Mean retention time of digesta in the different segments of the equine hindgut

Six Thoroughbred horses were used to determine the mean retention time (MRT) of digesta in the different segments of the hindgut. The horses were fed timothy hay or silage in equal amounts (1.6% bodyweight [BW]/day) every 3 h/day. Hay or silage labeled with the rare earth elements cerium, dysprosium, neodymium, ytterbium, lanthanum, samarium and praseodymium, were fed to the horses 36, 30, 24, 18, 12, 6 and 3 h before slaughter, respectively. Just after slaughter, digesta samples in the different segments were collected and determined for the contents of the rare earth elements. Using these data, a method to calculate the MRT in the different segments was proposed. There were no significant differences between diets in the MRT of each segment. The averaged MRT of hay and silage in the cecum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon and small colon was 2.9, 3.1, 5.9, 1.0, 4.0, and 4.0 h, respectively. The dry matter weight of the digesta was related to the MRT in the right ventral colon (r = 0.94, P = 0.005), left ventral colon (r = 0.87, P = 0.03), left dorsal colon (r = 0.98, P < 0.001) and right dorsal colon (r = 0.95, P = 0.004), but it was not related to the MRT in the cecum and small colon.     

Phylogenetic analysis of hindgut microbiota in Hokkaido native horses compared to light horses

The analysis of 16S rDNA sequence of bacteria in feces of Hokkaido native horses and light horses were performed to compare the hindgut microbiota between the two breeds. One hundred and four bacterial 16S rDNA clones (57 clones from four native horses and 47 clones from two light horses) were obtained. Only four sequences (3.8% of total sequences) showed 97% or more similarity to known species. The sequences were mainly affiliated with Cytophaga–Flavobacter–Bacteroides and low GC Gram‐positive bacteria (LGCGP). Proportion of LGCGP was higher in light horses. Other phyla including Verrucomicrobia, Spirochaetes and Archaea were detected only for native horses, suggesting high diversity of microbiota in native horses. In LGCGP, clusters related to known cellulolytic species were found only for native horses, while a cluster related to soluble sugar‐utilizing species was detected only for light horses. The library composition‐comparing software LIBSHUFF showed significant (P < 0.05) difference of fecal microbiota between the horse breeds. The number of Fibrobacter succinogenes‐related sequence and the frequency of detection of novel groups were found to be higher in native horses by selective amplification analysis. The results suggest that genetic diversity and population size of the F. succinogenes group are higher in the hindgut of native horses.     

Maintenance of mouse trophoblast stem cells in KSR-based medium allows conventional 3D culture

Mouse trophoblast stem cells (TSCs) can differentiate into trophoblast cells, which constitute the placenta. Under conventional culture conditions, in a medium supplemented with 20% fetal bovine serum (FBS), fibroblast growth factor 4 (FGF4), and heparin and in the presence of mouse embryonic fibroblast cells (MEFs) as feeder cells, TSCs maintain their undifferentiated, proliferative status. MEFs can be replaced by a 70% MEF-conditioned medium (MEF-CM) or by TGF-ß/activin A. To find out if KnockOut^TM Serum Replacement (KSR) can replace FBS for TSC maintenance, we cultured mouse TSCs in KSR-based, FBS-free medium and investigated their proliferation capacity, stemness, and differentiation potential. The results indicated that fibronectin, vitronectin, or laminin coating was necessary for adhesion of TSCs under KSR-based conditions but not for their survival or proliferation. While the presence of FGF4, heparin, and activin A was not sufficient to support the proliferation of TSCs, the addition of a pan-retinoic acid receptor inverse agonist and a ROCK-inhibitor yielded a proliferation rate comparable to that obtained under the conventional FBS-based conditions. TSCs cultured under the KSR-based conditions had a gene expression and DNA methylation profile characteristic of TSCs and exhibited a differentiation potential. Moreover, under KSR-based conditions, we could obtain a suspension culture of TSCs using extracellular matrix (ECM) coating-free dishes. Thus, we have established here, KSR-based culture conditions for the maintenance of TSCs, which should be useful for future studies.     

