Partial divergence of cytokine mRNA expression in bronchial tissues compared to bronchoalveolar lavage cells in horses with recurrent airway obstruction

The aim of this study was to investigate mRNA levels of cytokines in bronchial epithelium in horses with recurrent airway obstruction (RAO) during acute crisis and remission. Additionally, cytokine mRNA levels in endobronchial biopsies and bronchoalveolar lavage (BAL) cells were compared. Seven RAO horses were examined while in respiratory crisis following provocation and again while in remission after 2 months on pasture, during which time six healthy horses on pasture were also examined. Quantitative real-time PCR (RT-PCR) was used to assess mRNA expression for cytokines IL-5, IL-6, IL-8, IL-10, IL-17 and transforming growth factor beta1 (TGF-beta1) in endobronchial biopsies and bronchoalveolar lavage. Expression of IL-8 mRNA was significantly upregulated during crisis in both endobronchial biopsies and BAL cells (p=0.036), while there was a similar trend for upregulation of IL-10 mRNA only in BAL cells that approached significance (p=0.059). Moreover, during crisis the expression of IL-8 mRNA in BAL cells was positively correlated to relative IL-6 mRNA expression (r(s)=0.971, p=0.001) and bronchial epithelial expression of IL-10 and TGF-beta1 mRNA were positively correlated (r(s)=0.943, p=0.005). In comparing the relationship of mRNA expression in BAL to biopsy in individual RAO horses, there was a positive correlation with IL-6 to IL-8 mRNA expression in BAL during respiratory crisis (r(s)=0.971, p=0.001) that also correlated positively with IL-8 expression in biopsies on pasture (r(s)=0.986, p<0.0001 for both). Regarding RAO horses at pasture versus controls neither the cytokine mRNA levels in endobronchial biopsy nor in BAL cells differed significantly. These results further support previous findings that IL-8 mRNA in both BAL cells and bronchial epithelium is upregulated in RAO horses during crisis. However, apart from IL-8, it appears that expression of other cytokines, including IL-5, IL-6, IL-10, IL-17 and TGF-beta1 in bronchial epithelium does not necessarily mirror cytokine expression in BAL cells in individual horses with RAO. Accordingly, examination of markers of inflammation in endobronchial tissue provides complementary but not necessarily identical information to that obtained in BAL cells. Given the potential for repeated sampling over time bronchial biopsy can serve as an invaluable additional tool for investigation of time-dependent changes in inflammatory process in this animal model of asthma.     

Temporal shifts in landscape connectivity for an ecosystem engineer,the roe deer,across a multiple-use landscape

Context

Routine movements of large herbivores, often considered as ecosystem engineers, impact key ecological processes. Functional landscape connectivity for such species influences the spatial distribution of associated ecological services and disservices.

Objectives

We studied how spatio-temporal variation in the risk-resource trade-off, generated by fluctuations in human activities and environmental conditions, influences the routine movements of roe deer across a heterogeneous landscape, generating shifts in functional connectivity at daily and seasonal time scales.

Methods

We used GPS locations of 172 adult roe deer and step selection functions to infer landscape connectivity. In particular, we assessed the influence of six habitat features on fine scale movements across four biological seasons and three daily periods, based on variations in the risk-resource trade-off.

Results

The influence of habitat features on roe deer movements was strongly dependent on proximity to refuge habitat, i.e. woodlands. Roe deer confined their movements to safe habitats during daytime and during the hunting season, when human activity is high. However, they exploited exposed open habitats more freely during night-time. Consequently, we observed marked temporal shifts in landscape connectivity, which was highest at night in summer and lowest during daytime in autumn. In particular, the onset of the autumn hunting season induced an abrupt decrease in landscape connectivity.

Conclusions

Human disturbance had a strong impact on roe deer movements, generating pronounced spatio-temporal variation in landscape connectivity. However, high connectivity at night across all seasons implies that Europe’s most abundant and widespread large herbivore potentially plays a key role in transporting ticks, seeds and nutrients among habitats.

     

Deuterobiosphere the Chemosynthetic Second Biosphere of the Globe. A First Review

With the discovery of Ophel, a chemosynthesis‐based continental subterranean environment (Por 2007), it became evident that such environments, previously known only from the sea, are spanning the world as a parallel biosphere, the deuterobiosphere. In this preliminary contribution the different known biomes of the deuterobiosphere are reviewed and some general and comparative features are listed.     

