Report on the 42nd joint meeting of the DPG-working group Nematology and the working group “free living nematodes”

Journal of Plant Diseases and Protection -     

Primed N2O emission from native soil nitrogen: A 15N‐tracing laboratory experiment

Soils can naturally be a source of the potent greenhouse gas nitrous oxide (N₂O). By contrast, the largest anthropogenic source of N₂O is the application of nitrogen (N) fertilizer on agricultural soil, but it is unclear if fertilizer‐supported N₂O emission only originates from the fertilizer N directly or through additionally stimulated N₂O production from native soil N. Even though native soil N also includes mineral N already in soil before fertilizer application, organic N is the principal native N pool and thereby provides for mineral N cycling and N₂O emission. Here, we tested (1) the contribution of native soil N to N₂O emission after mineral N fertilizer application and (2) whether it is affected by different soil organic matter (SOM) contents by conducting a laboratory ¹⁵N‐tracing experiment with agricultural soil from a long‐term field trial with two treatments. Both field treatments are fertilized with mineral N, whereas only one of the two receives liquid manure causing higher SOM content. Soil sampling was conducted in March 2016 shortly before fertilizer application in the field. The application of ¹⁵N‐labeled fertilizer more than doubled the N₂O production from native N sources compared to the non‐fertilized control incubations. This primed N₂O production contributed by 5–8% to the fertilizer‐induced N₂O emission after one week of incubation and was similar for both field treatments regardless of liquid manure application. Therefore, further research is needed to link N₂O priming to its potential production pathways and sources. While the observed effect may be important in soils, the amount of applied N fertilizer remains the largest concern being responsible for the majority of N₂O emission.     

Effect of fungicides on the complex of <Emphasis Type="Italic">Fusarium</Emphasis> species and saprophytic fungi colonizing wheat kernels

Fusarium head blight of wheat, often associated with mycotoxin contamination of food and feed is caused by various Fusarium species. The efficacy of fungicide sprays for the control of the disease and mycotoxins varies from being highly effective to even increasing mycotoxin levels. The potential role of antagonistic fungi in this variability was investigated assessing sensitivity of Fusarium species and saprophytic fungi colonizing wheat kernels to fungicides. Saprophytes were tested for their antagonistic activity to the prevalent Fusarium species Fusarium avenaceum, Fusarium culmorum, Fusarium graminearum, and Fusarium poae. Fungal isolates from mature winter wheat kernels were Alternaria alternata, Arthrinium sp., Aspergillus niger, Epicoccum sp., Microdochium spp., Rhizopus oryzae and Trichoderma sp. In dual culture A. niger, R. oryzae and Trichoderma hamatum were more effective in reducing mycelial growth of Fusarium species than Microdochium majus; A. alternata and Epicoccum sp. were ineffective because of slow growth rates. Saprophytic fungi were sensitive to triazoles; however, prothioconazole and tebuconazole had stronger effects on mycelial growth of Fusarium species. ED₅₀ values also indicated significant differences in the sensitivity of Fusarium species to triazoles (range 0.1–1.7 mg l⁻¹). Azoxystrobin and fluoxastrobin were largely ineffective in inhibiting in vitro growth of Fusarium spp.; sensitivity of the other fungi was generally lower, except for M. majus which was highly sensitive. Due to differences in fungicide sensitivity among Fusarium spp. and ear-colonizing fungi antagonistic to Fusarium spp. fungicides are likely to modify the balance within the mycoflora of wheat ears which may also affect the mycotoxin contamination of grain.     

Determination of lignans in edible and nonedible parts of pomegranate (Punica granatum L.) and products derived therefrom, particularly focusing on the quantitation of isolariciresinol using HPLC-DAD-ESI/MSn

A method for the characterization and quantitation of phyto-estrogenic lignans from pomegranate (Punica granatum L.) fruits and fruit-derived products by HPLC-DAD-MS(n) was developed. For this purpose, edible and nonedible parts of pomegranate (aril, peel, mesocarp, seed, and twigs), commercial juices, juices produced on pilot-plant scale, and encapsulated dietary supplements were analyzed. In addition to the peel, mesocarp, and twigs, lignans were detected in two juices obtained from entire fruits, four commercial juices, and three encapsulated pomegranate extracts. Isolariciresinol was the predominant lignan with contents of 5.0, 10.5, and 45.8 mg/kg dry matter in processed pomegranate mesocarp, peel, and twigs, respectively. In contrast, due to their low amounts, quantitation of lignans in pomegranate products was impossible. Therefore, contrary to previous assumptions, lignans were found to be less relevant in pomegranate-derived products. However, the byproduct from pomegranate processing may be used for lignan extraction. The method presented allows one to differentiate between pomegranate-derived products obtained from fruits without peels or by dejuicing applying low pressures, which were devoid of lignans, and those obtained from entire fruits applying high pressures, thus containing lignans. Consequently, this study helps to optimize process technology aiming at the recovery of preparations with well-desired compositions, which may reduce the risk of a wide range of diseases, such as certain types of cancer.     

