On-farm herbicide use in winter wheat and winter oilseed rape in Mecklenburg-Vorpommern as influenced by crop rotation

Aus einem umfangreichen Datensatz landwirtschaftlicher Praxis-Daten wurden feldspezifische Fruchtsequenzen über einen Zeitraum von neun Jahren ausgewertet. Das Ziel der vorliegenden Studie ist es, den kombinierten Einfluss der Fruchtfolgegestaltung und Bodenbearbeitung auf den Herbizid-Einsatz landwirtschaftlicher Betriebe in Mecklenburg-Vorpommern zu prüfen. Weiterhin wird der Zusammenhang zwischen Fruchtfolge, Bodenbearbeitung, Herbizidkosten und dem Ertrag untersucht. Dazu werden dreifeldrige Fruchtsequenzen bezüglich ihrer Anfälligkeit gegenüber Unkräutern in Risikoklassen eingestuft und damit die indirekten Risiken der Vorfrüchte betrachtet. Winterweizen und Winterraps werden in Mecklenburg-Vorpommern überwiegend in Fruchtsequenzen mit hohem Risiko gegenüber dem Auftreten von Unkräutern und Ungräsern angebaut. Es zeigt sich, je diverser die Fruchtsequenzgestaltung, bei gleichzeitiger Intensivierung der Bodenbearbeitung, desto geringer ist die Intensität der eingesetzten Herbizide sowie der monetäre Aufwand für die Unkrautbekämpfung in beiden Kulturen. In Fruchtsequenzen mit hohem Anteil an Sommerkulturen und unter wendender Bodenbearbeitung wurde signifikant mehr Weizen geerntet. Fruchtfolge und Bodenbearbeitung sind bedeutende Maßnahmen im integrierten Unkrautmanagement. Die Umsetzung in der Praxis erfordert Unterstützung der Landwirte durch Forschung, Beratungsdienste und Politik.Nine-year data on chemical plant protection measures and field management were examined in winter wheat and winter oilseed rape. We investigated the contribution of crop rotation to the variability of herbicide use of farms in the north German region Mecklenburg-Vorpommern. Our study aimed at answering the question: do diverse crop sequences and intensive tillage practices lead to reduced herbicide use and herbicide costs in arable farming. Furthermore, we analyzed the field-specific relation between crop rotation and yield. We classified crop sequences (triplets of three succeeding crops) according to their susceptibility for weeds (= 'riskiness'). The crop sequences in our analysis were mostly classified as highly risky for weed infestations in winter wheat and winter oilseed rape. Herbicide use intensities and costs were smaller in more diverse crop sequences. Crop sequences including a high proportion of summer crops decreased herbicide use in winter wheat and winter oilseed rape. Moreover, our results prove that the crop diversification together with ploughing is an effective management measure, which leads to higher yields in winter wheat. Crop rotation and tillage practices are important methods to control weeds in integrated weed management. More efforts on an advisory, scientific and political level are necessary to inform farmers in more detail about the advantages of crop diversity

Analysing Farmers’ Herbicide Use Pattern to Estimate the Magnitude and Field-Economic Value of Crop Diversification

We present an on-farm approach to measure the effect of crop diversification on farmers’ field economic values. Eleven years of data (2010–2020) on the chemical herbicide use, tillage practices and crop yields of 17 farms in north-eastern Germany were examined for winter wheat (WW) and winter oilseed rape (WOSR). We used a common conceptual framework to classify farmers’ crop sequences according to their susceptibility to weeds (‘riskiness’). Linear mixed models were used to analyse the relationship between crop sequence, tillage practice (inversion/non-inversion) and the response variables ‘total herbicide costs’, ‘crop yield’ and ‘economic income’. Our results indicate that farmers in the area surveyed commonly grow crop sequences with a high risk of weeds. The driving forces behind this classification are high ratios of winter cereals and WOSR in the sequences. The most interesting result of our analysis is that farmers’ total herbicide costs (THCfy) significantly decreased from a higher to a lower riskiness class. Diversified crop sequences decreased the THCfy for WW by up to 12 EUR ha−1 and for WOSR by 19–56 EUR ha−1. Considering the crop diversification effects, the combined influence of tillage and crop sequence seems to be important. Significant differences in crop yield between the riskiness classes were found in WW and WOSR solely in non-inversion tillage systems. Hence, the analysis of farmers’ ‘economic income’ revealed the great impact of crop diversification for non-inversion tillage systems. Indeed, we found that simplifying both crop sequence diversity and tillage intensity implies higher herbicide costs and, thereby, higher economic input. The best strategy for reducing herbicide costs in WW and WOSR cropping is to increase the use of summer crops or field grass as previous crops

