Fusarium mycotoxins: overlooked aquatic micropollutants?

Deoxynivalenol and zearalenone are among the most prevalent toxins produced by Fusarium spp. They have been investigated in food and feed products for decades but rarely in the environment. We therefore established solid-phase extraction and liquid chromatography-mass spectrometry (LC-MS) methods to quantify these mycotoxins at trace concentrations in aqueous natural samples. In a model emission study, we inoculated a winter wheat field with Fusarium graminearum and subsequently monitored deoxynivalenol and zearalenone in its drainage water. Before during and after harvest in June and July 2007, these toxins were emitted in concentrations from 23 ng/L to 4.9 microg/L for deoxynivalenol and from not detected to 35 ng/L for zearalenone. Simultaneously, in July and August 2007, deoxynivalenol was also detected in a number of Swiss rivers in concentrations up to 22 ng/L and zearalenone was present in several river samples below the method quantification limit. Other mycotoxins might be emitted from Fusarium-infected fields as well, because some of them are produced in similar amounts as deoxynivalenol and zearalenone and exhibit similar or even higher water solubility than deoxynivalenol. The ecotoxicological consequences of the presence of mycotoxins in surface waters remain to be elucidated.     

Hyperphosphorylation of cereal starch

Plant starch is naturally phosphorylated at a fraction of the C6 and the C3 hydroxyl groups during its biosynthesis in plastids. Starch phosphate esters are important in starch metabolism and they also generate specific industrial functionality. Cereal grains starch contains little starch bound phosphate compared with potato tuber starch and in order to investigate the effect of increased endosperm starch phosphate, the potato starch phosphorylating enzyme glucan water dikinase (StGWD) was overexpressed specifically in the developing barley endosperm. StGWD overexpressors showed wild-type phenotype. Transgenic cereal grains synthesized starch with higher starch bound phosphate content (7.5 (±0.67) nmol/mg) compared to control lines (0.8 (±0.05) nmol/mg) with starch granules showing altered morphology and lower melting enthalpy. Our data indicate specific action of GWD during starch biosynthesis and demonstrates the possibility for in planta production of highly phosphorylated cereal starch.     

Fundamental study on protein changes taking place during malting of oats

During the malting process, storage proteins are degraded by proteolytic enzymes into small peptides and amino acids. The activity of these enzymes was measured during malting of oats and was found to be increased. To quantify proteolytic degradation, proteins of unmalted, germinating and malted grains were fractionated. After extracting the oat proteins (Osborne fractionation), protein fractions were analysed using a Lab-on-a-Chip technique, which separates the proteins – based on their molecular weight – by capillary electrophoresis. This new technique for the analysis of proteins was supported by using two-dimensional gel electrophoresis. In addition, amino acid analysis was carried out. In general a degradation of proteins to small peptides and amino acids could be observed in the globulin, prolamin and glutelin fractions. In the albumin fraction a protein increase was observed, which is due to the fact that this fraction contains the majority of the metabolically active proteins. Amino acid analysis supported the observation of increased protein amount in the albumin fraction and decreased protein amounts in the other fractions. Some proteins, which have not been described in the literature, were detected in the albumin and glutelin fraction, since Lab-on-a-Chip technique allows detection of proteins with low molecular weights of 4.5 kDa.     

Soil water storage appears to compensate for climatic aridity at the xeric margin of European tree species distribution

Based on macroecological data, we test the hypothesis whether European tree species of temperate and boreal distribution maintain their water and nutrient supply in the more arid southern margin of their distribution range by shifting to more fertile soils with higher water storage than in their humid core distribution range (cf. soil compensatory effects). To answer this question, we gathered a large dataset with more than 200,000 plots that we related to summer aridity (SA), derived from WorldClim data, as well as soil available water capacity (AWC) and soil nutrient status, derived from the European soil database. The soil compensatory effects on tree species distribution were tested through generalized additive models. The hypothesis of soil compensatory effects on tree species distribution under limiting aridity was supported in terms of statistical significance and plausibility. Compared to a bioclimatic baseline model, inclusion of soil variables systematically improved the models’ goodness of fit. However, the relevance measured as the gain in predictive performance was small, with largest improvements for P. sylvestris, Q. petraea and A. alba. All studied species, except P. sylvestris, preferred high AWC under high SA. For F. sylvatica, P. abies and Q. petraea, the compensatory effect of soil AWC under high SA was even more pronounced on acidic soils. Soil compensatory effects might have decisive implications for tree species redistribution and forest management strategies under anthropogenic climate change. Therefore, soil compensatory effects deserve more intensive investigation, ideally, in studies combining different spatial scales to reduce the uncertainty associated with the precision of soil information.     

