Measurement of systemic and local respiratory cell-mediated immunity after influenza infection in chickens

The detection of ChIFN production after ex vivo antigenic-stimulation of T-lymphocytes has been evaluated for the first time, as a tool to assess cell-mediated immunity (CMI) after avian influenza (AI) infection in 10-day-old SPF chickens. Preliminarily, recall antigens have been produced either by concentrating and inactivating the whole virus or by dissociating the viral proteins. Biologically and structurally intact forms of the viral proteins were isolated by non-ionic detergents while heating, chemical agents and ionic detergent used for virus inactivation altered the antigenic viral components. The n-octyl-B-D-gluco-pyranoside treatment at low temperature was very efficient to produce AI antigenic proteins used for evaluation of ChIFN production after ex vivo antigenic-stimulation of splenic and peripheral lymphocytes. In addition, protocols to isolate lymphocytes from the respiratory tract - the trachea and the lung - have been adapted for local CMI evaluation after similar ex vivo recall assay. Specific AI CMI in the spleen, the blood and the lung was detected for 5 weeks after low pathogenic AI (LPAI) infection in chickens, while further development is needed for tracheal CMI measurement.     

Uptake and utilisation of phosphonate ions by Phytophthora citrophthora and Nectria haematococca in relation to their selective toxicity

Phosphonate ions, which are degradation products of aluminium tris-O-ethyl phosphonate (fosetyl-aluminium) in plant tissues, had more effect in vitro against Phytophthora citrophthora than against Nectria haematococca. Both fungi were able to take up phosphonate ions. This uptake was realised against a concentration gradient and was affected by metabolic inhibitors. When examined over a 0.5 to 6.0 mm concentration range, phosphonate was carried by a single transport process obeying Michaelis-Menten kinetics, with a low affinity (high K′_m). Further analysis by means of Hofstee plots indicated a somewhat faster phosphonate uptake by N. haematococca than by P. citrophthora, so it appears clearly that uptake was not involved in the selective toxicity of this compound. The continuous uptake of phosphonate by N. haematococca is correlated with an important utilisation of this ion by the fungal cells. This fact could be explained by a detoxification process corresponding to the oxidation of phosphonate to phosphate which could account for the low sensitivity of this fungus. Phosphonate uptake by P. citrophthora reached a maximum level after 20 min and then decreased to a constant level over the following period. During the incubation period phosphonate was incorporated directly without oxidation into phosphate. This incorporation could lead to a disturbance of the cell metabolism.     

Diversity of root-knot nematodes in Moroccan olive nurseries and orchards: does <Emphasis Type="Italic">Meloidogyne javanica</Emphasis> disperse according to invasion processes?

Background

Root-knot nematodes (RKN) are major pest of olive tree (Olea europaea ssp. europaea), especially in nurseries and high-density orchards. Soil samples were collected from main olive growing areas of Morocco, to characterize Meloidogyne species and to discuss the contribution of biotic and abiotic factors in their spatial distribution.

Results

RKN were found in 159 soil samples out of 305 from nurseries (52.1% occurrence) and in 11 out of 49 soil samples from orchards (23.2% occurrence). Biochemical and molecular characterisation (PAGE esterase and SCAR) revealed the dominance of M. javanica both in nurseries and orchards with minor presence of M. incognita only in nurseries, and M. arenaria in only one nursery. RKN were distributed on aggregated basis. Frequent presence of M. javanica in orchards might have come from nurseries. In contrast, the detection of M. incognita in nurseries alone suggests that this species could not reproduce in orchards because of either the competition with other plant-parasitic nematodes or unfit local habitats. The impact of environmental variables (climate, habitat origin and physicochemical characteristics of the substrates) on the distribution of Meloidogyne species is also discussed.

Conclusion

Olive nurseries in Morocco are not able to guarantee the safety of rooted plants. As a result, olive production systems are exposed to strong RKN invasion risks. Consequently, the use of healthy substrates in nurseries may prevent plant-parasitic nematode induction in orchards.

     

