Isolation and Characterization of a Novel Arabidopsis thaliana Mutant Unable to Develop Wilt Symptoms After Inoculation with a Virulent Strain of Ralstonia solanacearum

ABSTRACT To characterize host genes required for a compatible interaction, we identified a novel recessive Arabidopsis thaliana mutant, nws1 (no wilt symptoms), that failed to develop wilt symptoms in response to virulent strains of the phytopathogenic bacterium, Ralstonia solanacearum. The absence of wilting in nws1 plants was not correlated with a cell death phenotype or a constitutive expression of salicylic acid-, jasmonic acid- or ethylene-associated genes. In addition, this mutation, which conferred a symptomless phenotype in response to all the R. solanacearum strains tested, was highly specific to this pathogen, because nws1 responses to other plant pathogens, including oomycetes, nematodes, viruses, and other bacteria, were identical to those of wild-type Col-5 plants. Finally, the lack of disease development was shown to be different than RRS1-R-mediated resistance. The identification of mutants such as nws1, that are unable to develop disease, should lead to the isolation of target host factors required for pathogen growth or fitness, or of factors modified by the invading microorganism to avoid or inactivate plant defense mechanisms, and should bring a better understanding of bacterial wilt diseases.     

Oral immunisation of pigs with fimbrial antigens of enterotoxigenic E. coli: an interesting model to study mucosal immune mechanisms

The intestinal mucosal immune system can discriminate actively between harmful pathogenic agents and harmless food antigens resulting in different immune responses namely IgA production and oral tolerance, respectively. Recently, a pig model has been developed for studying intestinal mucosal immune responses in which F4 fimbrial antigens of enterotoxigenic Escherichia coli (F4 ETEC) are used as oral antigens. A unique feature of this model is that soluble F4 antigens can be administered to pigs which have a receptor for this fimbriae (F4R(+)) on their small intestinal villous enterocytes and pigs which do not have this receptor (F4R(-)). Oral administration of F4 to the F4R(+) pigs results in an intestinal mucosal immune response that completely protects the pigs against a challenge infection. In F4R(-) pigs such an intestinal mucosal immune response does not occur. However, a priming of the systemic immune system can be seen similar to the priming in pigs fed with the same dose of a food antigen, suggesting that F4 in F4R(-) pigs behaves as a food antigen. The fact that different mucosal immune responses can be induced with soluble F4, makes it an interesting model to study mucosal immune mechanisms in the pig.     

Control of gastrointestinal parasitism with nematodes in dairy goats by treating the host category at risk

Infections of the gastrointestinal tract with parasitic nematodes remain one of the main limiting factors in grazing dairy goats. The usual mode of control of these parasitic diseases has up to now been based on the repeated use of anthelmintics. However, the prevalence rates of anthelmintic resistances, in particular to benzimidazoles, are now particularly high in the French dairy goat production. This situation makes it mandatory to reconsider the usual mode of control of these nematodes and to look for short term, alternative solutions which combine the control of gastrointestinal infections and management of anthelmintic resistances. One of the possible options is to leave a part of the flock without treatment during the grazing season in order to maintain alleles of susceptibility to anthelmintics within the worm populations. Previous epidemiological observations identifying the categories of host populations at risk are presented which provide the rationale for targeted applications of treatments. The results of assays on experimental flocks and from farm surveys examining the advantages and drawbacks of selective treatments are presented. The value of these results in combination with other alternative solutions of control are discussed in order to use minimum treatments with maximum benefits.     

Monitoring of the ventilatory status of anesthetized birds of prey by using end-tidal carbon dioxide measured with a microstream capnometer.

The relationship between end-tidal partial pressure of carbon dioxide (PETCO2), arterial partial pressure of carbon dioxide (PaCO2), and blood pH in isoflurane-anesthetized raptors was evaluated. PaCO2 and pH were determined in serial arterial samples from isoflurane anesthetized birds and compared with concurrent end-tidal partial pressure of carbon dioxide measured with a Microstream sidestream capnograph. Forty-eight paired samples, taken from 11 birds of prey (weighing 416-2,062 g), were used to determine correlations coefficients between PaCO2 and PETCO2, and between PETCO2 and pH. Limits of agreement between PaCO2 and PETCO2 also were calculated. Strong correlations were observed between PaCO2 and PETCO2 (r = 0.94; P < 0.0001) as well as between PETCO2 and pH (r = -0.90; P < 0.0001). However, the level of agreement between PaCO2 and PETCO2 varied considerably. Low values of PETCO2, ranging from 18 to 29 mm Hg, exceeded the concomitantly measured values of PaCO2 by an average of 6.0 mm Hg (6.0 +/- 1.9 mm Hg; mean +/- SD). Conversely, high values of PETCO2, ranging from 50 to 63 mm Hg, were on average 7.6 mm Hg (7.6 +/- 9.8 mm Hg) lower than values of PaCO2. In the 30 to 49 mm Hg range for PETCO2, the difference between PETCO2 and PaCO2 was on average 1.0 mm Hg (1.0 +/- 8.5 mm Hg). These results suggest that the capnograph used provided a sufficiently accurate estimation of arterial partial pressure of carbon dioxide for birds weighing > 400 g and receiving manual positive ventilation with a Bain system. In our study, the linear relationship observed between the pH and the end-tidal partial pressure of carbon dioxide suggested that the monitoring of end-tidal partial pressure of carbon dioxide also can be useful to prevent respiratory acidosis.     