Major Determinants of Airway Epithelial Cell Sensitivity to S. aureus Alpha-Toxin: Disposal of Toxin Heptamers by Extracellular Vesicle Formation and Lysosomal Degradation

Alpha-toxin is a major virulence factor of Staphylococcus aureus. Monomer binding to host cell membranes results in the formation of heptameric transmembrane pores. Among human model airway epithelial cell lines, A549 cells were most sensitive toward the toxin followed by 16HBE14o- and S9 cells. In this study we investigated the processes of internalization of pore-containing plasma membrane areas as well as potential pathways for heptamer degradation (lysosomal, proteasomal) or disposal (formation of exosomes/micro-vesicles). The abundance of toxin heptamers upon applying an alpha-toxin pulse to the cells declined both in extracts of whole cells and of cellular membranes of S9 cells, but not in those of 16HBE14o- or A549 cells. Comparisons of heptamer degradation rates under inhibition of lysosomal or proteasomal degradation revealed that an important route of heptamer degradation, at least in S9 cells, seems to be the lysosomal pathway, while proteasomal degradation appears to be irrelevant. Exosomes prepared from culture supernatants of toxin-exposed S9 cells contained alpha-toxin as well as low amounts of exosome and micro-vesicle markers. These results indicate that lysosomal degradation of internalized toxin heptamers may be the most important determinant of toxin-resistance of some types of airway epithelial cells

S. aureus alpha-toxin monomer binding and heptamer formation in host cell membranes – Do they determine sensitivity of airway epithelial cells toward the toxin?

Alpha-toxin (Hla) is a major virulence factor of Staphylococcus aureus (S. aureus) and plays an important role in S. aureus-induced pneumonia. It binds as a monomer to the cell surface of eukaryotic host cells and forms heptameric transmembrane pores. Sensitivities toward the toxin of various types of potential host cells have been shown to vary substantially, and the reasons for these differences are unclear. We used three human model airway epithelial cell lines (16HBE14o-, S9, A549) to correlate cell sensitivity (measured as rate of paracellular gap formation in the cell layers) with Hla monomer binding, presence of the potential Hla receptors ADAM10 or α5β1 integrin, presence of the toxin-stabilizing factor caveolin-1 as well as plasma membrane lipid composition (phosphatidylserine/choline, sphingomyelin). The abundance of ADAM10 correlated best with gap formation or cell sensitivities, respectively, when the three cell types were compared. Caveolin-1 or α5β1 integrin did not correlate with toxin sensitivity. The relative abundance of sphingomyelin in plasma membranes may also be used as a proxi for cellular sensitivity against alpha-toxin as sphingomyelin abundances correlated well with the intensities of alpha-toxin mediated gap formation in the cell layers

Sphingomyelin Depletion from Plasma Membranes of Human Airway Epithelial Cells Completely Abrogates the Deleterious Actions of <em>S. aureus</em> Alpha-Toxin

Interaction of Staphylococcus aureus alpha-toxin (hemolysin A, Hla) with eukaryotic cell membranes is mediated by proteinaceous receptors and certain lipid domains in host cell plasma membranes. Hla is secreted as a 33 kDa monomer that forms heptameric transmembrane pores whose action compromises maintenance of cell shape and epithelial tightness. It is not exactly known whether certain membrane lipid domains of host cells facilitate adhesion of Ha monomers, oligomerization, or pore formation. We used sphingomyelinase (hemolysin B, Hlb) expressed by some strains of staphylococci to pre-treat airway epithelial model cells in order to specifically decrease the sphingomyelin (SM) abundance in their plasma membranes. Such a pre-incubation exclusively removed SM from the plasma membrane lipid fraction. It abrogated the formation of heptamers and prevented the formation of functional transmembrane pores. Hla exposure of rHlb pre-treated cells did not result in increases in [Ca2+]i, did not induce any microscopically visible changes in cell shape or formation of paracellular gaps, and did not induce hypo-phosphorylation of the actin depolymerizing factor cofilin as usual. Removal of sphingomyelin from the plasma membranes of human airway epithelial cells completely abrogates the deleterious actions of Staphylococcus aureus alpha-toxin

