Resistance to infection by stealth: <Emphasis Type="Italic">Brassica napus</Emphasis> (winter oilseed rape) and <Emphasis Type="Italic">Pyrenopeziza brassicae</Emphasis> (light leaf spot)

Light leaf spot (Pyrenopeziza brassicae) is an important disease on winter oilseed rape crops (Brassica napus) in northern Europe. In regions where economically damaging epidemics occur, resistance to P. brassicae in commercial cultivars is generally insufficient to control the disease without the use of fungicides. Two major genes for resistance have been identified in seedling experiments, which may operate by decreasing colonisation of B. napus leaf tissues and P. brassicae sporulation. Much of the resistance present in current commercial cultivars is thought to be minor gene-mediated and, in crops, disease escape and tolerance also operate. The subtle strategy of the pathogen means that early colonisation of host tissues is asymptomatic, so a range of techniques and molecular tools is required to investigate mechanisms of resistance. Whilst resistance of new cultivars needs to be assessed in field experiments where they are exposed to populations of P. brassicae under natural conditions, such experiments provide little insight into components of resistance. Genetic components are best assessed in controlled environment experiments with single spore (genetically fixed) P. brassicae isolates. Data for cultivars used in the UK Recommended List trials over several seasons demonstrate how the efficacy of cultivar resistance can be reduced when they are deployed on a widespread scale. There is a need to improve understanding of the components of resistance to P. brassicae to guide the development of breeding and deployment strategies for sustainable management of resistance to P. brassicae in Europe.     

Ophiotaenia bonneti sp. n. (Eucestoda: Proteocephalidea), a parasite of Rana vaillanti (Anura: Ranidae) in Costa Rica

Ophiotaenia bonneti sp. n. is described from the intestine of the frog Rana vaillanti Brocchi, 1877 (Anura: Ranidae) from San Gerardo, Guanacaste, Costa Rica. The new species is characterized by the testes 100-177 in number, the genital pores situated anteriorly, the osmoregulatory canals overlapping the testis field, the cirrus pouch length as 15-24% of proglottis width, and the uterus with 18-32 ramified diverticula on each side. It differs from the 23 known species of the genus Ophiotaenia La Rue, 1911, parasitic in amphibians, by one to several morphological characters. It differs from O. gracilis Jones, Cheng et Gillespie, 1958, the most morphologically similar species, in the sucker diameter in % of scolex diameter and in the morphology of the eggs - funnel-like depression and embryophore closely investing the oncosphere in O. gracilis. We generally observe a very low mean prevalence of the Proteocephalidea in Neotropical amphibians (about 0.41%-3%), but in the case of some host species, the prevalence can reach up to 25%. We conclude that these cestodes exhibit a strict host specificity of the oioxene type. Ophiotaenia junglensis Srivastava et Capoor, 1980 is considered a species inquirenda. Batrachotaenia hernandezi (Flores-Barroeta, 1955) becomes Ophiotaenia hernandezi (Flores-Barroeta, 1955) comb. n., B. tigrina (Woodland, 1925) becomes O. tigrina (Woodland, 1925) comb. n. and B. ceratophryos (Parodi et Widakowich, 1916) becomes O. ceratophryos (Parodi et Widakowich, 1916) comb. n.     

Transformation of Eutypa dieback and esca disease pathogen toxins by antagonistic fungal strains reveals a second detoxification pathway not present in Vitis vinifera

Eutypine, 4-hydroxybenzaldehyde, and 3-phenyllactic acid are some of the phytotoxins produced by the pathogens causing Eutypa dieback and esca disease, two trunk diseases of grapevine (Vitis vinifera). Known biocontrol agents such as Fusarium lateritium and Trichoderma sp. were screened for their ability to consume these toxins. Transformation time courses were performed, and an high-performance liquid chromatography-based method was developed to analyze toxin metabolism and to identify and quantify the converted products. The results show that the aldehyde function of eutypine was reduced to eutypinol, as by V. vinifera cv. Merlot, the cultivar tolerant to Eutypa dieback. We revealed a supplementary detoxification pathway, not known in Merlot, where the aldehyde function was oxidized to eutypinic acid. Moreover, some strains tested could further metabolize the transformation products. Every strain tested could transform 4-hydroxybenzaldehyde to the corresponding alcohol and acid, and these intermediates disappeared totally at the end of the time courses. When biological assays on cells of V. vinifera cv. Chasselas were carried out, the transformation products exhibited a lower toxicity than the toxins. The possibility of selecting new biocontrol agents against trunk diseases of grapevine based on microbial detoxification is discussed.     