Modeling recent bark stripping by red deer (Cervus elaphus) in South Belgium coniferous stands

? Context

Over the past few decades, the impact of large herbivorous ungulates on forest vegetation has been clearly highlighted. Among those impacts, bark stripping of coniferous trees is one of the most damaging. Bark stripping leads to rot development, inducing serious loss of timber value.

? Aims

The present study aimed firstly at evidencing the factors explaining the variations observed in fresh bark peeling rate for spruce and Douglas-fir in southern Belgium and secondly at identifying the key factors to consider when setting up a deer management plan.

? Method

Fresh bark peeling rate was recorded with a systematic sampling survey from 2004 to 2007. The covered territory was then divided into 63 distinct hunting zones of area ranging from 1,000 to 25,000 ha. About 5,000 plots were monitored annually. Each zone was characterized with a large number of explanatory variables. The explanatory variables were integrated firstly into fixed linear models using a stepwise procedure, and then into a mixed model.

? Results

The significant variables included in the model (R ²?=?44 %) are (by decreasing order of importance) red deer densities, proportion of coniferous stands and agricultural areas, snow cover, distance to urban habitats, and species diversity in the understory.

? Conclusion

The models revealed the impacts of several factors on bark peeling: deer density, deer-carrying capacity of the territory, landscape structure, and severity of winter conditions. The adjusted model allowed subtracting the impact of winter conditions in order to produce a relevant indicator for hunting management. In addition, the model was used to assess the sensitivity of a forested area to bark peeling based on its environmental characteristics.     

Early infections by myxoma virus of young rabbits (Oryctolagus cuniculus) protected by maternal antibodies activate their immune system and enhance herd immunity in wild populations

The role of maternal antibodies is to protect newborns against acute early infection by pathogens. This can be achieved either by preventing any infection or by allowing attenuated infections associated with activation of the immune system, the two strategies being based on different cost/benefit ratios. We carried out an epidemiological survey of myxomatosis, which is a highly lethal infectious disease, in two distant wild populations of rabbits to describe the epidemiological pattern of the disease. Detection of specific IgM and IgG enabled us to describe the pattern of immunity. We show that maternal immunity attenuates early infection of juveniles and enables activation of their immune system. This mechanism associated with steady circulation of the myxoma virus in both populations, which induces frequent reinfections of immune rabbits, leads to the maintenance of high immunity levels within populations. Thus, myxomatosis has a low impact, with most infections being asymptomatic. This work shows that infection of young rabbits protected by maternal antibodies induces attenuated disease and activates their immune system. This may play a major role in reducing the impact of a highly lethal disease when ecological conditions enable permanent circulation of the pathogen.     

A new approach to ecological land classification for the Canadian boreal forest that integrates disturbances

Traditional approaches to ecological land classification (ELC) can be enhanced by integrating, a priori, data describing disturbances (natural and human), in addition to the usual vegetation, climate, and physical environment data. To develop this new ELC model, we studied an area of about 175,000 km² in the Abies balsamea–Betula papyrifera and Picea mariana-feathermoss bioclimatic domains of the boreal forest of Québec, in eastern Canada. Forest inventory plots and maps produced by the Ministère des Ressources naturelles du Québec from 1970 to 2000 were used to characterize 606 ecological districts (average area 200 km²) according to three vegetation themes (tree species, forest types, and potential vegetation-successional stages) and four sets of explanatory variables (climate, physical environment, natural and human disturbances). Redundancy, cluster (K-means) and variation partitioning analyses were used to delineate, describe, and compare homogeneous vegetation landscapes. The resulting ELC is hierarchical with three levels of observation. Among the 14 homogeneous landscapes composing the most detailed level, some are dominated by relatively young forests originating from fires dating back to the period centered on 1921. In others, forest stands are older (fires from the period centered on 1851), some are under the influence of insect outbreaks and fires (southern part), while the rest are strongly affected by human activities and Populus tremuloides expansion. For all the study area and for parts of it, partitioning reveals that natural disturbance is the dominant data set explaining spatial variation in vegetation. However, the combination of natural disturbances, climate, physical environment and human disturbances always explains a high proportion of variation. Our approach, called “ecological land classification of homogeneous vegetation landscapes”, is more comprehensive than previous ELCs in that it combines the concepts and goals of both landscape ecology and ecosystem-based management.     

