TaqMan PCR assay for detection and quantification of <Emphasis Type="Italic">Stagonosporopsis tanaceti</Emphasis> in pyrethrum seed and seedlings

Pyrethrum seed has an important role in the transmission of Stagonosporopsis tanaceti, the cause of ray blight disease of pyrethrum. A TaqMan probe based polymerase chain reaction (PCR) assay was developed to quantify the level of S. tanaceti inocula in pyrethrum seed and seedlings. Primer pair (St_qF3, St_qR2) was designed based on the intergenic spacer (IGS) region of S. tanaceti, which produced a 125 bp amplicon specific to S. tanaceti. TaqMan PCR assay using St_qF3, St_qR2 and a probe St_qP was highly specific against the genomic DNA of S. tanaceti, but did not amplify DNA of 14 related Stagonosporopsis species or other foliar pathogens of pyrethrum. The sensitivity limit of this assay was measured using the cycle threshold (C_t) value which ranged from 17.59 for 10 nanograms (ng) to 36.34 for 100 femtograms (fg) genomic DNA of S. tanaceti. There was a significant negative correlation (r = ?0.999, P < 0.001) between the C_t value and the percent of S. tanaceti infected seed. In addition, this TaqMan PCR assay detected latent infection within seedlings. This assay could be applied to test commercial seed and seedlings before deciding on the appropriate management practices.     

Ecoviability for ecosystem‐based fisheries management

Reconciling food security, economic development and biodiversity conservation is a key challenge, especially in the face of the demographic transition characterizing many countries in the world. Fisheries and marine ecosystems constitute a difficult application of this bio‐economic challenge. Many experts and scientists advocate an ecosystem approach to manage marine socio‐ecosystems for their sustainability and resilience. However, the ways by which to operationalize ecosystem‐based fisheries management (EBFM) remain poorly specified. We propose a specific methodological framework—viability modelling—to do so. We show how viability modelling can be applied using four contrasted case‐studies: two small‐scale fisheries in South America and Pacific and two larger‐scale fisheries in Europe and Australia. The four fisheries are analysed using the same modelling framework, structured around a set of common methods, indicators and scenarios. The calibrated models are dynamic, multispecies and multifleet and account for various sources of uncertainty. A multicriteria evaluation is used to assess the scenarios’ outcomes over a long time horizon with different constraints based on ecological, social and economic reference points. Results show to what extent the bio‐economic and ecosystem risks associated with the adoption of status quo strategies are relatively high and challenge the implementation of EBFM. In contrast, strategies called ecoviability or co‐viability strategies, that aim at satisfying the viability constraints, reduce significantly these ecological and economic risks and promote EBFM. The gains associated with those ecoviability strategies, however, decrease with the intensity of regulations imposed on these fisheries.     

Estimating symbiotic N2 fixation in Robinia pseudoacacia

Estimating symbiotic di‐nitrogen (N₂) fixation is challenging, especially when working with woody N₂ fixers in field trials. Fortunately, isotope methods based on ¹⁵N natural abundance or on ¹⁵N artificial enrichment (dilution method) make it possible to estimate the proportion of nitrogen derived from the atmosphere (Ndfa) in N₂‐fixing species. These methods have been extensively used in the field for herbaceous species, much less for tree species such as alder and acacia, and rarely for black locust (Robinia pseudoacacia). The objectives of this study were to characterize the fixation potential of black locust in a plantation by using the two ¹⁵N isotope methods in the field, and to document values of isotope fractionation occurring during N₂ fixation (the B value). B values were estimated both by growing trees on an N‐free medium in controlled conditions (B_lab) and by making Ndfa calculated with the natural abundance method converge with Ndfa calculated with the ¹⁵N dilution method in the field (B_field). The two methods gave consistent estimates of the B value. B values ranging between –1.4 and –3.2‰ were found, varying with the age of the plant material. Up to 76% of the N in the black locust trees came from the atmosphere, representing more than 45 kg N ha^?1 over five years and confirming that black locust may be well adapted to N‐poor soils.     

