Impact of a mutator phenotype on motility and cell adherence in Salmonella Heidelberg

In this study, we investigated adherence and motility of the hypermutator Salmonella enterica Heidelberg B182 bovine strain related to a 12bp deletion in mutS. This mutator phenotype was associated with increased adherence to epithelial cells and with high expression of fimA as shown by real-time RT-PCR. Motility studies showed that fliC were up-regulated in the B182 strain, while fljA and fljB were down-regulated. In order to determine if mutated mutS is implicated in this genes expression, isogenic strains, derived from a WT strain, containing the 12bp deletion in mutS (Δ12bpmutS) or an inactivated mutS (ΔmutS) were generated. Δ12bpmutS and ΔmutS strains showed a spontaneous mutation rate similar to the environmental strain B182, but exhibited lower adherence capacity and fimA expression. In contrast to the fimbriae genes, in Δ12bpmutS, fliC expression was up-regulated, but fljA and fljB expression were decreased, as in the B182 strain. Only fljB expression was increased in ΔmutS mutants. Taken together, our data suggest that mutS alteration does not influence fimbriae expression but can impact flagella genes.     

Predictive Factors of Sensitivity to Elisidepsin,a Novel Kahalalide F-Derived Marine Compound

Elisidepsin (PM02734, Irvalec^®) is a synthetic marine-derived cyclic peptide of the Kahalalide F family currently in phase II clinical development. Elisidepsin was shown to induce rapid oncosis in ErbB3-expressing cells. Other predictive factors of elisidepsin sensitivity remained unknown. A panel of 23 cancer cell lines of different origin was assessed for elisidepsin cytotoxicity and correlated with mutational state, mRNA and protein expression of selected genes. Elisidepsin showed potent and broad cytotoxic effects in our cancer cell line panel, being active at concentrations ranging from 0.4 to 2 μM that may be relevant for clinical settings. We have shown that elisidepsin is more active in cells harboring epithelial phenotype with high E-cadherin and low vimentin expression. In addition, high ErbB3 and Muc1 expression was correlated with sensitivity to elisidepsin, whereas the presence of KRAS activating mutations was associated with resistance. In DU-PM cells with acquired resistance to elisidepsin, ErbB3 expression was decreased, while Bcl2 was increased. DU-PM cells displayed higher sensitivity to ErbB1-inhibitors suggesting possible cross-talk of ErbB1 and ErbB3 signaling pathways. Combinations of elisidepsin with lapatinib and several chemotherapies including 5-FU and oxaliplatin resulted in synergistic effects that offer the potential of clinical use of elisidepsin in combination settings.     

Relationships between soil fauna communities and humus forms: Response to forest dynamics and solar radiation

This study investigated the responses of soil animal communities, soil functioning and humus forms to forest dynamics and solar radiation. We examined changes in invertebrate communities and soil features in two subalpine spruce forests (Eastern Italian Alps, Trento) growing on a calcareous bedrock, with different sun exposures (north and south), each forming a chronosequence of three developmental phases: clearing, regeneration stand (25-year-old trees) and mature stand (170-year-old trees). Our results indicate that the two forest sites differed in solar energy input, soil chemical properties and the relationships between forest dynamics and animal communities. In the north-facing site, soil fauna communities were very similar in the three forest developmental phases. Conversely, in the south-facing site, the composition of invertebrate communities and the diversity of zoological groups varied greatly among developmental phases. The highest abundance of total invertebrates, and mites in particular, occurred in the south-facing mature stands while the south-facing regeneration stand was characterised by higher densities of Collembola, Chilopoda, Symphyla, Protura and Aranea. The structure of communities in clearings was the same as in regeneration stands but with lower invertebrate abundance. Humus forms and soil features changed with developmental phases in both the south- and north-facing sites, although variations were more pronounced in the southern exposure. Mature stands were characterised by high levels of soil organic carbon and nitrogen, C/N values and low pH, the clearings and regeneration stands being characterised by a greater release of mineral nitrogen. The diversity of zoological groups (Shannon–Wiener index) was linearly correlated to soil pH, Humus Index, the amount of organic carbon and the species richness of herbaceous plants. Our results about the composition and the diversity of invertebrate communities are consistent with the observations of other authors studying south-exposed forests growing on different bedrock types, indicating that such relationships are widespread. The higher densities of invertebrates in the south-facing site may be attributed to higher solar radiation, and the positive correlation observed between total soil fauna abundance and solar energy supports the “more individuals” hypothesis that assumes a positive relationship between the number of individuals and energy availability. Possible ways by which forest dynamics control soil invertebrate communities are discussed.     