The genome of the African trypanosome Trypanosoma brucei

African trypanosomes cause human sleeping sickness and livestock trypanosomiasis in sub-Saharan Africa. We present the sequence and analysis of the 11 megabase-sized chromosomes of Trypanosoma brucei. The 26-megabase genome contains 9068 predicted genes, including approximately 900 pseudogenes and approximately 1700 T. brucei-specific genes. Large subtelomeric arrays contain an archive of 806 variant surface glycoprotein (VSG) genes used by the parasite to evade the mammalian immune system. Most VSG genes are pseudogenes, which may be used to generate expressed mosaic genes by ectopic recombination. Comparisons of the cytoskeleton and endocytic trafficking systems with those of humans and other eukaryotic organisms reveal major differences. A comparison of metabolic pathways encoded by the genomes of T. brucei, T. cruzi, and Leishmania major reveals the least overall metabolic capability in T. brucei and the greatest in L. major. Horizontal transfer of genes of bacterial origin has contributed to some of the metabolic differences in these parasites, and a number of novel potential drug targets have been identified.     

Metabolism and persistence of atrazine in several field soils with different atrazine application histories

To assess the potential occurrence of accelerated herbicide degradation in soils, the mineralization and persistence of (14)C-labeled and nonlabeled atrazine was evaluated over 3 months in two soils from Belgium (BS, atrazine-treated 1973-2008; BC, nontreated) and two soils from Germany (CK, atrazine-treated 1986-1989; CM, nontreated). Prior to the experiment, accelerated solvent extraction of bulk field soils revealed atrazine (8.3 and 15.2 μg kg(-1)) in BS and CK soils and a number of metabolites directly after field sampling, even in BC and CM soils without previous atrazine treatment, by means of LC-MS/MS analyses. For atrazine degradation studies, all soils were incubated under different moisture conditions (50% maximum soil water-holding capacity (WHC(max))/slurried conditions). At the end of the incubation, the (14)C-atrazine mineralization was high in BS soil (81 and 83%) and also unexpectedly high in BC soil (40 and 81%), at 50% WHC(max) and slurried conditions, respectively. In CK soil, the (14)C-atrazine mineralization was higher (10 and 6%) than in CM soil (4.7 and 2.7%), but was not stimulated by slurried conditions. The results revealed that atrazine application history dramatically influences its degradation and mineralization. For the incubation period, the amount of extractable atrazine, composed of residues from freshly applied atrazine and residues from former field applications, remained significantly greater (statistical significance = 99.5 and 99.95%) for BS and CK soils, respectively, than the amount of extractable atrazine in the bulk field soils. This suggests that (i) mostly freshly applied atrazine is accessible for a complex microbial community, (ii) the applied atrazine is not completely mineralized and remains extractable even in adapted soils, and (iii) the microbial atrazine-mineralizing capacity strongly depends on atrazine application history and appears to be conserved on long time scales after the last application.     

Influence of aging and salting on protein secondary structures and water distribution in uncooked and cooked pork. A combined FT-IR microspectroscopy and 1H NMR relaxometry study

Fourier transform infrared (FT-IR) microspectroscopy and low-field (LF) proton NMR transverse relaxation measurements were used to study the changes in protein secondary structure and water distribution as a consequence of aging (1 day and 14 days) followed by salting (3%, 6%, and 9% NaCl) and cooking (65 degrees C). An enhanced water uptake and increased proton NMR relaxation times after salting were observed in aged meat (14 days) compared with nonaged meat (1 day). FT-IR bands revealed that salting induced an increase in native beta-sheet structure while aging triggered an increase in native alpha-helical structure before cooking, which could explain the effects of aging and salting on water distribution and water uptake. Moreover, the decrease in T2 relaxation times and loss of water upon cooking were attributed to an increase in aggregated beta-sheet structures and a simultaneous decrease in native protein structures. Finally, aging increased the cooking loss and subsequently decreased the final yield, which corresponded to a further decrease in T2 relaxation times in aged meat upon cooking. However, salting weakened the effect of aging on the final yield, which is consistent with the increased T2 relaxation times upon salting for aged meat after cooking and the weaker effect of aging on protein secondary structural changes for samples treated with high salt concentration. The present study reveals that changes in water distribution during aging, salting, and cooking are not only due to the accepted causal connection, i.e., proteolytic degradation of myofibrillar structures, change in electrostatic repulsion, and dissolution and denaturation of proteins, but also dynamic changes in specific protein secondary structures.     

