Effects of β-Hydroxy-β-Methylbutyrate and γ-Oryzanol on Blood Biochemical Markers in Exercising Thoroughbred Race Horses

In both the horse and the man, nutritional ergogenic aids have been used to improve physical ability in conjunction with an appropriate training regimen. Although training increases physical condition, the ease of taking a nutritional additive to improve training results explains the demand for supplementation, which may increase mechanical energy of work, delay onset of fatigue, or improve neuromuscular coordination. The purpose of this study was to determine the effects of oral supplementation of β-hydroxy-β-methylbutyrate (HMB) and γ-oryzanol (GO) on indices of exercise-induced muscle damage in Thoroughbred race horses. In this 32-week study, the horses were assigned to either a placebo, GO (3.0 g/d), HMB (15 g/d), or GO and HMB treatment groups. The supplements were administered for the first 16 weeks of the study during the training period before the racing season began. Blood samples were taken at baseline, and then during training, before exercise, immediately after exercise, and 30 minutes after exercise. Heart rate and speed were monitored in each exercise session. Hematocrit, glucose, lactate (LA), creatine phosphokinase, and aspartate aminotransferase were measured before and after each exercise session. Analysis of variance showed a significantly greater increase in postexercise creatine kinase activity in placebo-supplemented group than in the other treatment groups, both in the training period and during the racing seasons (P < .05). Blood LA was higher immediately after exercise in the placebo group compared with the supplemented groups. In conclusion, supplementation with HMB and GO resulted in decreased creatine kinase and LA after exercise. These findings support the hypothesis that HMB and GO supplementation helps to prevent exercise-induced muscle damage.     

Atypical Radiologic Appearance of a Second Cervical Vertebral Fracture in a Horse

This report presents the case of a 21-year-old Warmblood mare referred to the clinic with neurological dysfunctions and considerable signs of discomfort. A fracture of the caudal vertebral body of C2 with a very unusual radiographic appearance was diagnosed, and the mare was euthanized due to deterioration in neurological status. Histology revealed no evidence of neoplasia or pathological fracture.     

Genetic relatedness and netB prevalence among environmental Clostridium perfringens strains associated with a broiler flock affected by mild necrotic enteritis

In a previous study we investigated pulsed-field gel electrophoresis (PFGE) genotype diversity and prevalence of the netB toxin gene in Clostridium perfringens (CP) isolates recovered from a broiler flock (flock 1) affected by necrotic enteritis (NE). In this follow-up work, we examined samples collected before placement of flock 1, to see if NE during rearing could be traced back to the cleaned and empty building or the day-old chicks. Litter from the next flock in the same building (flock 2) was also examined. We detected 25 different PFGE genotypes, five of which were found only in litter from flock 2. Six genotypes which had been found in flock 1 during rearing were detected in samples collected before placement. NetB positive isolates belonging to two of these genotypes had been recovered from NE lesions during rearing, suggesting that virulent strains were transmitted from the cleaned and disinfected broiler house. NetB frequency among isolates from the empty building was 45%, indicating that netB positive strains were prevalent in a building that previously had housed a healthy flock offered in-feed narasin (flock 0). NetB frequency among isolates from litter used by flock 2 was 22%, indicating that netB positive strains were present in the environment of a 14-days-old healthy flock offered in-feed narasin. Two prevalent genotypes were consistently either netB negative or netB positive. However, the presence of genotypes represented by both negative and positive isolates may suggest that the gene can spread horizontally among different CP strains.     

Iridoid, phenylethanoid and flavonoid glycosides from Sideritis trojana

From the MeOH extract of Sideritis trojana, a new iridoid glycoside, 10-O-(E)-feruloylmelittoside (1) was obtained in addition to four known iridoid glycosides [melittoside (2), 10-O-(E)-p-coumaroylmelittoside (3), stachysosides E (4) and G (5)]. Moreover, five phenylethanoid glycosides [verbascoside (6), isoacteoside (7), lamalboside (8), leonoside A (9), isolavandulifolioside (10), three flavone glycosides (isoscutellarein 7-O-[6'-O-acetyl-β-allopyranosyl-(1→2)]-β-glucopyranoside (11), 4'-O-methyisoscutellarein 7-O-[6'-O-acetyl-β-allopyranosyl-(1→2)]-β-glucopyranoside (12), 3'-hydroxy-4'-O-methyisoscutellarein 7-O-[6'-O-acetyl-β-allopyranosyl-(1→2)]-β-glucopyranoside (13) and a benzylalcohol derivative (di-O-methylcrenatin) were obtained and identified. The structures were elucidated on the basis of NMR and HRMS data. All compounds were tested for their antioxidant activity by in vitro TEAC assay and some of them exhibited moderate activity (0.97-1.44 mM) when compared with the reference compound (quercetin 1.86 mM). Glycosides 6-13, the most active compounds in the TEAC assay, were also tested by flow cytometry to evaluate their ability to affect the levels of reactive oxygen species (ROS) in human prostate cancer cells (PC3).     