Molecular cloning and characterization of markers and cytokines for equid myeloid cells

The myeloid cell system comprises of monocytes, macrophages (MPhi), dendritic cells (DC), Kupffer cells, osteoclasts or microglia and is also known as the mononuclear phagocytic system (MPS). Essential cytokines to differentiate or activate these cells include GM-CSF or IL-4. Important markers for characterization include CD1, CD14, CD68, CD163 and CD206. All these markers, however, were not cloned or further characterized in equids by use of monoclonal antibodies earlier. To overcome this problem with the present study, two approaches were used. First, we cloned equine cytokines and markers, and second we analyzed cross-reactivity of human homologues or anti-human monoclonal antibodies. For cloning of equine cytokines and markers, we used degenerate primers delineated from other species, or equine-specific primers based on previous information in Genbank. Flow cytometry was used to determine the expression of markers on myeloid cells. Cross-reactivity could be shown for anti-human CD14, CD163 and mannose receptor (CD206) mAbs. Surface markers such as CD1 and CD68 that distinguish MPhi and DC were cloned and sequenced. According to blast homology, equine CD1a and CD1b could be identified and distinguished. With the resulting information, dendritic cells and macrophages of horses may be characterized.     

Investigation of the prevalence of summer eczema in Icelandic horses in northern Germany

An epidemiological study on summer eczema (SE) was performed in 490 Icelandic horses from 24 stud farms located in Lower Saxony and Westphalia. The study revealed a prevalence of summer eczema of 29.8 %. Horses imported from Iceland were affected with a frequency of 71.9 %, whereas horses born in Germany and other countries had a prevalence of 15.6 % and 27.3 %. Mares (33.1 %) and geldings (29.1 %) were more often affected by summer eczema than stallions with 15.5 %. The most common sites of summer eczema were mane and tail. Five to eight years old horses were more severely affected by SE than younger or older horses. Piebald horses showed higher degrees of SE than grey horses. The high prevalence of SE in Icelandic horses kept in Germany was due to imported horses from Iceland particularly horses imported at an age of seven to 15 years were at an extremely high risk to SE (96.4 %). The prevalence of SE significantly increased with higher age. Chestnut and grey horses born in Germany were prone to a lower risk to SE than bay, black or piebald horses. In horses imported from Iceland, a higher inbreeding coefficient was significantly related to a lower prevalence of SE. The most efficient measure recommended for reduction of SE in Germany would be to abandon imports of Icelandic horse from Iceland.     

Differentiation of five strains of infectious bursal disease virus: development of a strain-specific multiplex PCR

Infectious bursal disease virus (IBDV) is a major cause of disease problems in the poultry industry and vaccination has therefore been applied intensively to control the infection. The classical methods of detection and characterization of IBDV are by the use of immunodiffusion test and histopathology. Since these methods are laborious and have low specificity alternatives are needed. In the present study, we report the development of a strain-specific multiplex RT-PCR technique, which can detect and differentiate between field strains of IBDV and vaccine virus strains including a so-called hot vaccine strain widely used in the European poultry industry. The method, which is highly specific, fast and inexpensive, can be applied in all laboratories with basal PCR capabilities and equipment.     

Comparison of ultrasonography, computed tomography, and single-photon emission computed tomography for the detection and localization of canine insulinoma

Accurate preoperative detection, localization, and staging of the primary tumor and metastases are essential for the selection of appropriate candidates for surgery. In dogs with insulinoma, preoperative assessment usually is performed with transabdominal ultrasonography (US). There are no reports on the use of computed tomography (CT) for this purpose. The preoperative use of somatostatin receptor scintigraphy (SRS) recently has been advocated for the identification of insulinoma and gastrinoma in dogs, but its accuracy remains to be established. In this report US, CT, and single-photon emission computed tomography (SPECT) with [111In-DTPA-D-Phe1]-octreotide (a specific form of SRS) were compared for their effectiveness in detecting and localizing primary and metastatic insulinoma in dogs. Findings at surgery or postmortem examination served as control. Of 14 primary insulinomas, 5, 10, and 6 were correctly identified by US, CT, and SPECT, respectively. No lymph node metastases were detected by US or SPECT. CT identified 2 of 5 lymph node metastases but also identified 28 false-positive lesions. Two of 4 livers were found to be positive for metastases by 1 of the imaging techniques. US can be used for the initial evaluation of dogs with hypoglycemia. Although CT identifies most primary tumors, intraoperative inspection and palpation of the pancreas is still superior. SPECT appears as effective as US and CT in detecting insulinomas. Future developments in preoperative imaging techniques might improve current methods of canine insulinoma detection.