A comparison of cardiorespiratory variables during isoflurane-fentanyl and propofol-fentanyl anaesthesia for surgery in injured cats

OBJECTIVE: To compare haemodynamic and respiratory variables during isoflurane-fentanyl (IF) and propofol-fentanyl (PF) anaesthesia for surgery in injured cats. STUDY DESIGN: Prospective, randomized, controlled clinical study. ANIMALS: Thirty-three client-owned injured cats undergoing orthopaedic surgery. MATERIALS AND METHODS: Pre-anaesthetic medication was intravenous midazolam 1 mg kg(-1), butorphanol 0.4 mg kg(-1) and ketamine 2 mg kg(-1). Anaesthesia was induced with propofol (P) and maintained with either: (a) a continuous rate infusion (CRI) of fentanyl (F) 0.02 mg kg(-1) hour(-1) and isoflurane (initial end-tidal concentration of 1%), (b) a fentanyl CRI (dose as before) and sevoflurane (initial end-tidal concentration of 2%) or (c) a CRI of propofol (12 mg kg(-1) hour(-1)). All three techniques were given to effect until surgical anaesthesia was achieved. Heart rate and rhythm (ECG), mean arterial blood pressure, respiratory rate, tidal volume and end-tidal CO(2) concentration were recorded. Venous blood gas analysis was performed before and after sedation, and at the end of anaesthesia. Blood chemistry and blood cell counts were assessed before, at the end of, and 24 hours after anaesthesia. The variables recorded from cats anaesthetized with IF and PF were compared. RESULTS: Mean end-expiratory isoflurane concentration was 1.19 +/- 0.19%. The propofol infusion rate was 11.4 +/- 0.8 mg kg(-1) hour(-1). No significant differences between the two groups in heart rate were identified; no cardiac dysrhythmias were recorded. Mean arterial blood pressure was significantly lower in IF cats during skin incision (p = 0.01), during surgery without intense surgical stimulation (p < 0.01) and during surgery with intense surgical stimulation (p = 0.01). Nine of 11 cats in the IF group were markedly hypotensive (34-49 mmHg) while seven of 11 cats in group PF were mildly hypotensive (49-59 mmHg). One of 11 cats in group IF and nine of 11 cats in group PF required intermittent positive pressure ventilation (IPPV) to maintain end-tidal CO(2) levels below 6.66 kPa (50 mmHg). CONCLUSION AND CLINICAL RELEVANCE: Despite the necessity to ventilate the lungs of cats in the PF group, arterial blood pressure was better maintained. Propofol-fentanyl anaesthesia is better for surgery in injured cats providing the means to impose IPPV are available.     

Regional ventilation distribution and dead space in anaesthetized horses treated with and without continuous positive airway pressure: novel insights by electrical impedance tomography and volumetric capnography

Objective

The aim of this study was to evaluate the effect of continuous positive airway pressure (CPAP) on regional distribution of ventilation and dead space in anaesthetized horses.

Immunostimulation of bronchoalveolar lavage cells from recurrent airway obstruction-affected horses by different CpG-classes bound to gelatin nanoparticles

Recurrent airway obstruction (RAO) in horses has become a common problem in stabled horses in industrialized countries and deserves new therapeutic strategies. CpG-oligodeoxynucleotides (CpG-ODNs) were developed as effective immunostimulating agents to induce a Th2/Th1 shift. These agents showed a beneficial therapeutic effect in allergic diseases with predominant Th2 immunoresponse. CpG-ODN delivery by gelatin nanoparticles (GNPs) resulted in enhanced cellular uptake in murine and human in vitro studies and was a starting point for the present trial. The aim of this study was to identify an optimal stimulating CpG motif in horses with regard to species specificity on equine bronchoalveolar lavage (BAL) cells, in terms of a possible specific immunomodulation effect (Th2/Th1 shift) by used CpG-ODN. Accordingly, GNPs were evaluated as a delivery system to improve CpG-ODN immunostimulation in equine BAL cells. BAL fluid (BALF) was obtained from seven horses with moderate RAO and from four healthy horses and was subsequently incubated with five different CpG-ODN sequences (from A-, B- and C-class) and one ODN without any CpG motif. Release of three key cytokines (IL-4, IL-10 and IFN-γ) was quantified by ELISA to detect an allergy mediated Th2 immunoresponse (IL-4) as well as a proinflammatory Th1 response (IFN-γ). Due to its specific anti-inflammatory and anti-allergic effects, IL-10 was considered as a beneficial agent in pathophysiology of RAO. Results showed a significant upregulation of IL-10 and IFN-γ on the one hand and a downregulation of IL-4 on the other hand in RAO affected horses. Cell cultures from healthy horses had a significantly stronger response in cytokine release to all the applied stimuli in contrast to RAO derived cells. Comparing all five CpG sequences, A-class 2216 significantly showed the highest immunomodulatory effects on equine BALF cells and, hence, was chosen for follow-up preliminary clinical studies.