Finite Mixture of Regression Modeling for High-Dimensional Count and Biomass Data in Ecology

Understanding how species distributions respond as a function of environmental gradients is a key question in ecology, and will benefit from a multi-species approach. Multi-species data are often high dimensional, in that the number of species sampled is often large relative to the number of sites, and are commonly quantified as either presence–absence, counts of individuals, or biomass of each species. In this paper, we propose a novel approach to the analysis of multi-species data when the goal is to understand how each species responds to their environment. We use a finite mixture of regression models, grouping species into “Archetypes” according to their environmental response, thereby significantly reducing the dimension of the regression model. Previous research introduced such Species Archetype Models (SAMs), but only for binary assemblage data. Here, we extend this basic framework with three key innovations: (1) the method is expanded to handle count and biomass data, (2) we propose grouping on the slope coefficients only, whilst the intercept terms and nuisance parameters remain species-specific, and (3) we develop model diagnostic tools for SAMs. By grouping on environmental responses only, the model allows for inter-species variation in terms of overall prevalence and abundance. The application of our expanded SAM framework data is illustrated on marine survey data and through simulation. Supplementary materials accompanying this paper appear on-line.     

Widespread distribution of resistance to triazole fungicides in Brazilian populations of the wheat blast pathogen

Fungicides have not been effective in controlling the wheat blast disease in Brazil. An earlier analysis of 179 isolates of Pyricularia oryzae Triticum lineage (PoTl) sampled from wheat fields across six populations in central-southern Brazil during 2012 discovered a high level of resistance to strobilurin fungicides. Here we analysed azole resistance in the same strains based on EC₅₀ measurements for tebuconazole and epoxiconazole. All six Brazilian populations of PoTl exhibited high resistance to both azoles, with in vitro EC₅₀ values that were at least 35 to 50 times higher than the recommended field doses. We sequenced the CYP51A and CYP51B genes to determine if they were likely to play a role in the observed azole resistance. Although we found five distinct haplotypes in PoTl carrying four nonsynonymous substitutions in CYP51A, none of these substitutions were correlated with elevated EC₅₀. CYP51B was sequenced for nine PoTl isolates, three each representing low, medium, and high tebuconazole EC₅₀. Both PoTl CYP51A and CYP51B could complement yeast CYP51 function. All PoTl CYP51A-expressing yeast transformants were less sensitive to triazoles than the PoTl CYP51B ones. Transformants expressing PoTl CYP51A haplotype H1 carrying the R158K substitution were not more resistant than those expressing PoTl CYP51A haplotype H5, which is synonymous to haplotype H6, found in triazole-sensitive P. oryzae Oryza isolates from rice blast. Therefore, the reduced triazole sensitivity of wheat blast isolates compared to rice blast isolates appears to be associated with a non-target-site related resistance mechanism acquired after higher exposure to triazoles.     

The influence of oral bacteria on tissue levels of Toll-like receptor and cytokine mRNAs in feline chronic gingivostomatitis and oral health

Feline chronic gingivostomatitis (FCGS) is an inflammatory disease of the oral cavity that causes severe pain and distress in affected cats. Treatment methods are currently very limited. The aims of this study were to assess the feline innate immune response by investigating the levels of cytokine and Toll-like receptor (TLR) mRNAs in tissue biopsies of cats with and without FCGS, and to relate this to the presence or absence of putative oral pathogens identified previously within these cats. Mucosal biopsies were collected from 28 cats with FCGS and eight healthy cats. The levels of TLR (TLR2, TLR3, TLR4, TLR7, TLR9) and cytokine (IL-1β, IL-4, IL-6, IL-10, IL-12, TNF-α, IFN-γ) mRNA was determined using quantitative PCR. In the FCGS group a statistically significant increase was seen in TLR2, TLR7, TNF-α, IFN-γ, IL-1β and IL-6 mRNA levels compared to the healthy group. In cats where Tannerella forsythia was present, statistically significant increases were seen in TLR2, TLR4, TLR7, TLR9, TNF-α and IL-1β mRNA levels compared to cats where this putative pathogen was absent. Statistically significant increases in mRNA expression were also seen in cats harbouring feline calicivirus (FCV) (TLR2, IL-1β, IL-6, IFN-γ) and Porphyromonas circumdentaria (TLR2, TLR3) compared to cats where these putative pathogens were absent. Pasteurella multocida subsp. multocida and Pseudomonas sp. did not significantly alter the expression of any TLR or cytokine mRNAs when compared to animals who tested negative for these species, while cats colonised with P. multocida subsp. septica demonstrated a statistically significant reduction in the expression of TLR7, TNF-α and IFN-γ mRNAs compared to cats free of this species. The expression of mRNA for several TLRs and cytokines is elevated in FCGS. A positive correlation was observed between clinical disease severity and the presence of FCV (p = 0.001; Rho = 0.58). Although the number of cats harbouring T. forsythia was low by comparison, 80% of samples in which it was present were from cases with the highest clinical disease severity. Positive correlations with clinical disease severity were seen for TLR2 (p = 0.00086), TLR7 (p = 0.049), TNF-α (p = 0.027), IFN-γ (p = 0.0015), IL-1β (p = 0.004) and IL-6 (p = 0.00001) mRNAs. The putative pathogens FCV and T. forsythia may be important in stimulating a host immune response to FCGS and may play a role in the pathogenesis of this disease.     