Challenges of thermal nociceptive threshold testing in the donkey

ObjectiveTo evaluate a thermal nociceptive threshold (TNT) testing device in the donkey, and the influence of potential confounding factors on TNTs.AnimalsTwo groups (Group 1 and Group 2) of eight castrated male donkeys aged 4–9 years, weighing 105–170 kg.MethodsTNTs were measured by heating a thermal probe on skin until an end-point behaviour (threshold temperature) or a cut-out temperature (51 °C) was reached. The withers and the dorsal aspect of the distal limb were used as sites for TNT testing. The effects on TNT of different confounding factors: the limb tested; rate of heating; and ambient temperature were evaluated. Data were analyzed using general linear models, and Mann-Whitney tests, p < 0.05 was considered significant.ResultsEnd-point behaviours (skin twitch or donkey looking at test device) when the thermal probe heated the withers were observed in approximately half of tests. TNT was (mean ± SD) 46.8 ± 2.85 °C. Subsequently the limb was evaluated as the test site in Group 1 followed by Group 2 donkeys; end-point behaviour being a foot-lift. In Group 1, 72% of tests ended in an end-point behaviour but the response rate was lower in Group 2 (20%), although TNTs were similar [(47.6 ± 3.3) and (47.3 ± 3.0) °C respectively] for responding animals. Rate of heating, ambient temperature and laterality (right or left) did not affect thresholds, but mean TNT was significantly higher in the forelimb (48.5 ± 2.8 °C) than the hind limb (47.4 ± 2.8 °C) (p = 0.012).ConclusionsWhen a thermal probe cut-out temperature of 51 °C was used in TNT testing in the donkey a high proportion of tests did not produce an identifiable end point behaviour. Higher cut-out temperatures damaged the skin. Under these conditions, thermal nociceptive threshold testing appears not be an appropriate analgesiometry technique in the donkey.Clinical relevanceTNT testing under these conditions is not suitable form of analgesiometry for donkeys.     

The influence of various confounding factors on mechanical nociceptive thresholds in the donkey

ObjectiveTo evaluate a mechanical nociceptive threshold (MNT) testing device in the donkey, and to investigate the influence of potential confounders on MNTs generated.Study designProspective, randomised.AnimalsSixteen castrated male donkeys aged 4–9 years, weighing 105–170 kg.MethodsMechanical nociceptive thresholds were measured using an actuator with three pins placed on the dorsal aspect of the distal limb, connected to a force meter. The pins (surface area 15 mm²) were extruded onto the limb by pressurising an air-filled syringe, until the MNT force (when foot-lift was observed) or 25 N (cut-off force) was reached. Effect on MNT of presence of a companion donkey, the limb tested, rate of application of force, testing location, level of distraction, ambient temperature and hair cover at the test site was evaluated. Long and short-term repeatability of MNT was assessed. Data were analysed using general linear models and Mann–Whitney U tests, p < 0.05 was considered significant.ResultsIncreasing the rate of force application significantly increased the mean ± SD MNT from 9.2 ± 2.0 N when applied at 0.4 N sec^?1 to 10.6 ± 2.1 N when applied at 1.2 N sec^?1 (p = 0.001). No other factors significantly influenced MNT. Mean MNT remained stable over a 3 week period, however MNTs were significantly (p = 0.006) higher (12.8 ± 3.0 N cf 10.3 ± 1.9 N) after a 12 month interval.Conclusions and clinical relevanceWhen designing studies measuring MNT in donkeys, rate of application of force must be standardised. Donkeys’ MNTs have good short-term stability suggesting this technique is appropriate for short-term analgesiometry studies; however variability of MNTs over the long-term is greater.     

Biodiversity,ecology, fisheries,and use and trade of Tetraodontiformes fishes reveal their socio‐ecological significance along the tropical Brazilian continental shelf

1. Tetraodontiformes fishes play a critical role in benthic and demersal communities and are facing threats due to anthropogenic impacts and climate change. However, they are poorly studied worldwide. To improve knowledge on the socio‐ecological significance and conservation of Tetraodontiformes a review of literature addressing the diversity, ecology, use and trade, conservation, and main threats of Tetraodontiformes combined with a comprehensive in situ dataset from two broad‐range multidisciplinary oceanographic surveys performed along the Tropical Brazilian Continental Shelf was undertaken.
2. Twenty‐nine species were identified, being primarily found on coral reefs and algal ecosystems. At these habitats, tetraodontids present highly diversified trophic categories and might play an important role by balancing the marine food web
3. Coral reef ecosystems, especially those near to the shelf break, seem to be the most important areas of Tetraodontiformes fishes, concentrating the highest values of species richness, relative abundance and the uncommon and Near Threatened species.
4. Ninety per cent of species are commonly caught as bycatch, being also used in the ornamental trade (69%) and as food (52%), serving as an important source of income for artisanal local fisheries.
5. Tetraodontiformes are threatened by unregulated fisheries, overexploitation, bycatch, and habitat loss due to coral reef degradation and the potential effects of climate change. These factors are more broadly impacting global biodiversity, food security, and other related ecosystem functions upon which humans and many other organisms rely.
6. We recommend the following steps that could improve the conservation of Tetraodontiformes along the tropical Brazilian Continental shelf and elsewhere: (i) data collection of the commercial, incidental, ornamental and recreational catches; (ii) improvement of the current legislation directed at the marine ornamental harvesting; (iii) increase efforts focused on the education and conservation awareness in coastal tourism and communities; and, most important, (iv) creation of marine reserves networks in priority areas of conservation, protecting either the species and key habitats for its survival.