Differences in grazing behavior of horses and cattle at the feeding station scale on woodland pasture

In this study, grazing behavior of mares and cows was compared on woodland pastures, in the context of foraging hierarchy. A horse‐group (12 native Hokkaido horses) and a cattle‐group (5 Hereford cows) were grazed on the woodland pastures of 13.3 ha and 5.8 ha, respectively. They grazed mainly on Sasa nipponica. Grazing behavior at a feeding station (FS) scale was recorded for three focal animals in each group. Mares took one bite per FS at a high percentage of FSs (24.3%) and cows took one, two and three bites per FS at high percentage (20.3%, 19.2% and 15.4%, respectively). Although feeding patch (FP) scale for both animals could be defined as a group of FSs, the length of FP and grazing behavior at FP scale were similar for both animals. The number of steps taken between two adjacent FSs or FPs for mares was larger than that for cows. The results suggested that both horses and cattle had similar foraging hierarchy from bite to FP scale but grazing behavior of horses at the FS scale was different from that of cattle, suggesting that grazing behavior of large herbivores has greater similarity in larger foraging hierarchies.     

Effects of grazing on deposition of chemical body components, energy retention and plasma hormones in steers

The objective of this study was to evaluate the effects of grazing on the accretion of energy and chemical components in steers, using the comparative slaughter technique and taking into consideration the growth rate. Twelve Holstein steers aged 6 months and weighing 160 kg were divided into three treatment groups – an initial, grazing and control group. The initial group was slaughtered at the beginning of the experiment. The grazing group was grazed on timothy pastures for 18 weeks. The control group was kept in a stable and offered concentrate and timothy hay in a proportion of 2:1 throughout the 18 weeks. The amount of ration offered to the control group was adjusted in order to obtain a growth rate similar to that of the grazing group. The grazing and control groups were slaughtered at the end of the experiment. The grazing and the control groups had similar growth rates of approximately 0.8 kg/day throughout the experimental period and similar final live weights of approximately 260 kg. The grazing group had lower fat and energy deposition rates than the control group (P < 0.05). Deposition rates of protein, fat, ash, moisture and energy were 130, 32, 30, 374 g/day and 4.4 MJ/day, respectively, in the grazing group and 117, 85, 32, 381 g/day and 6.2 MJ/day, respectively, in the control group. The net energy for growth was lower in the grazing group (7.7 MJ/kg) than in the control group (9.9 MJ/kg, P < 0.05). The grazing group had higher plasma concentrations in growth hormone and lower plasma concentrations in insulin and insulin‐like growth factor 1 than those in the control group (P < 0.05).     

Fiber digestion in various segments of the hindgut of horses fed grass hay or silage

We studied in vivo fiber digestibility and fermentation parameters such as volatile fatty acid concentration and in vitro degradation in each segment of hindgut of horses fed timothy hay or silage made from the same sward. Six Thoroughbred horses were fed timothy hay or silage in equal amounts (1.6% of bodyweight per day, bodyweight mean 572 kg) every 3 h per day, then slaughtered. There were no differences between hay and silage in the concentration of the total volatile fatty acids, the apparent digestibility of dry matter, organic matter, and fiber, and in vitro neutral detergent fiber (NDF) disappearance rate in each segment. The total volatile fatty acid concentration and the apparent digestibility of dry and organic matter and fiber differed ( P  < 0.01) depending on the digestive segments. Both for hay and silage, the concentration of total volatile fatty acids in the digesta liquid phase largely increased ( P  < 0.01) from the cecum to the right ventral colon, and kept a constant value from the right ventral colon to the right dorsal colon, and then decreased from the right dorsal colon to the small colon. For dry and organic matter and fiber components, the apparent digestibility changed in the same manner from former to hinder segments of the hindgut. Regardless of diet, dry and organic matter and fiber components showed lower values in the cecum and the right ventral colon and increased ( P  < 0.01) largely from the right ventral colon to the left dorsal colon, then kept constant values in segments to the rear of left dorsal colon. There were no differences in in vitro NDF disappearance among cecum, ventral colon and dorsal colon.