Sublethal effects of nine insecticides on Drosophila suzukii and its major pupal parasitoid Trichopria drosophilae

Background

Although the pupal parasitoid Trichopria drosophilae is used in conservative and augmentative biocontrol of Drosophila suzukii infestations, current pest management strategies mostly rely on multiple insecticide applications. In this context, the aim of the study was to investigate the baseline toxicity of nine insecticides on D. suzukii larvae and their multiple sublethal effects (LC₁₀) on immature stages of the pest feeding on contaminated diet and T. drosophilae developing within the intoxicated host.

Results

Chlorpyriphos and azadirachtin showed the lowest and the highest LC₁₀, the values of which were 9.78 × 10¹³ and 1.46 × 10³ times lower than their recommended label field rate, respectively. Among tested insecticides, imidacloprid, malathion and dimethoate were the only treatments that did not affect the juvenile development time of D. suzukii, while spinosad and the organophosphates chlorpyriphos and dimethoate did not influence fly pupal size. No sublethal effects were recorded on T. drosophilae degree of infestation (DI) and juvenile development time. On the contrary, cyazypyr and dimethoate negatively affected the success of parasitism (SP) and the number of progeny of the pupal parasitoid, in association with malathion for the first parameter and spinosad for the fertility. Compared to the untreated control, more female progeny emerged following azadirachtin exposure, while dimethoate caused the opposite effect. Imidacloprid, lambda-cyhalothrin and spinetoram decreased hind tibia length of emerged parasitoids.

Conclusion

This study provides new insights on the (eco)toxicological profile of nine insecticides and new information needed to support the deployment of T. drosophilae in the field within the sustainable management techniques against D. suzukii. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Population pharmacokinetics of methadone hydrochloride after a single intramuscular administration in adult Japanese sika deer (Cervus nippon nippon)

ObjectiveTo assess the population pharmacokinetics of methadone in deer.Study designProspective non-randomized experimental trial.AnimalsTwelve healthy adult sika deer (nine males and three females).MethodsDeer received intramuscular administration of racemic methadone hydrochloride at 0.5 mg kg⁻¹ or 1 mg kg⁻¹. Plasma methadone and its metabolite 2-Ethylidene-1,5-Dimethyl-3,3-Diphenyl-Pyrolidine (EDDP) concentrations were determined by validated liquid chromatography coupled to tandem mass spectrometry methods, at times 0, 30 minutes, 1, 2, 3, 4, 5, 6, 8, 12 and 24 hours. Population pharmacokinetics analysis was undertaken using a non-linear mixed effects modelling (NONMEM).ResultsA two-compartment linear disposition model best described observed time-concentration profiles of methadone and EDDP. Population parameter estimates of methadone were elimination clearance (17.3 L hour⁻¹), metabolic clearance (34.6 L hour⁻¹), volume of distribution of compartment 1 (216.0 L) and volume of distribution of compartment 2 (384.0 L). Population parameter estimates of EDDP were elimination clearance (121.0 L hour⁻¹), volume of distribution of compartment 3 (1.08 L) and volume of distribution of compartment 4 (499.5 L). The total clearance and total volume of distribution of methadone and EDDP were 51.9 L hour⁻¹, 121.0 L hour ⁻¹, 600.0 L and 500.6 L, respectively. The methadone terminal elimination half-life was 8.19 hours. No adverse effects were observed after methadone administration.Conclusions and Clinical relevanceFollowing intramuscular injection, methadone was characterized by a large total volume of distribution, high systemic clearance and intermediate terminal half-life in sika deer.     

Hepatitis E virus chronic infection of swine co-infected with Porcine Reproductive and Respiratory Syndrome Virus

In developed countries, most of hepatitis E human cases are of zoonotic origin. Swine is a major hepatitis E virus (HEV) reservoir and foodborne transmissions after pork product consumption have been described. The risk for HEV-containing pig livers at slaughter time is related to the age at infection and to the virus shedding duration. Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is a virus that impairs the immune response; it is highly prevalent in pig production areas and suspected to influence HEV infection dynamics. The impact of PRRSV on the features of HEV infections was studied through an experimental HEV/PRRSV co-infection of specific-pathogen-free (SPF) pigs. The follow-up of the co-infected animals showed that HEV shedding was delayed by a factor of 1.9 in co-infected pigs compared to HEV-only infected pigs and specific immune response was delayed by a factor of 1.6. HEV shedding was significantly increased with co-infection and dramatically extended (48.6 versus 9.7 days for HEV only). The long-term HEV shedding was significantly correlated with the delayed humoral response in co-infected pigs. Direct transmission rate was estimated to be 4.7 times higher in case of co-infection than in HEV only infected pigs (0.70 and 0.15 per day respectively). HEV infection susceptibility was increased by a factor of 3.3, showing the major impact of PRRSV infection on HEV dynamics. Finally, HEV/PRRSV co-infection – frequently observed in pig herds – may lead to chronic HEV infection which may dramatically increase the risk of pig livers containing HEV at slaughter time.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-015-0207-y) contains supplementary material, which is available to authorized users.     

