Differential Activity of Carboxylic Acid Amide Fungicides Against Various Developmental Stages of Phytophthora infestans

ABSTRACT Three carboxylic acid amide (CAA) fungicides, mandipropamid (MPD), dimethomorph (DMM) and iprovalicarb (IPRO) were examined for their effects on various asexual developmental stages of Phytophthora infestans in vitro and in planta. Germination of cystospores and direct germination of sporangia were inhibited with nanomole concentrations of MPD (0.005 mug/ml) and micromole concentrations of DMM (0.05 mug/ml) or IPRO (0.5 mug/ml). A temporary exposure of 1 h to CAAs was not detrimental to germination and infectivity of sporangia or cystospores. CAAs applied to cystospores at 1 h after the onset of germination did not prevent the emergence of germ tubes, but inhibited their further growth and deformed their shape. None of the fungicides affected discharge of zoospores from sporangia or the encystment (cell wall formation/assembly) of the zoospores. Mycelium growth in solid or liquid media was inhibited with micromole concentrations. CAAs mixed with sporangia and drop inoculated onto detached leaves strongly suppressed infection. Curative application at 1 day postinoculation (dpi) required higher concentrations of CAAs than preventive application to inhibit infection and lost its effectiveness at 2 dpi. When sprayed on established late blight lesions 4 days after inoculation, CAAs reduced sporangial production in a dose-dependent manner. Trans-laminar protection of potato or tomato leaves, although achieved with higher doses, was more effective with MPD than with DMM or IPRO. Shade house studies demonstrated superior control of late blight epidemics by MPD compared with the other molecules. The data suggest that germ tube formation by cystospores or sporangia is the most sensitive stage in the life cycle of P. infestans to CAAs. Of the three CAAs, MPD had the highest intrinsic activity against spore germination. This property, together with its better trans-laminar activity, makes MPD more effective than DMM or IPRO in controlling epidemics caused by P. infestans.     

After nasopharyngeal infection,foot-and-mouth disease virus serotype A RNA is shed in bovine milk without associated mastitis

Foot-and-mouth disease (FMD) is a highly contagious aphthoviral infection of cloven-hoofed animals, inducing vesiculopustular stomatitis, pododermatitis, and thelitis. Vesicular fluid represents a major pathway of virus excretion, but bovine milk is another important source of virus shedding. We describe here the time course of FMD virus (FMDV) excretion in the milk and characterize associated lesions in the mammary gland. Three dairy cows were infected by nasopharyngeal instillation of FMDV and monitored over 12 d. Autopsy was performed at the end of the study, and specimens were collected for histopathology, IHC, and RT-qPCR. All 3 cows developed fever, drooling, vesiculopustular stomatitis, interdigital dermatitis, and thelitis. FMDV RNA was detectable in whole milk until the end of the trial, but only transiently in saliva, nasal secretions, and blood serum. Although histology confirmed vesiculopustular lesions in the oral and epidermal specimens, the mammary glands did not have unequivocal evidence of FMDV-induced inflammation. FMDV antigen was detectable in skin and oral mucosa, but not in the mammary gland, and FMDV RNA was detectable in 9 of 29 samples of squamous epithelia but only in 1 of 12 samples of mammary gland.     

Impact of temperature and nutrition on the toxicity of the insecticide λ-cyhalothrin in full-lifecycle tests with the target mosquito species Aedes albopictus and Culex pipiens

The global spread of the Asian tiger mosquito Aedes albopictus, an urban pest as well as a vector for arboviruses, is a threat for public health. As control measures include the use of insecticides such as the pyrethroid λ-cyhalothrin, it is crucial to assess their efficiency and their potential impact on the biodiversity especially under climate change conditions. To evaluate the environmental risk, biotests are well established for non-target organisms but not yet for mosquitoes. We therefore developed a full-lifecycle biotest for mosquitoes kept under quarantine conditions based on the OECD guideline 219. Therewith we tested the effect of temperature and nutrition on the ecotoxicological response to λ-cyhalothrin on the mosquitoes Ae. albopictus and Culex pipiens by assessing sublethal and life history parameters. The efficiency of λ-cyhalothrin decreased in both mosquito species with increasing temperature and changed with feeding protocol. At effective concentrations for potential mosquito control in surface waters, λ-cyhalothrin poses a high risk for indigenous aquatic key role species inhabiting the same microhabitats. Those aspects should to be taken into account in vector control strategies.     

Habitat structure and prey aggregation determine the functional response in a soil predator-prey interaction

Functional responses describe the per capita consumption rates of predators depending on prey density, which quantifies the energy transfer between trophic levels. We studied a typical interaction of the litter-soil systems between hunting spiders (Pardosa lugubris; Araneae: Lycosidae) and springtails (Heteromurus nitidus; Collembola: Entomobryidae) at varying habitat structure, i.e. with moss vs. without moss. We found a hyperbolic increase in consumption (functional response type II) in the treatment without habitat structure that was converted into a roller-coaster (or dome-shaped in a broad sense) functional response in treatments with habitat structure. Additional experiments suggest that the reduced per capita consumption rates at high prey densities may be explained by aggregative defence behaviour of the springtails. Experimentally, this behaviour was induced by the presence of habitat structure. We analyzed the net-energy gain of this predator-prey interaction by comparing the predator’s metabolic energy loss to its energy gain by consumption. In treatments with habitat structure, the net-energy gain of the predator was limited at intermediate prey densities where prey aggregation reduced the consumption rates. Our results stress the importance of habitat structure and prey behaviour in shaping the functional response in a typical soil-litter predator-prey interaction.     

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.     

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.