Radial variations of vibrational properties of three tropical woods

The radial trends of vibrational properties, represented by the specific dynamic modulus (E′/ρ) and damping coefficient (tan δ), were investigated for three tropical rainforest hardwood species (Simarouba amara, Carapa procera, and Symphonia globulifera) using free-free flexural vibration tests. The microfibril angle (MFA) was estimated using X-ray diffraction. Consistent patterns of radial variations were observed for all studied properties. E′/ρ was found to decrease from pith to bark, which was strongly related to the increasing pith-bark trend of MFA. The variation of tan δ along the radius could be partly explained by MFA and partly by the gradient of extractives due to heartwood formation. The coupling effect of MFA and extractives could be separated through analysis of the log(tan δ) versus log(E′/ρ) diagram. For the species studied, the extractive content putatively associated with heartwood formation generally tends to decrease the wood damping coefficient. However, this weakening effect of extractives was not observed for the inner part of the heartwood, suggesting that the mechanical action of extractives was reduced during their chemical ageing.     

Evaluation of the anaesthetic effects of combinations of ketamine, medetomidine, romifidine and butorphanol in European badgers (Meles meles)

OBJECTIVE: To evaluate the effects of three anaesthetic combinations in adult European badgers (Meles meles). STUDY DESIGN: Prospective, randomized, blinded, experimental trial. ANIMALS: Sixteen captive adult badgers. METHODS: The badgers were each anaesthetized by intramuscular injection using the three techniques assigned in random order: romifidine 0.18 mg kg(-1), ketamine 10 mg kg(-1) and butorphanol 0.1 mg kg(-1) (RKB); medetomidine 0.1 mg kg(-1), ketamine 9 mg kg(-1) and butorphanol 0.1 mg kg(-1) (MKB); and medetomidine 0.1 mg kg(-1) and ketamine 10 mg kg(-1) (MK). Initial drug doses were calculated based on a body mass of 10 kg. Additional anaesthetic requirements, time to drug effect, duration of action and recovery from anaesthesia were recorded. Heart rate and rhythm, respiratory rate and rhythm, rectal and subcutaneous microchip temperature and oxygen saturation were recorded every 5 minutes. Depth of anaesthesia was assessed using: muscle tone; palpebral and pedal reflexes; and tongue relaxation at these time points. Blood samples and a tracheal aspirate were obtained under anaesthesia. Atipamezole was administered if the badger had not recovered within 60 minutes Parametric data were analysed using anova for repeated measures, and nonparametric data using Friedman's, and Cochran's Q tests: p < 0.05 was considered significant. RESULTS: All combinations produced good or excellent muscle relaxation throughout the anaesthetic period. RKB had the shortest duration of anaesthesia (16.8 minutes compared with MKB 25.9 minutes and MK 25.5 minutes) and antagonism was not required. RKB depressed respiratory rate less than MK and MKB. There was no significant difference between techniques for heart rate and rhythm. CONCLUSIONS AND CLINICAL RELEVANCE: All combinations provided anaesthetic conditions suitable for sampling and identification procedures in adult badgers. The RKB protocol provided a significantly shorter period of anaesthesia when compared with the combinations containing medetomidine.     

Effect of habitat spatiotemporal structure on collembolan diversity

Landscape fragmentation is a major threat to biodiversity. It results in the transformation of continuous (hence large) habitat patches into isolated (hence smaller) patches, embedded in a matrix of another habitat type. Many populations are harmed by fragmentation because remnant patches do not fulfil their ecological and demographic requirements. In turn, this leads to a loss of biodiversity, especially if species have poor dispersal abilities. Moreover, landscape fragmentation is a dynamic process in which patches can be converted from one type of habitat to another. A recently created habitat might suffer from a reduced biodiversity because of the absence of adapted species that need a certain amount of time to colonize the new patch (e.g. direct meta-population effect). Thus landscape dynamics lead to complex habitat spatiotemporal structured, in which each patch is more or less continuous in space and time. In this study, we define habitat spatial structure as the degree to which a habitat is isolated from another habitat of the same kind and temporal structure as the time since the habitat is in place. Patches can also display reduced biodiversity because their spatial or temporal structures are correlated with habitat quality (e.g. indirect effects). We discriminated direct meta-community effects from indirect (habitat quality) effects of the spatiotemporal structure of habitats on biodiversity using Collembola as a model. We tested the relative importance of spatial and temporal structure of habitats for collembolan diversity, taking soil properties into account. In an agroforested landscape, we set up a sampling design comprised of two types of habitats (agriculture versus forest), a gradient of habitat isolation (three isolation classes) and two contrasting ages of habitats. Our results showed that habitat temporal structure is a key factor shaping collembolan diversity. A reduced diversity was detected in recent habitats, especially in forests. Interactions between temporal continuity and habitat quality were also detected by taking into account soil properties: diversity increased with soil carbon content, especially in old forests. Negative effects of habitat age on diversity were stronger in isolated patches. We conclude that habitat temporal structure is a key factor shaping collembolan diversity, while direction and amplitude of its effect depend on land use type and spatial isolation.     