Novel Prunus rootstock somaclonal variants with divergent ability to tolerate waterlogging

Plants require access to free water for nutrient uptake, but excess water surrounding the roots can be injurious or even lethal because it blocks the transfer of free oxygen between the soil and the atmosphere. Genetic improvement efforts in this study were focused on the increased tolerance in roots to waterlogging. Among a pool of clones generated in vitro from leaf explants of rootstock Mr.S.2/5 of Prunus cerasifera L., the S.4 clone was flood tolerant whereas the S.1 clone was sensitive. The S.4 clone formed adventitious roots on exposure to flooding. Moreover, the chlorophyll content and mitochondrial activity in the leaf and root, soluble sugar content, alcohol dehydrogenase activity and ethylene content were different between the clones. The sorbitol transporter gene (SOT1) was up-regulated during hypoxia, the alcohol dehydrogenase genes (ADH1 and ADH3) were up-regulated in the leaves and down-regulated in the roots of the S.4 clone during hypoxia, and the 1-aminocyclopropane-1-oxidase gene (ACO1) was up-regulated in the leaves and roots of the S.4 clone during hypoxia and down-regulated in the wild-type roots. In addition, in the S.4 root, hypoxia induced significant down-regulation of a glycosyltransferase-like gene (GTL), which has a yet-undefined role. Although the relevant variation in the S.4 genome has yet to be determined, genetic alteration clearly conferred a flooding-tolerant phenotype. The isolation of novel somaclonals with the same genomic background but with divergent tolerance to flooding may offer new insights in the elucidation of the genetic machinery of resistance to flooding and aid in the selection of new Prunus rootstocks to be used in various adverse environments.     

Antiproliferative steroidal glycosides from Digitalis ciliata

Two new compounds, a furostanol glycoside (1) and a pregnane glycoside (4), along with eight known compounds, belonging to the classes of spirostane (2,3), pregnane (5–7) and cardenolide (8–10) glycosides, were isolated from the seeds of Digitalis ciliata. Their structures were elucidated by 1D and 2D-NMR experiments as well as ESI-MS analysis. For the first time pregnane glycosides of the diginigenin series have been isolated from D. ciliata. The cytotoxic effects of compounds 1–10 on cell viability of several cancer cell lines, namely human breast cancer (MCF-7), human glioblastoma (T98G), human lung adenocarcinoma (A549), human colon carcinoma (HT-29), and human prostate cancer (PC-3) cell lines were evaluated. Compounds 1, 4, 7 and 8 showed antiproliferative effects against MCF-7, HT-29 and A549 cancer cells with IC₅₀ values ranging from 8.3 to 20 μM. The effects of compounds 1–10 on cell proliferation were evaluated on these three cancer cell lines by cell cycle analysis of DNA content using flow cytometry. Compounds 7, 8 and 10 induced significant changes in G₂/M cell cycle phase of all analyzed cells. The obtained results indicate that compounds 7, 8 and 10 are cytostatic compounds effective in reducing cell proliferation by inducing accumulation of the cells in the G₂/M phase of the cell cycle.     

The influence of temperature and age on the T1 relaxation time of the equine distal limb

