Mode of resistance to respiration inhibitors at the cytochrome bc1 enzyme complex of Mycosphaerella fijiensis field isolates

Field isolates of Mycosphaerella fijiensis, causing black Sigatoka of banana, were characterised for their sensitivity to different inhibitors of the cytochrome bc₁ enzyme complex (Qo respiration inhibitors, strobilurin fungicides), using physiological, biochemical and molecular genetic methods. Strobilurin‐resistant isolates exhibited very high resistance factors both in mycelial growth inhibition and NADH consumption assays. Cross‐resistance was observed among all Qo inhibitors, including trifloxystrobin, azoxystrobin, famoxadone, strobilurin B and myxothiazol. However, the Qi and the cytochrome aa₃ inhibitors, antimycin A and potassium cyanide, respectively, were not cross‐resistant to Qo inhibitors. In sensitive but not in resistant isolates, mixtures of Qo inhibitors and SHAM, an inhibitor of the alternative oxidase (AOX), were more active than the components alone, indicating that the alternative pathway is essential in metabolism, but not causal for resistance. In the cell‐free NADH‐consumption assay, the Qo inhibitors affected the sensitive but not the resistant isolates, suggesting that AOX was not active in sub‐mitochondrial particles. In whole cells, however, the AOX has a basic expression level and is probably not inducible by trifloxystrobin. Sequencing of the cytochrome b gene of sensitive and resistant M fijiensis isolates revealed a difference in the nucleotide sequence leading to a single amino acid change from glycine to alanine at position 143 in the resistant isolate. This change is known to occur also in the naturally tolerant basidiomycete Mycena galopoda. It is suggested that the field isolates of M fijiensis can acquire resistance to Qo inhibitors due to a target site alteration with a single base pair change. Resistant isolates do not seem to contain a mixture of mutated and non‐mutated DNA, indicating a complete selection of resistant mitochondria and a maternally donated mode of resistance. © 2000 Society of Chemical Industry     

Long-term trends of phosphorus nutrition and topsoil phosphorus stocks in unfertilized and fertilized Scots pine (Pinus sylvestris) stands at two sites in Southern Germany

For two Scots pine (Pinus sylvestris) ecosystems in S Germany with different atmospheric N deposition (Pfaffenwinkel, intermediate N deposition; Pustert, large N deposition), the supply with phosphorus (P) has been monitored for unfertilized and fertilized plots over more than four decades by foliar analysis (1964–2007). Additionally, topsoil concentrations and stocks of total P and plant-available P (citric-acid-extractable phosphate) were quantified in 10-year intervals (1982/1984, 1994, 2004). At both sites, fertilization experiments, including the variants control, NPKMgCa + lime, PKMgCa + lime + introduction of lupine, corresponding to an addition of 75 and 90 kg ha⁻¹ P in Pustert and Pfaffenwinkel, respectively had been established in 1964. Our study revealed different trends of the P nutritional status for the pines at the two sites during the recent four decades: At Pustert, elevated atmospheric N deposition together with small topsoil P pools resulted in significant deterioration of Scots pine P nutrition and in an increasingly unbalanced N/P nutrition. At Pfaffenwinkel a trend of improved P nutrition from 1964 to 1991 was replaced by an opposite trend in the most recent 15 years. For our study sites, which are characterized by acidic soils with thick O layers, the forest floor stock of citric-acid-extractable phosphate showed a strong and significant correlation with the P concentration in current-year pine foliage, and thus was an appropriate variable to predict the P nutritional status of the stands. Total P stocks as well as the concentrations of total P in the forest floor or in the mineral topsoil were poorly correlated with pine foliar P concentrations and thus inappropriate predictors of P nutrition. P fertilization in the 1960s sustainably improved the P nutritional status of the stands. At Pfaffenwinkel, foliar P concentrations and topsoil stocks of citric-acid-extractable phosphate were increased at the fertilized plots relative to the control plots even 40 years after fertilization; at Pustert, foliar P concentrations were increased for about 20 years.     

Examination of the prevalence of skin injuries in debeaked fattened turkeys

Within the framework of a cooperative research project, turkey health as well as numerous aspects of animal welfare were examined in various intensive commercial farms with varying rearing forms. The present study demonstrates the prevalence of skin injuries concerning living animals as well as carcasses of slaughtered turkeys of both sexes. Although all turkeys were debeaked, prevalences of skin injuries at a value of 12.8% with male turkeys and 13.8% with female turkeys could be found at the age of 16 weeks. Within the scope of medically inspections skin injuries were mainly established in the head region, especially the snood as well as in the region of the back. By means of their clinical picture pecking wounds and scratch injuries could be distinguished. Skin lesion produced primary as a result of scratching the skin surface, could be enlarged by the animal itself or by conspecifics by beak pecking, especially after bleeding, in spite of debeaked upper beak. Injuries in the head region, especially of the snood, could be explained mainly as a result of pecking by conspecifics by reason of their clinical picture and were found mainly in male turkeys. Skin injuries in the region of the back, especially at the coxal tuberosity, were identified primarily as scratch marks. They were found mainly in female turkeys. A statistically significant correlation of the injury prevalence to particular husbandry parameters discussed in the literature as "predisposing" or "limiting" factors (e.g., population density, light regime or offer of employment material) could not be established in this study. At the meat inspection fresh scratching injuries at the chest and the hind legs were diagnosed most often. Especially fresh injuries of the carcass point to a misguided behaviour of the staff concerning animal handling during loading and transport which is not conform to animal welfare.     

