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.     

Cyanidadsorption an Sesquioxiden,Tonmineralen und Huminstoffen

Cyanide adsorption on sesquioxides, clay-minerals and humic substances The adsorption of cyanide (KCN) on sesquioxides, clay minerals, and humic substances at different pH-values was studied. Moreover we looked for the CN-adsorption on L-layers of the humus forms mull, moder and mor. Cyanide was only adsorbed by humic acid. The amount of CN adsorbed increased with increasing pH of the reaction solution. IR-spectroscopic investigations of CN treated humic acids revealed that the cyanide was adsorbed at low pH (<7) as HCN-molecules by formation of hydrogen bonds with COOH-, COH-, OH- and NH₂-groups of the humic acid. At pH > 7 the cyanide was mainly adsorbed as CN^? by charge transfer with acceptor-molecules such as chinones. The cyanide adsorption of L-layers of humus forms decreased in the order mor > mull > moder. It is surmised that the HCN-molecules were not only adsorbed by humic acids in these layers but also by oxidation products of lignin, pectin, protein, cellulose, and carbon-hydrates of fulvic acids. Solutions of K₂HPO₄ did not desorb cyanides from humic acids to any great extent.     

Determination of oleandrin in tissues and biological fluids by liquid chromatography-electrospray tandem mass spectrometry

A rapid LC-MS/MS method, using a triple-quadrupole/linear ion trap mass spectrometer, was developed for the quantitative determination of oleandrin in serum, urine, and tissue samples. Oleandrin, the major cardiac glycoside of oleander (Nerium oleander L.), was extracted from serum and urine samples with methylene chloride and from tissues with acetonitrile. The tissue extracts were cleaned up using Florisil solid-phase extraction columns. Six replicate fortifications of serum and urine at 0.001 microg/g (1 ppb) oleandrin gave average recoveries of 97% with 5% CV (relative standard deviation) and 107% with 7% CV, respectively. Six replicate fortifications of liver at 0.005 microg/g (5 ppb) oleandrin gave average recoveries of 98% with 6% CV. This is the first report of a positive mass spectrometric identification and quantitation of oleandrin in tissue samples from oleander intoxication cases. The sensitivity and specificity of the LC-MS/MS analysis enables it to be the method of choice for toxicological investigations of oleander poisoning.     

Diagnosis and approach to poisoning in the horse.

Poisoning in the horse can present a highly complex case. The practitioner, owner, toxicologist, and pathologist play important roles, and all contribute information that may be important to the case. Once all the information is available, all the evidence is collected (historical, clinical, pathologic, and analytic), and proper sampling of specimens has occurred, a complete summary of the findings can be provided to the client. Based on identification of a potential toxic source and, ultimately, the diagnosis, specific treatment of affected animals and prevention of additional cases can be initiated. Consultation with a veterinary toxicologist aids in the follow-up of a poisoning case and can help to provide a thorough background to assist in preventing reoccurrence.     

Measurement of temperature generation in the corium of bovine claws during claw trimming with an angle grinder using different grinding and cutting disks

On 80 bovine hindclaws from slaughtered cows a correct functional claw trimming (sole horn thickness 5 mm; group 1) and on further 80 hindclaws a standardised, incorrect claw trimming (sole horn thickness 2 mm; group 2) was performed using 4 different disks (one cutting, three grinding disks). Two thermocouples were inserted into the solar corium and the heat production on the corium was measured during claw trimming. During correct functional claw trimming with each of the 4 disks a temperature increase in the corium with a median of < or = 0.3 degrees C was calculated. During incorrect claw trimming (2 mm sole horn thickness) a temperature increase with a median of < or = 0.3 degrees C for the first three disks was determined. During claw trimming using the abrasive semiflexible plastic disk a median of 1.3 degrees C temperature increase was calculated. Comparison of the temperatures measured in the claws from group 1 and 2 showed a statistically significant difference between all four disks. Comparison of the temperature increase within each group (group 1 and 2) revealed a statistically significant difference between the cutting disk with steel blades and the abrasive, semiflexible plastic disk in both groups. The highest temperatures measured were 41.5 degrees C in group 1 using the disk with 70% amount of fitted hard metal granulate, and 42.2 degrees C in group 2 (temperature increase of 10.6 degrees C) using the abrasive semiflexible plastic disk. Under the experimental conditions of this study, only a very slight thermogenesis was found when a functional and correct claw trimming using the 4 tested disks was performed. This slight thermogenesis should not be able to cause thermic injury of the corium. By trimming the sole horn incorrectly (2 mm) the measured heat generation in the corium was minor, without danger of thermic insult, excepted for using the disk with a 70% amount of fitted hard metal granulate and the abrasive semiflexible plastic disk. Even, it is unlikely that a short term temperature increase of 10.6 degrees C can cause thermic insults of the solar corium.     

Diagnosis of zinc phosphide poisoning in chickens using a new analytical approach

Approximately 200 chickens were found dead after the flooring of a slat-and-litter house was breached. No clinical signs of illness were observed in the surviving birds. During necropsy, rolled oats were found in the chickens' crops and gizzards, and the contents had a petroleum-like odor. Histopathologic examination revealed severe pulmonary edema and congestion of the chickens' lungs, hearts, livers, and kidneys. Based on the history and necropsy findings, zinc phosphide exposure was suspected. Diagnosis of zinc phosphide poisoning has previously been based on history of exposure, identification of the bait material in the gastrointestinal tract, and chemical detection of phosphine gas. However, currently available diagnostic methods are nonconfirmatory, and may produce false positive results. The objective of this case report was to determine whether the sudden death described in these chickens was caused by the ingestion of zinc phosphide, by developing a sensitive and highly specific gas chromatography/mass spectrometry (GC/MS) methodology for analysis of the gastrointestinal samples submitted to the laboratory. It was also found that the determination of zinc concentrations in liver or kidney tissue or stomach contents is not a reliable indicator of zinc phosphide poisoning.     

Identification of transgenes from wood of genetically transformed poplar trees

We have extracted total DNA from different fractions of fresh wood as well as from cold-stored and air-dried wood harvested from transgenic aspen grown in the field. The highest amounts of DNA were obtained from bark/cambium tissue; the DNA quality, however, was poor. Best results in PCR and Southern blot analyses were obtained from DNA extracted from early wood. Using appropriate primer pairs, amplification products were obtained from both the foreign gene (transgene) and aspen genomic sequences. In Southern blot analyses transgene-specific hybridisation signals were obtained. This is the first report on the detection of foreign genes in wood sampled from genetically modified trees.     

The occurrence of transmissible spongiform encephalopathies in roe deer (Capreolus capreolus), red deer (Cervus elaphus) and chamois (Rupicapra rupicapra) in Bavaria

Brain samples of 849 free-ranging ruminants (654 roe deer, 189 red deer and 6 chamois) from Bavaria were examined for TSE. In this respect, detections of PrPres were carried out by ELISA technique (BioRad) and immunohistochemistry using the monoclonal antibody (mAb) L42. Validation experiments of the ELISA technique for the detection of prionprotein from free-ranging ruminants showed that the examination of brains of wild ruminants lead to lower extinctions than that of bovine brain samples. Due to the fact that brain samples of 2 known CWD positive wapitis showed definite positive results, all samples were analysed by this test. There were no indications on the occurrence of TSE in any of the sample. In addition, immunohistochemical investigations of 10% of all samples did not show PrPres. Due to the large number of samples, the prevalence of TSE in Bavaria can be quoted under 0.5% for roe deer and under 1.5% for red deer with a certainty of 95%.