Comparison of respiratory function during TIVA and isoflurane anaesthesia in ponies Part II: breathing patterns and transdiaphragmatic pressure

ObjectiveTo compare breathing patterns and transdiaphragmatic pressure during total intravenous (TIVA) and isoflurane anaesthesia in ponies.Study designExperimental, cross–over study.AnimalsSix healthy ponies weighing 286 (233–388) ± 61 kg, age 13 (9–16) ± 3 years.MethodsFollowing premedication with romifidine [80 μg kg^?1 intravenously (IV)], general anaesthesia was induced with midazolam (0.06 mg kg^?1 IV) and ketamine (2.5 mg kg^?1 IV) and maintained with either isoflurane (Fe’Iso = 1.1%) (T-ISO) or an IV combination of romifidine (120 μg kg^?1 per hour), midazolam (0.09 mg kg^?1 hour^?1) and ketamine (3.3 mg kg^?1 hour^?1) (T-TIVA), while breathing 60% oxygen (FIO₂). The circumference changes of the rib cage (RC) and abdominal compartment (ABD) were recorded using respiratory ultrasonic plethysmography (RUP). Balloon tipped catheters were placed in the distal oesophagus and the stomach and maximal transdiaphragmatic pressure (P_{di max)} was calculated during Mueller's manoeuvre.ResultsThe breathing pattern T-ISO was more regular and respiratory rate significantly lower compared with T-TIVA. Ponies in T-TIVA showed regularly appearing sighs, which were never observed in T-ISO. Different contribution of the RC and ABD compartments to the breathing pattern was observed with a smaller participation of the RC to the total volume change during T-ISO. Transdiaphragmatic pressures (mean 13.7 ± SD 8.61 versus 23.4 ± 7.27 cmH₂O, p < 0.0001) were higher in T-TIVA compared to T-ISO. The sum of the RC and ABD circumferential changes was lower during T-TIVA compared to T-ISO (6.32 ± 4.42 versus 11.72 ± 4.38 units, p < 0.0001).Conclusion and clinical relevanceMarked differences in breathing pattern and transdiaphragmatic pressure exist during inhalation- and TIVA and these should be taken into account for clinical estimation of anaesthetic depth.     

Variety of non‐invasive continuous monitoring methodologies including electrical impedance tomography provides novel insights into the physiology of lung collapse and recruitment – case report of an anaesthetized horse

IntroductionThe use of alveolar recruitment maneuvers during general anaesthesia of horses is a potentially useful therapeutic option for the ventilatory management. While the routine application of recruitments would benefit from the availability of dedicated large animal ventilators their impact on ventilation and perfusion in the horse is not yet well documented nor completely understood.Case historyA healthy 533 kg experimental horse underwent general anaesthesia in lateral recumbency. During intermittent positive pressure ventilation a stepwise alveolar recruitment maneuver was performed.ManagementAnaesthesia was induced with ketamine and midazolam and maintained with isoflurane in oxygen using a large animal circle system. Mechanical ventilation was applied in pressure ventilation mode and an alveolar recruitment maneuver performed employing a sequence of ascending and descending positive end expiratory pressures. Next to the standard monitoring, which included spirometry, additionally three non-invasive monitoring techniques were used: electrical impedance tomography (EIT), volumetric capnography and respiratory ultrasonic plethysmography. The functional images continuously delivered by EIT initially showed markedly reduced ventilation in the dependent lung and allowed on-line monitoring of the dynamic changes in the distribution of ventilation during the recruitment maneuver. Furthermore, continuous monitoring of compliance, dead space fraction, tidal volumes and changes in end expiratory lung volume were possible without technical difficulties.Follow upThe horse made an unremarkable recovery.ConclusionThe novel non-invasive monitoring technologies used in this study provided unprecedented insights into the physiology of lung collapse and recruitment. The synergic information of these techniques holds promise to be useful when developing and evaluating new ventilatory strategies in horses.     

Fluorescent tagging of VP22 in N-terminus reveals that VP22 favors Marek’s disease virus (MDV) virulence in chickens and allows morphogenesis study in MD tumor cells

Marek’s disease virus (MDV) is an alpha-herpesvirus causing Marek’s disease in chickens, mostly associated with T-cell lymphoma. VP22 is a tegument protein abundantly expressed in cells during the lytic cycle, which is essential for MDV spread in culture. Our aim was to generate a pathogenic MDV expressing a green fluorescent protein (EGFP) fused to the N-terminus of VP22 to better decipher the role of VP22 in vivo and monitor MDV morphogenesis in tumors cells. In culture, rRB-1B EGFP22 led to 1.6-fold smaller plaques than the parental virus. In chickens, the rRB-1B EGFP22 virus was impaired in its ability to induce lymphoma and to spread in contact birds. The MDV genome copy number in blood and feathers during the time course of infection indicated that rRB-1B EGFP22 reached its two major target cells, but had a growth defect in these two tissues. Therefore, the integrity of VP22 is critical for an efficient replication in vivo, for tumor formation and horizontal transmission. An examination of EGFP fluorescence in rRB-1B EGFP22-induced tumors showed that about 0.1% of the cells were in lytic phase. EGFP-positive tumor cells were selected by cytometry and analyzed for MDV morphogenesis by transmission electron microscopy. Only few particles were present per cell, and all types of virions (except mature enveloped virions) were detected unequivocally inside tumor lymphoid cells. These results indicate that MDV morphogenesis in tumor cells is more similar to the morphorgenesis in fibroblastic cells in culture, albeit poorly efficient, than in feather follicle epithelial cells.