Toll-like receptor triggering of a vitamin D-mediated human antimicrobial response

In innate immune responses, activation of Toll-like receptors (TLRs) triggers direct antimicrobial activity against intracellular bacteria, which in murine, but not human, monocytes and macrophages is mediated principally by nitric oxide. We report here that TLR activation of human macrophages up-regulated expression of the vitamin D receptor and the vitamin D-1-hydroxylase genes, leading to induction of the antimicrobial peptide cathelicidin and killing of intracellular Mycobacterium tuberculosis. We also observed that sera from African-American individuals, known to have increased susceptibility to tuberculosis, had low 25-hydroxyvitamin D and were inefficient in supporting cathelicidin messenger RNA induction. These data support a link between TLRs and vitamin D-mediated innate immunity and suggest that differences in ability of human populations to produce vitamin D may contribute to susceptibility to microbial infection.     

Model Experiments on the Influence of Artificial Humic Compounds on Chemodenitrification

Chemodenitrification is of importance in both soils and the treatment of some types of wastewater. During model experiments,the impact of various conditions, such as pH and especially artificial humic matter and oxygen on this process was studied to build upkinetic models. The chemodenitrification rate decreased due to the ongoingautoxidation/polymerization of hydroquinone to artificial humic matterfrom 11.02 μg (L h)^-1 after 7 days autoxidation to 5.38 μg (L h)^-1 after 14 days at pH 4 under aerobic conditionsand an initial nitrite concentration of 250 μg L^-1. At the same pH,with the same nitrite concentration, and in the presence of Roth humic matter(2 mg L^-1) under aerobic conditions, the chemodenitrification rate was0.73 μg (L h)^-1, whereas under anaerobic conditions itwas considerably higher (2.88 μg (L h)^-1). In anothermodel experiment, it was shown that the amount of nitrite incorporated into the artificial humic matter was less then 1%. Further, it was found that the main reaction product of chemodenitrification is NO.     

Preparation and comparative release characteristics of three anthocyanin encapsulation systems

Bilberries (Vaccinium myrtillus L.) and their major polyphenolic constituents, anthocyanins, have preventive activities inter alia against colon cancer and inflammatory bowel diseases. However, anthocyanins are sensitive to environmental conditions; thus their bioavailability in the gastrointestinal tract is an important determinant of their in vivo activity. In the study reported here, the potential benefits of encapsulating an anthocyanin rich bilberry extract (BE) on anthocyanin stability were investigated. Nonencapsulated BE and three different BE loaded microcapsule systems were incubated in simulated gastric fluid (SGF) and fed state simulated intestinal fluid (FeSSIF). After exposure to these media, released anthocyanins were identified and quantified by HPLC with UV/Vis detection. Although a rapid release of anthocyanins was observed within the first 20 min, encapsulation of anthocyanins doubled the amount of available anthocyanins after 150 min of incubation. These results illustrate the ability of encapsulation to inhibit early degradation of anthocyanins in the intestinal system.     

Der Einfluß der natürlichen organischen Substanzen auf die Metallverteilung zwischen Boden und Bodenlösung in einem sauren Waldboden

The influence of organic matter in the translocation of metals between soil and soil solution of an acid forest soil Water extracts were prepared from soil samples which were collected from a soil profile showing very little variation in the texture down to a depth of 120 cm and thus only little translocation of clay in the soil profile. The aim of the study was to describe the distribution between soil and soil solution of several metals like Cu, Pb, Cd, Zn, Al and Mn as a function of humic substances, electrolyte concentration and pH. From the experimental results the following hypothesis on the reaction mechanisms involving metals and humus derived substances has been deduced. The metals Cu, Fe, Al and Pb are mobilized through complexation by soluble humus substances in addition to the usual pH dependent desorption and dissolution of hydroxides. This mobilization determines the solution concentration of Cu and Fe at pH > 3.7 and Al and Pb at pH > 4.2. Al, Fe and Pb are complexed selectively by high molecular weight humus derived substances which undergo adsorption on soil mineral surfaces. Cu interacts with low molecular weight humus derived substances which are not easily adsorbed by the mineral surfaces. Zn, Cd and Mn primarily undergo sorption and are thus controlled by pH and electrolyte concentration of solutions because their complexation with humus derived substances seems to be weak or nonexistant. It is further postulated that the humus derived substances mobilize Al³⁺ and Fe³⁺ ions. By this, other metals like Cd, Zn, Mn, Ca and Mg can occupy the free exchange sites.     

鹅细小病毒VP1与VP3非重叠序列的克隆与原核表达

根据鹅细小病毒GPVH1株基因组核苷酸序列设计了一对引物，对其结构蛋白VP1与VP3非重叠核苷酸序列进行PCR扩增，并克隆到表达性载体pGEX-6p-1，经酶切鉴定筛选出阳性克隆，并转化到大肠杆菌BL21(ED3)plysS进行原核表达，并用SDS—PAGE鉴定，结果表明融合蛋白在表达性细菌BL21中重获得了高效表达。