Specific polyphenol oxidase activity of red clover (Trifolium pratense) and its relation with forage quality in field experiments

Abstract

In feeding studies, red clover (RC) influenced positively the N utilization by ruminants. A relationship between polyphenol oxidase (PPO) activity and forage quality has not been established. Our objective was to investigate seasonal, site, genotype, and management effects on specific PPO activity in RC in three experiments under field conditions, and relate the activity to forage-quality parameters. In Experiment 1, six RC genotypes at two study sites were submitted to a 3-cut system. Specific PPO activity, forage quality, and vegetation stage were determined. PPO activity varied between harvest and study sites, with genotypes differing up to 3-fold in PPO activity within harvests. The specific PPO activity, forage quality, and vegetation stage in RC subjected to 5-cut system and grazing (Experiment 2) were determined. Additionally, in Experiment 3, cutting frequency in RC swards including mechanical stress (rolling) was investigated. The induction of PPO activity in RC by grazing or mechanical stress (Experiments 2 and 3) increased the activity up to 2.5-fold compared with RC at similar vegetation stage submitted to the 5-cut system. Mechanical stress induced by grazing or rolling, and seasonal differences, seem to have a larger influence on specific PPO activity than does the genotype effect observed in Experiment 1. For forage quality, an increased specific PPO activity explained 29–46% of the reduction in protein fraction ‘A’ content (non-protein N) in the cutting systems in Experiments 2 and 3. Other CP fractions achieved a lower relation. Furthermore, the precipitation-to-temperature ratio preceeding a harvest explained 63% of the variation in the specific PPO activity. In conclusion, the PPO activity in RC is induced by grazing and rolling. Whereas weather conditions preceeding a harvest showed a large influence, genotype influence had only minor relevance. These results may have implications for regional harvest management towards efficient N utilization by ruminants.     

Diagnosis of Taxus (yew) poisoning in a horse.

A 2-year-old bay Thoroughbred colt was found dead overnight in its stall without a known history of any illness, existing disease, or toxicant exposure. No information on the clinical signs before this animal's death was reported. A full necropsy was performed the next morning and revealed a mild to moderate degree of endocardial hemorrhages in both ventricles. Microscopic examination of the heart showed an acute mild multifocal necrosis of papillary muscles and ventricles. The stomach content contained approximately 2% Taxus alkaloids as determined by gas chromatography/mass spectrometry. In the past, diagnosis of Taxus poisoning has been mainly based on history of exposure and the presence of plant parts in the gastrointestinal tract. Pathological lesions associated with Taxus poisoning have not been published for horses. Therefore, this is the first report of cardiac lesions in a horse after lethal exposure to Taxus. On the basis of these findings, it is suggested that Taxus exposure needs to be considered in the differential diagnosis of horses that die suddenly or have cardiac lesions suggestive of Taxus exposure, even if intact plant parts are not identified in the stomach by the naked eye.     

Efficacy of milbemycin oxime in the treatment of canine generalized demodicosis: a retrospective study of 99 dogs (1995-2000)

Ninety-nine dogs diagnosed with generalized demodicosis were treated with milbemycin oxime. Cases diagnosed before two years of age were classified as juvenile-onset (53%) and those diagnosed after two years of age as adult-onset (47%). Dogs were considered cleared of mites when none could be demonstrated in scrapings and cured if no relapse was seen for 12 months. Eighty-five per cent (84/99) were cured with milbemycin oxime used for 1-6 months (mean 2.3 months) at a dosage of 0.5-1.6 mg kg-1 body weight (mean 0.75 mg kg-1). No significant difference in dosage or treatment time was seen between juvenile and adult cases. Chance of cure was significantly better in young animals (cured cases mean age 2.97 years) than in older animals (not cured cases mean age 8.02 years). Start of treatment early in the course of disease gave a significantly better chance of cure. Cases with severe pododemodicosis had less chance of a cure (9/11 not cured).     

Assessment of melamine and cyanuric acid toxicity in cats.

The major pet food recall associated with acute renal failure in dogs and cats focused initially on melamine as the suspect toxicant. In the course of the investigation, cyanuric acid was identified in addition to melamine in the offending food. The purpose of this study was to characterize the toxicity potential of melamine, cyanuric acid, and a combination of melamine and cyanuric acid in cats. In this pilot study, melamine was added to the diet of 2 cats at 0.5% and 1%, respectively. Cyanuric acid was added to the diet of 1 cat at increasing doses of 0.2%, 0.5%, and 1% over the course of 10 days. Melamine and cyanuric acid were administered together at 0%, 0.2%, 0.5%, and 1% to 1 cat per dose group. No effect on renal function was observed in cats fed with melamine or cyanuric acid alone. Cats dosed with a combination were euthanized at 48 hours after dosing because of acute renal failure. Urine and touch impressions of kidneys from all cats dosed with the combination revealed the presence of fan-shaped, birefringent crystals. Histopathologic findings were limited to the kidneys and included crystals primarily within tubules of the distal nephron, severe renal interstitial edema, and hemorrhage at the corticomedullary junction. The kidneys contained estimated melamine concentrations of 496 to 734 mg/kg wet weight and estimated cyanuric acid concentrations of 487 to 690 mg/kg wet weight. The results demonstrate that the combination of melamine and cyanuric acid is responsible for acute renal failure in cats.     