Intra‐household constraints on educational attainment in rural households in Papua New Guinea

This paper examines intra‐household and socio‐cultural factors leading to differential outcomes in educational attainment by gender and birth order amongst smallholder oil palm households in Papua New Guinea. Not all children share equitably in the household resources allocated to education: females have lower average education levels than males, and high birth order children have higher education attainment than lower birth order siblings, indicating preferential parental investment in sons and early born children. The findings demonstrate that despite households having regular access to relatively high incomes from oil palm and residing in close proximity to schools, primary school net enrolment rates remain significantly lower than those for East Asia and the Pacific region, and the millennium development goal of universal primary education has not been met. This finding is likely to be the result of a combination of intra‐household factors including gender power imbalances, low parental education levels, the agency of youth in educational decision‐making and the weakening attraction of education as a means of improving income‐earning potential.     

Heparanase and Syndecan-4 Are Involved in Low Molecular Weight Fucoidan-Induced Angiogenesis

Induction of angiogenesis is a potential treatment for chronic ischemia. Low molecular weight fucoidan (LMWF), the sulfated polysaccharide from brown seaweeds, has been shown to promote revascularization in a rat limb ischemia, increasing angiogenesis in vivo. We investigated the potential role of two heparan sulfate (HS) metabolism enzymes, exostosin-2 (EXT2) and heparanase (HPSE), and of two HS-membrane proteoglycans, syndecan-1 and -4 (SDC-1 and SDC-4), in LMWF induced angiogenesis. Our results showed that LMWF increases human vascular endothelial cell (HUVEC) migration and angiogenesis in vitro. We report that the expression and activity of the HS-degrading HPSE was increased after LMWF treatment. The phenotypic tests of LMWF-treated and EXT2- or HPSE-siRNA-transfected cells indicated that EXT2 or HPSE expression significantly affect the proangiogenic potential of LMWF. In addition, LMWF increased SDC-1, but decreased SDC-4 expressions. The effect of LMWF depends on SDC-4 expression. Silencing EXT2 or HPSE leads to an increased expression of SDC-4, providing the evidence that EXT2 and HPSE regulate the SDC-4 expression. Altogether, these data indicate that EXT2, HPSE, and SDC-4 are involved in the proangiogenic effects of LMWF, suggesting that the HS metabolism changes linked to LMWF-induced angiogenesis offer the opportunity for new therapeutic strategies of ischemic diseases.     

Genetic mapping,marker assisted selection and allelic relationships for the Pu6 gene conferring rust resistance in sunflower

Rust resistance in the sunflower line P386 is controlled by Pu₆, a gene which was reported to segregate independently from other rust resistant genes, such as R₄. The objectives of this work were to map Pu₆, to provide and validate molecular tools for its identification, and to determine the linkage relationship of Pu₆ and R₄. Genetic mapping of Pu₆ with six markers covered 24.8 cM of genetic distance on the lower end of linkage Group 13 of the sunflower consensus map. The marker most closely linked to Pu₆ was ORS316 at 2.5 cM in the distal position. ORS316 presented five alleles when was assayed with a representative set of resistant and susceptible lines. Allelism test between Pu₆ and R₄ indicated that both genes are linked at a genetic distance of 6.25 cM. This is the first confirmation based on an allelism test that at least two members of the R_adv/R₄/R₁₁/ R_13a/R_13b/Pu₆ cluster of genes are at different loci. A fine elucidation of the architecture of this complex locus will allow designing and constructing completely new genomic regions combining genes from different resistant sources and the elimination of the linkage drag around each resistant gene.