Soil physico-chemical changes following application of municipal solid waste leachates to grasslands

Concerns about the use of residues from municipal solid waste incinerators (MSWI) in construction materials usually focus on the potential for heavy metals and organic chemicals to leach into drainage waters under the influence of rain. We hypothesised that high level of salts in the MSWI leachates may cause more of a problem, particularly on soil physico-chemical properties. Both bottom ash (BA) and Solidified Air Pollution Control residue (SAPCr) leachates were added to experimental grassland plots. The amounts of Na⁺ increased by up to 13% in soils supplemented with each leachate. A decrease of the soil total porosity (?14%) was evidence of a subsequent adverse physical effect of this strong salinity. The potential for the grass cover type (species composition or density) to limit this adverse effect was discussed. Laboratory tests allowed us to determine that undiluted SAPCr induced slaking of aggregates accompanied by a strong decrease of aggregate stability, to 49% of control values. Undiluted BA induced dispersion of clays and others fine particles, which are then dislodged and transported into pores, causing blockage and decreasing total porosity. Clay dispersion followed by aggregate collapse occurred when soil solution contaminated by SAPCr was diluted by rainwater. This work stressed the importance of accounting for mineral contaminants, such as salts, when conducting an assessment of waste reuse scenarios.     

Reproduction of postweaning multisystemic wasting syndrome in pigs experimentally inoculated with a Swedish porcine circovirus 2 isolate.

In recent years, porcine circovirus type 2 (PCV2)-associated postweaning multisystemic wasting syndrome (PMWS) has been reported worldwide. However, to date, PMWS has not been reported in Sweden despite the demonstration of serum antibodies to a PCV2-like virus in Swedish pigs. This communication reports the experimental reproduction of clinical PMWS after inoculation of colostrum-deprived (CD) pigs, derived from a Northern Ireland herd, with an isolate of PCV2 virus recovered from a clinically normal Swedish pig that was necropsied in 1993. The clinical disease and histological lesions observed in CD pigs inoculated with this virus were indistinguishable from those observed in previous studies on CD pigs inoculated with a PCV2 virus isolate recovered from pigs with PMWS. These results highlight the disease potential of PCV2 isolated from regions apparently free of PMWS and suggest that the status of the host and its environment is an important factor in the development of clinical PMWS.     

Electrocardiographic findings in miniature schnauzers with syncope

Objective: The purpose of this study was to examine a population of miniature schnauzers exhibiting syncope as their primary presenting complaint and to correlate their electrocardiographic (ECG) findings with the criteria for the diagnosis of sick sinus syndrome (SSS) reported in the human and veterinary literature. Design: Retrospective study. Patients: Case records of 31 miniature schnauzers with syncope during the period 1996–2000 were reviewed. Measurements and results: This study supported two previous studies finding that females were more predisposed to ECG abnormalities associated with SSS than males. It was also noted that females were slightly older (10.1 years) when compared to males (8.9 years) at the time of diagnosis. A heart murmur was present in 61% of the dogs and this was most likely secondary to mitral valve endocardiosis and regurgitation. The primary rhythm disturbance seen was sinus arrest/sinoatrial block in 48.5% of the cases; sinus bradycardia was the second most common arrhythmia noted. Conclusion: In miniature schnauzers with syncope, SSS should be considered as a primary etiology if abnormal ECG findings are noted.