ATP Release from Human Airway Epithelial Cells Exposed to Staphylococcus aureus Alpha-Toxin

Airway epithelial cells reduce cytosolic ATP content in response to treatment with S. aureus alpha-toxin (hemolysin A, Hla). This study was undertaken to investigate whether this is due to attenuated ATP generation or to release of ATP from the cytosol and extracellular ATP degradation by ecto-enzymes. Exposure of cells to rHla did result in mitochondrial calcium uptake and a moderate decline in mitochondrial membrane potential, indicating that ATP regeneration may have been attenuated. In addition, ATP may have left the cells through transmembrane pores formed by the toxin or through endogenous release channels (e.g., pannexins) activated by cellular stress imposed on the cells by toxin exposure. Exposure of cells to an alpha-toxin mutant (H35L), which attaches to the host cell membrane but does not form transmembrane pores, did not induce ATP release from the cells. The Hla-mediated ATP-release was completely blocked by IB201, a cyclodextrin-inhibitor of the alpha-toxin pore, but was not at all affected by inhibitors of pannexin channels. These results indicate that, while exposure of cells to rHla may somewhat reduce ATP production and cellular ATP content, a portion of the remaining ATP is released to the extracellular space and degraded by ecto-enzymes. The release of ATP from the cells may occur directly through the transmembrane pores formed by alpha-toxin

Staphylococcus aureus Alpha-Toxin in Deep Tracheal Aspirates—Preliminary Evidence for Its Presence in the Lungs of Sepsis Patients

The pore forming alpha-toxin (hemolysin A, Hla) of Staphylococcus aureus (S. aureus) is a major virulence factor with relevance for the pathogenicity of this bacterium, which is involved in many cases of pneumonia and sepsis in humans. Until now, the presence of Hla in the body fluids of potentially infected humans could only be shown indirectly, e.g., by the presence of antibodies against Hla in serum samples or by hemolysis testing on blood agar plates of bacterial culture supernatants of the clinical isolates. In addition, nothing was known about the concentrations of Hla actually reached in the body fluids of the infected hosts. Western blot analyses on 36 samples of deep tracheal aspirates (DTA) isolated from 22 hospitalized sepsis patients using primary antibodies against different epitopes of the Hla molecule resulted in the identification of six samples from five patients containing monomeric Hla (approx. 33 kDa). Two of these samples showed also signals at the molecular mass of heptameric Hla (232 kDa). Semiquantitative analyses of the samples revealed that the concentrations of monomeric Hla ranged from 16 to 3200 ng/mL. This is, to our knowledge, the first study directly showing the presence of S. aureus Hla in samples of airway surface liquid in human patients

Target Mechanisms of the Cyanotoxin Cylindrospermopsin in Immortalized Human Airway Epithelial Cells

Cylindrospermopsin (CYN) is a cyanobacterial toxin that occurs in aquatic environments worldwide. It is known for its delayed effects in animals and humans such as inhibition of protein synthesis or genotoxicity. The molecular targets and the cell physiological mechanisms of CYN, however, are not well studied. As inhalation of CYN-containing aerosols has been identified as a relevant route of CYN uptake, we analyzed the effects of CYN on protein expression in cultures of immortalized human bronchial epithelial cells (16HBE14o−) using a proteomic approach. Proteins whose expression levels were affected by CYN belonged to several functional clusters, mainly regulation of protein stability, cellular adhesion and integration in the extracellular matrix, cell proliferation, cell cycle regulation, and completion of cytokinesis. With a few exceptions of upregulated proteins (e.g., ITI inhibitor of serine endopeptidases and mRNA stabilizer PABPC1), CYN mediated the downregulation of many proteins. Among these, centrosomal protein 55 (CEP55) and osteonectin (SPARC) were significantly reduced in their abundance. Results of the detailed semi-quantitative Western blot analyses of SPARC, claudin-6, and CEP55 supported the findings from the proteomic study that epithelial cell adhesion, attenuation of cell proliferation, delayed completion of mitosis, as well as induction of genomic instability are major effects of CYN in eukaryotic cells