Annual dynamics of Zymoseptoria tritici populations in wheat cultivar mixtures: A compromise between the efficacy and durability of a recently broken-down resistance gene?

Cultivar mixtures slow polycyclic epidemics but may also affect the evolution of pathogen populations by diversifying the selection pressures exerted by their plant hosts at field scale. We compared the dynamics of natural populations of the fungal pathogen Zymoseptoria tritici in pure stands and in three binary mixtures of wheat cultivars (one susceptible cultivar and one cultivar carrying the recently broken-down Stb16q gene) over two annual field epidemics. We combined analyses of population “size” based on disease severity, and of population “composition” based on changes in the frequency of virulence against Stb16q in seedling assays with more than 3000 strains. Disease reductions were observed in mixtures late in the epidemic, at the whole-canopy scale and on both cultivars, suggesting the existence of a reciprocal protective effect. The three cultivar proportions in the mixtures (0.25, 0.5, and 0.75) modulated the decrease in (a) the size of the pathogen population relative to the two pure stands, (b) the size of the virulent subpopulation, and (c) the frequency of virulence relative to the pure stand of the cultivar carrying Stb16q. Our findings suggest that optimal proportions may differ slightly between the three indicators considered. We argue potential trade-offs that should be taken into account when deploying a resistance gene in cultivar mixtures: between the dual objectives “efficacy” and “durability,” and between the “size” and “frequency” of the virulent subpopulation. Based on current knowledge, it remains unclear whether virulent subpopulation size or frequency has the largest influence on interepidemic virulence transmission.     

Immunogenic and protective properties of GP5 and M structural proteins of porcine reproductive and respiratory syndrome virus expressed from replicating but nondisseminating adenovectors

Porcine reproductive and respiratory syndrome virus (PRRSV) is responsible for significant economic losses in the porcine industry. Currently available commercial vaccines do not allow optimal and safe protection. In this study, replicating but nondisseminating adenovectors (rAdV) were used for the first time in pigs for vaccinal purposes. They were expressing the PRRSV matrix M protein in fusion with either the envelope GP5 wild-type protein (M-GP5) which carries the major neutralizing antibody (NAb)-inducing epitope or a mutant form of GP5 (M-GP5m) developed to theoretically increase the NAb immune response. Three groups of fourteen piglets were immunized both intramuscularly and intranasally at 3-week intervals with rAdV expressing the green fluorescent protein (GFP, used as a negative control), M-GP5 or M-GP5m. Two additional groups of pigs were primed with M-GP5m-expressing rAdV followed by a boost with bacterially-expressed recombinant wild-type GP5 or were immunized twice with a PRRSV inactivated commercial vaccine. The results show that the rAdV expressing the fusion proteins of interest induced systemic and mucosal PRRSV GP5-specific antibody response as determined in an ELISA. Moreover the prime with M-GP5m-expressing rAdV and boost with recombinant GP5 showed the highest antibody response against GP5. Following PRRSV experimental challenge, pigs immunized twice with rAdV expressing either M-GP5 or M-GP5m developed partial protection as shown by a decrease in viremia overtime. The lowest viremia levels and/or percentages of macroscopic lung lesions were obtained in pigs immunized twice with either the rAdV expressing M-GP5m or the PRRSV inactivated commercial vaccine.     

Effect of timing and duration of salt treatment during growth of a fragrant rice variety on yield and 2-acetyl-1-pyrroline, proline, and GABA Levels

In greenhouse experiments, Aychade, a fragrant rice variety, was grown under one level of salt solution (EC of 3800 ± 400 μS·cm(-1)) sufficient to induce salt stress in rice. Timing and duration of salt solution application varied according to the growth stages. 2-Acetyl-1-pyrroline (2AP), a characteristic flavor compound of fragrant rice as well as biogenetically related compounds, proline, and γ-aminobutyric acid (GABA) were quantified. Salt treatments induced 2AP synthesis in the leaves, but the increase was often higher in the vegetative phase. This increase was correlated with proline level but not with that of GABA. Interestingly the grains from all the salt treated plants contained significantly higher levels of 2AP (733-998 μg·kg(-1)) than those from the control (592 μg·kg(-1)). The highest 2AP synthesis occurred when the plants were subjected to salt treatment during whole vegetative or reproductive phases. However in the latter case crop yield decreased significantly.