A high-throughput glasshouse bioassay to detect crown rot resistance in wheat germplasm

A high-throughput and reliable seedling bioassay to screen wheat germplasm for crown rot resistance was developed. Single wheat seedlings were grown in square seedling punnets in a glasshouse and inoculated with a monoconidial Fusarium pseudograminearum isolate 10 days after emergence. The punnets were laid horizontally on their side and a 10- µ L inoculum droplet placed on the stem base. Seedlings were incubated at near-saturated relative humidity, and crown rot severity was assessed 35 days after inoculation. Studies on the duration of incubation period, inoculum concentration and temperature were carried out to optimize these parameters. Seedling growth at 25/15(±5)°C in a glasshouse and 48-h incubation at near-saturated RH in darkness gave the best results. When crown rot resistance rankings of 16 Australian cultivars from the bioassay were compared with their field performance, Spearman's rank correlation coefficient was highly significant. This indicated that the seedling bioassay mimicked field resistance to crown rot in adult plants. A bootstrap resampling analysis showed little or no improvement in the coefficient of variation with an increasing number of replications, indicating a high level of precision and reproducibility. By detecting small but consistent differences in crown rot severity, the bioassay proved effective in large-scale screening for partial resistance: already over 1400 wheat genotypes have been screened. The high degree of precision makes this an invaluable tool in the understanding of pathogen aggressiveness, host specialization and parasitic fitness.     

Changes in compact bone microstructure of rats subchronically exposed to cadmium

Background

Chronic exposure to cadmium (Cd), even at low concentrations, has an adverse impact on the skeletal system. Histologically, primary and secondary osteons as basic structural elements of compact bone can also be affected by several toxicants leading to changes in bone vascularization and mechanical properties of the bone. The current study was designed to investigate the effect of subchronic peroral exposure to Cd on femoral bone structure including histomorphometry of the osteons in adult male rats.In our study, 20 one-month-old male Wistar rats were randomly divided into two experimental groups. In the first group, young males received a drinking water containing 30 mg of CdCl₂/L, for 90 days. Ten one-month-old males without Cd intoxication served as a control group. After 90 days of daily peroral exposure, body weight, femoral weight, femoral length, cortical bone thickness and histological structure of the femora were analysed.

Results

We found that subchronic peroral application of Cd had no significant effect on body weight, femoral length and cortical bone thickness in adult rats. On the other hand, femoral weight was significantly increased (P < 0.05) in Cd-intoxicated rats. These rats also displayed different microstructure in the middle part of the compact bone where vascular canals expanded into central area of substantia compacta and supplied primary and secondary osteons. Additionally, a few resorption lacunae which are connected with an early stage of osteoporosis were identified in these individuals. Histomorphometrical evaluations showed that all variables (area, perimeter, maximum and minimum diameter) of the primary osteons’ vascular canals, Haversian canals and secondary osteons were significantly decreased (P < 0.05) in the Cd group rats. This fact points to alterations in bone vascularization.

Conclusions

Subchronic peroral exposure to Cd significantly influences femoral weight and histological structure of compact bone in adult male rats. It induces an early stage of osteoporosis and causes reduced bone vascularization. Histomorphometrical changes of primary and secondary osteons allow for the conclusion that the bone mechanical properties could be weakened in the Cd group rats. The current study significantly expands the knowledge on damaging action of Cd on the bone.     

Negligible contribution from roots to soil-borne phospholipid fatty acid fungal biomarkers 18:2ω6,9 and 18:1ω9

The phospholipid fatty acid biomarkers 18:1ω9, 18:2ω6,9 and 18:3ω3,6,9 are commonly used as fungal biomarkers in soils. They have, however, also been found to occur in plant tissues, such as roots. Thus, the use of these PLFAs as fungal biomarkers in sieved soil, which may still contain small remains of roots, has been questioned. We used data from a recent beech tree girdling experiment to calculate the contribution of roots to these biomarkers and were able to demonstrate that not more than 0.61% of 18:1ω9 and 18:2ω6,9 in sieved soil samples originated from roots (but 4% of 18:3ω3,6,9). Additionally, the abundance of the biomarker 18:2ω6,9 in the soil was found to be highly correlated to ectomycorrhizal root colonization, which further corroborates its fungal origin. PLFA biomarkers were substantially reduced in vital roots from girdled trees compared to roots of control trees (by up to 76%), indicating that the major part of PLFAs measured in roots may actually originate from ectomycorrhizal fungi growing inside the roots. We calculated, that even a near to 50% reduction in fine root biomass - as observed in the girdling treatment - accounted for only 0.8% of the measured decrease of 18:2ω6,9. Our results demonstrate that both 18:1ω9 and 18:2ω6,9 are suitable biomarkers for detecting fungal dynamics in soils and that especially 18:2ω6,9 is a reliable biomarker to study mycorrhizal dynamics in beech forests.