     

Dealing with the variability in biofumigation efficacy through an epidemiological framework

Biofumigation, as originally defined, is the use, in agriculture, of the toxicity of Brassica crop residues to control soilborne plant pathogens. This toxicity is specifically attributed to the release of toxic isothiocyanates, through the hydrolysis of glucosinolates present in the crop residues. This technique is considered a possible alternative to the use of pesticides, but field studies have generated conflicting data concerning the efficacy of biofumigation at field scale, limiting the use of this technique. Analytical studies based on a systematic approach involving evaluation of the potential effects of isothiocyanates can be used to address this problem in a rigorous manner. However, many recent studies have indicated that the mechanisms underlying biofumigation are much more complex than a simple toxic effect of residues. In this review, we dissect and discuss the problems encountered when trying to understand the variability in biofumigation efficacy and propose an integrative epidemiological approach to overcome these problems. This approach involves separating the effects of the different parameters of the system, such as the effects of different management phases of the biofumigant crop (i.e. the period of biofumigant crop growth and the phase during which crop residues are pulverised and incorporated into the soil) on the epidemiological mechanisms driving the development of an epidemic (density of primary inoculum and dynamics of disease progression). Finally, we propose new avenues of research into biofumigation in which the use of epidemiological tools and methods may improve our understanding of the factors underlying variation in the efficacy of biofumigant crops.     

Effect of Geogenic Lead on Fungal and Collembolan Communities in Garden Topsoil

Geogenic lead (Pb) is considered to be less bioavailable than anthropogenic Pb and exerts less effect on the soil fauna. However, Pb contamination in vegetables has been reported in the case of geogenic anomalies, even at moderate concentrations (around 170 mg kg^–1). In this study, we investigated collembolan communities using both taxonomic- and trait-based approaches and observed fungal communities to assess the effects of a moderate geogenic Pb anomaly on collembolans and fungi in an urban vegetable garden soil. Results indicated that geogenic Pb indeed modified fungi communities and altered the functional structure of collembolan communities in garden soils. Although geogenic Pb presented low bioavailability, it affected soil fauna and vegetables similar to anthropogenic Pb.     

Response of snails and slugs to fragmentation of lowland forests in NW Germany

Habitat fragmentation is a major cause for species loss, but its effect on invertebrates with low active dispersal power, like terrestrial gastropods, has rarely been studied. Such species can not cross a hostile habitat matrix, for which the predictions of island theory, such as positive relations between species richness and patch size, should apply. In order to test this prediction, we studied gastropod species diversity by assessing gastropod assemblage characteristics from 35 sites in 19 fragments of deciduous old-growth forests in the Lower Rhine Embayment, Germany. Assemblages differed between larger (≥700 ha) and smaller forests (<400 ha), those of large forests held a higher percentage of forest species. Although α-diversity was similar between the two forest size classes, small forests often comprised matrix species, resulting in a higher β-diversity. Edge effects on the species richness of matrix species were noticeable up to 250 m into the forest. Hierarchical partitioning revealed that distance to disturbances (external edge, internal edges like roads) explained most assemblage variables, whereas forest size and woodland cover within a 1 km radius from the sites explained only a few assemblage variables. Densities of two forest-associated species, Discus rotundatus and Arion fuscus, decreased with forest size. Yet, forest size was positively correlated with richness of typical forest species and densities of Limax cinereoniger. The latter species seems to need forests of >1,000 ha, i.e., well above the size of most fragments. In conclusion, the prediction is valid only for forest species. The response to fragmentation is species specific and seems to depend on habitat specialization and macroclimatic conditions. Jean-Pierre Maelfait: Deceased.     

Toll-like receptor 3 (TLR3): a new marker of canine monocytes-derived dendritic cells (cMo-DC)

Toll-like receptors (TLRs) are a family of functionally important receptors for recognition of pathogen-associated molecular pattern (PAMP) since they trigger the pro-inflammatory response and upregulation of costimulatory molecules, linking the rapid innate response to adaptative immunity. In human leukocytes, TLR3 has been found to be specifically expressed in dendritic cells (DC). This study examined the expression of TLR3 in canine monocytes-derived DC (cMo-DC) and PBMC using three new anti-TLR3 mAbs (619F7, 722E2 and 713E4 clones). The non-adherent cMo-DC generated after culture in canine IL-4 plus canine GM-CSF were labelled with the three anti-TLR3 clones by flow cytometry, with a strong expression shown for 619F7 and 722E2 clones. By contrast, TLR3 expression was low to moderate in canine monocytes and lymphocytes. These results were confirmed by Western blot using 619F7 and 722E2 clones and several polypeptide bands were observed, suggesting a possible cleavage of TLR3 molecule or different glycosylation states. In addition, TLR3 was detectable in immunocytochemistry by using 722E2 clone. In conclusion, this first approach to study canine TLR3 protein expression shows that three anti-TLR3 clones detect canine TLR3 and can be used to better characterize canine DC and the immune system of dogs.