 4.0%, with 79% of sample pairs having a CV of <5%. When compared with a previously published method, the method described here had a lower CV between duplicates (P < 0.0001). The method described here provides an accurate and precise quantitative analytical procedure for TiO2 in equine fecal samples.

Titanium dioxide is a marker fed to animals to help determine diet digestibility indirectly by measuring the concentration of TiO2 in fecal samples. This paper describes an accurate and precise method to analyze TiO2 in equine feces. The precision of this method is demonstrated by the low variation between sample duplicates. This method requires fewer sample replicates for analysis, leading to less labor, expense, and waste in the laboratory.

Interrelationships Between Age and Trace Element Concentration in Horse Mane Hair and Whole Blood.

The use of hair as a sample matrix to determine the mineral status of an animal has received a lot of interest. The objective of this study was to determine if the trace element content in horse mane hair changed with age when evaluated in a group of horses representing a large age range. As a second objective, whole blood trace element content was evaluated, and its relationship to mane hair trace element content, as well as age, were tested. Therefore, mane hair and whole blood samples were obtained from 59 horses, ranging from 2 months to 26 years in age, housed on the same farm. Mane hair samples were washed, and hair and blood digested and analyzed for 11 trace elements. Weak correlations (P < .05) between age and mane hair trace element content was detected for arsenic (r = 0.29), copper (r = -0.39) and selenium (r = -0.27). While the mane hair color did not affect trace element concentration, greater variability was detected in the black mane hair samples. This resulted in outliers that were removed from the final statistical analysis. However, data is presented to the reader both ways. Correlations were also detected (P < .05) between age and whole blood iron (r = 0.62), selenium (r = 0.76) and zinc (r = 0.47). This is similar to what has been reported in horses using serum or plasma. The trace element concentrations of mane hair and whole blood were not correlated (P > .05) in this study. However, the concentrations of trace elements in mane hair were higher than that of blood. Trace elements, specifically chromium and lead, that were below detection levels in the blood, were detectable in mane hair. This suggests that mane hair may be a potential means to investigate suspected exposure to excessive levels of trace minerals or heavy metals that are difficult to detect in blood.

Relationships Between Measurements of Body Fat in Thoroughbred Horses.

Equine obesity is increasing in prevalence, and weight loss diets are frequently recommended for these horses. However, there are also management situations in which horses are deemed to be too thin. To monitor the efficacy of weight change programs, estimates of body fat are often made. There are several systems available to estimate body fat, and there are benefits and challenges to using each method. The objective of this study was to compare four different methods of estimating body fat in Thoroughbred horses. In 14 mature Thoroughbred horses, relationships among body condition score (BCS), morphometric measurements, ultrasonic measures of subcutaneous fat depots, and estimation of total body fat (BFD) via measurement of total body water through deuterium oxide dilution were evaluated. Body condition scores ranged from 4.5 to 6.5 on a 9-point scale. Body condition score, heart girth-to-body weight ratio, and BFD were all positively correlated with each other (P < .05). Subcutaneous fat depth at the tailhead tended to be positively related to BFD when only horses with BCS ≥ 5 were included (P = .0680). These data suggest that BCS remains a simple means of monitoring adiposity in mature horses in moderate condition. Tailhead fat depots may become useful for monitoring changes in body fat in Thoroughbreds with a BCS above 5, although more work with animals of higher adiposity is required and at different times of year.

Effect of maternal diet on select fecal bacteria of foals.