The extent of fat suppression using short tau inversion recovery (STIR) imaging is variable between horses. Our aim was to determine if patient's age and/or hoof temperature have an influence on the T(1) relaxation time of bone marrow in the equine distal limb, thereby affecting the suppression of fat signal. Magnetic resonance imaging was conducted on standing horses and cadaver samples using a low-field magnet (0.27 T). The hoof temperature was measured at the lateral side of the coronary band. A modified inversion recovery fast spin-echo (IR-FSE) sequence was used to measure the signal intensity for a range of inversion times (TIs) at six different regions of interest (ROI): (1) distal aspect of the proximal phalanx, (2) proximal aspect of the middle phalanx, (3) distal aspect of the middle phalanx, (4) navicular bone, (5) proximal aspect of the distal phalanx, and (6) distal aspect of the distal phalanx. The T(1) of the bone marrow in the equine distal limb was calculated from the results and was found to increase by 3.13 ± 0.08 (SE) ms/°C. There was no significant effect of age (2-16 years) but the T(1) values measured from the limbs of young (< 1 year) animals were considerably longer (32.6 ± 1.7 (SE) ms). Similar effects of temperature and age were found for all measured ROIs but there were significant differences in the mean values of T(1) , ranging from +7.7 (distal aspect of the distal phalanx) to -13.2 ms (distal aspect of the proximal phalanx).     

Multicentric neurofibromatosis with rectal prolapse in a California sea lion (Zalophus californianus)

An approximately 31-yr-old California sea lion (Zalophus californianus) with a history of chronic visual impairment and corneal disease presented with slow onset, progressive neurologic deficits. Treatment for rear flipper paresis was not effective and the animal was euthanatized. Histopathologic findings included hepatocellular and biliary neoplasia, ocular amyloidosis, adrenal adenoma and pheochromocytoma, and spinal cord changes consistent with multicentric neurofibromatosis. This is the first documentation of these conditions in a California sea lion.     

Cardiac troponin I and the occurrence of cardiac arrhythmias in horses with experimentally induced endotoxaemia

Branding is the traditional and well-established method used to mark horses, but recently microchip transponders for implantation have become available. In this study, behaviour, physiological stress variables and skin temperature in foals were determined in response to hot-iron branding (n=7) and microchip implantation (n=7). Salivary cortisol concentrations increased in response to branding (1.8 ± 0.2 ng/mL) and microchip implantation (1.4 ± 0.1ng/mL), but cortisol release over time did not differ. In response to both manipulations there was a transient increase in heart rate (P<0.001) and heart rate variability (P<0.01). Branding and microchip implantation induced a comparable aversive behaviour (branding, score 3.86 ± 0.85; microchip, score 4.00 ± 0.82). Both techniques thus caused similar physiological and behavioural changes indicative of stress. Acutely, implantation of a microchip was as stressful as branding in foals. Branding caused a necrotising skin burn lasting at least 7 days. Moreover branding, but not microchip implantation (P<0.001), was accompanied by a generalized increase in skin temperature which was comparable to low degree post-burn hypermetabolism in humans.     

Effect of conventional and minimum tillage on physical and biochemical stabilization of soil organic matter

The objectives were to investigate (1) to which extent water-stable macro- and microaggregates sequester organic matter (OM) in a minimum tillage (MT) system compared to a conventional tillage (CT) system and (2) if the content of biochemically stabilized OM differs between both tillage systems, and (3) to study the temporal dynamics of the distribution of aggregate size classes and of storage of OM within aggregates in the field. Surface soils (0–5 cm) and subsoils (10–20 cm) were sampled after fallow (March 2007) and directly after tillage (November 2007) from a long-term experimental field near Göttingen, Germany. Macroaggregates (>0.25 mm) were in general less abundant after fallow than directly after tillage. In March, only 21% (CT) and 45% (MT) of C_org was stored within macroaggregates in the surface soil, whereas in November, the percentages increased to 58% and 73%, respectively. CT and MT soils of both depths were incubated as bulk soil (CT_bulk, MT_bulk) and with macroaggregates disrupted (<0.25 mm) (CT_md, MT_md) for 28 days at 22°C and water content of 50% of the maximum water holding capacity. For the MT_bulk and MT_md surface soils, C mineralization was significantly higher compared to the CT soils. Incubation of md soils did not generally result in a significantly higher C mineralization compared to the respective bulk soils, except for the MT_md subsoil. Acid hydrolysis showed that the proportion of biochemically stabilized, nonhydrolysable, C_org to total C_org was lower in the MT than in the CT soils. Overall, the data indicate that the effect of physical stabilization of OM stored in the macroaggregates may not be a mechanism protecting very labile C with a turnover time of weeks, but that longer preservation likely occurs after macroaggregate transformation into microaggregates, and the surplus of OM found in the surface soil of MT does not only depend on the biochemically stabilized OM. Finally, our data suggest that the temporal variability of distribution of aggregate size classes in the field is large, but spatial and operator variability also contributed to the observed differences.