Adult horses depend on the microbial community in the hindgut to digest fiber and produce short-chain fatty acids that are use for energy. Colonization of the foal gastrointestinal tract is essential to develop this symbiosis. However, factors affecting colonization are not well understood. The objectives of this study were to evaluate the age-related changes and effects of maternal diet on select fecal bacterial groups in foals from 1 to 28 d of age. Thoroughbred foals (n = 18) were from dams fed forage and one of two concentrates: an oat-based (OB) or corn and wheat middlings-based (CWB) pelleted concentrate. The mares had access to assigned concentrates, along with a mixed hay and cool-season grass pasture, 28 d before and 28 d after parturition. Fecal samples were collected from foals at 1 d (14 to 36 h), 4, 14, and 28 d after birth. Fecal samples were serially diluted with phosphate-buffered saline before inoculation of enriched, selective media to enumerate Lactobacillus spp., amylolytic bacteria, and cellulolytic bacteria. Enumeration data were log-transformed then analyzed with mixed model analysis of variance with repeated measures (SAS 9.3) to test the main effects of maternal diet (OB or CWB), time of sample, and interaction between maternal diet and time. Cellulolytic bacteria first appeared in foal feces between 4 and 14 d of age and increased with age (P < 0.05). Amylolytic bacteria and lactobacilli were abundant at 1 d and then increased with age (P < 0.05). There was an interaction between maternal diet and time for Lactobacillus spp. with OB foals having more lactobacilli than CWB foals at 1 and 4 d (P < 0.05); however, there were no differences observed at 14 d (P > 0.05). Maternal diet did not influence amylolytic or cellulolytic bacteria (P > 0.05). These results indicate that colonization of the hindgut is a sequential process beginning early in the foal's life and that maternal diet may influence some bacteria in the gastrointestinal tract of foals.

Effect of Starch Source in Pelleted Concentrates on Fecal Bacteria in Prepartum and Postpartum Mares.

Dietary starch source has been shown to affect fecal bacterial communities of horses fed minimally processed cereal grains. However, processing may increase foregut starch digestibility, reducing effects of starch source on fecal bacterial communities. This study aimed to determine the effect of starch source in pelleted concentrates on fecal Lactobacillus spp., amylolytic bacteria, and cellulolytic bacteria in broodmares mares, during the prepartum and postpartum period. Thoroughbred mares (n = 18) were paired by last breeding date then randomly assigned to either an oat-based or a corn and wheat middlings-based pelleted concentrate fed with forage. Mares were fed their assigned concentrates beginning on 310 days of gestation, and fecal samples were collected at 324 days of gestation, before parturition, 1 day, 14 days, and 28 days postpartum. Fecal samples were enumerated by serial dilution and inoculation into selective, enriched media for Lactobacillus spp., amylolytic bacteria, and cellulolytic bacteria. Data were log transformed then analyzed using a mixed model ANOVA with repeated measures (SAS 9.3) to test the main effects of treatment, time of sample, and treatment by time interaction. Starch source did not affect enumerated bacterial communities (P > .05); thus, pelleting concentrates may alter some of the effects of starch sources on the hindgut microbiota. Sample date did not affect amylolytic bacteria (P > .05); however, lactobacilli and cellulolytic bacteria decreased 1 day postpartum (P < .05). Although we did not observe an effect of starch source on fecal bacteria in mares, parturition did appear to alter the hindgut microbiota.

Effect of Dietary Starch Source and Concentration on Equine Fecal Microbiota.

Starch from corn is less susceptible to equine small intestinal digestion than starch from oats, and starch that reaches the hindgut can be utilized by the microbiota. The objective of the current study was to examine the effects of starch source on equine fecal microbiota. Thirty horses were assigned to treatments: control (hay only), HC (high corn), HO (high oats), LC (low corn), LO (low oats), and LW (low pelleted wheat middlings). Horses received an all-forage diet (2 wk; d -14 to d -1) before the treatment diets (2 wk; d 1 to 14). Starch was introduced gradually so that horses received 50% of the assigned starch amount (high = 2 g starch/kg BW; low = 1 g starch/kg BW) by d 4 and 100% by d 11. Fecal samples were obtained at the end of the forage-only period (S0; d -2), and on d 6 (S1) and d 13 (S2) of the treatment period. Cellulolytics, lactobacilli, Group D Gram-positive cocci (GPC), lactate-utilizers and amylolytics were enumerated. Enumeration data were log transformed and analyzed by repeated measures ANOVA. There were sample day × treatment interactions (P < 0.0001) for all bacteria enumerated. Enumerations from control horses did not change during the sampling period (P > 0.05). All treatments except LO resulted in increased amylolytics and decreased cellulolytics, but the changes were larger in horses fed corn and wheat middlings (P < 0.05). Feeding oats resulted in increased lactobacilli and decreased GPC (P < 0.05), while corn had the opposite effects. LW had increased lactobacilli and GPC (P < 0.05). The predominant amylolytic isolates from HC, LC and LW on S2 were identified by 16S RNA gene sequencing as Enterococcus faecalis, but other species were found in oat fed horses. These results demonstrate that starch source can have a differential effect on the equine fecal microbiota.