Comparison of bronchoalveolar lavage cytospins and smears in dogs and cats

Differences in the cytological interpretation of bronchoalveolar lavage fluid (BALF) after cytospin preparation (CP) or manual smearing of pelleted cells preparation (MSP) were investigated in client-owned dogs and cats with inflammatory or infectious lower respiratory disease. Bronchoalveolar lavage fluid from healthy cats was also examined. With MSP, cell lysis was more frequently observed, and cellular distribution was more heterogeneous throughout the slide. When samples from healthy and diseased animals were considered together, a significantly greater percentage of neutrophils was seen on CP than on MSP slides (P<0.002). Cytospin preparations were considered of better quality in all individual comparisons. Cytospin preparation is advised in the evaluation of BALF with low total cell count. When only MSPs are evaluated, clinicians should be aware that differential neutrophil counts may underestimate the counts found on CP slides.     

Responses to light in a soil-dwelling springtail

It has been widely assumed that Collembola respond to light, but until now there has been very little experimental proof of this. Field observations allowed to distinguish soil-dwelling species that would escape from light from surface-dwelling species that would be attracted to light. However, the supposed effect of light could be due to other factors such as temperature or dryness. We demonstrated that individuals of the collembolan species Heteromurus nitidus (Entomobryidae), when placed in a light gradient (temperature and moisture being homogeneous), clustered in the darker area. This effect occurred rapidly and changes in the distribution of animals persisted after illumination ceased. This shows light to act as a strong repellent for this soil-dwelling collembolan species.     

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.     

Cardinal temperatures variability within a tropical japonica rice diversity panel

Air temperature is one of the most critical climatic factors controlling rice growth, development, and production in current and future climatic scenarii predicting increasingly frequent situations of extreme and/or fluctuating temperatures. With its large spectrum of geographical origins and cropping areas, one can credit tropical japonica rice subspecies of a probable genetic diversity of its response to air temperature, which is of major interest for the breeding of better adapted rice varieties. A panel of 195 rice accessions (175 japonica plus 20 reference cultivars) was studied in controlled environment to estimate cardinal (base, optimum, and maximum) temperatures based on the monitoring of the elongation rate (LERmax) of the sixth leaf on the main stem in response to six fixed thermal treatments ranging from 16 to 35 °C. A dedicated statistical framework was elaborated for estimating LERmax, cardinal temperature and related uncertainties. Developed statistical framework enhanced the precision of cardinal temperatures estimated compared to previously reported methods, especially for base temperature. Maximum temperature was trickier to estimate and will require further studies. A significant genotypic variability for base and optimal temperature was pointed out, suggesting tropical japonica subspecies represents a relevant genetic pool to breed for rice genotypes adapted to various thermal situations. These results also suggested that using genotype-dependent cardinal temperature values should enhance the way crop growth models account for genotype?×?environment interactions hence their predictive value in current and future climatic conditions.     

Immunological responses following experimental endobronchial infection of badgers (Meles meles) with different doses of Mycobacterium bovis

The Eurasian badger (Meles meles) is a wildlife reservoir for Mycobacterium bovis infection in Ireland and Great Britain and has been implicated in the transmission of tuberculosis to cattle. Vaccination of badgers is an option that could be used as part of a strategy to control the disease. In this study we used an endobronchial infection procedure to inoculate groups of badgers with three different doses (3x10(3), 2x10(2) and <10 Colony Forming Units (CFUs)) of M. bovis. After 17 weeks the disease status of each animal was determined by post-mortem pathology and culture for M. bovis. Each of the inoculum doses resulted in establishment of infection in the badgers. The cell-mediated immune (CMI) responses were measured by lymphocyte transformation assay (LTA) of peripheral blood mononuclear cells (PBMCs) cultured with bovine tuberculin (PPD-B). In each infected group the CMI responses increased with a kinetic profile corresponding to the delivered dose and the post-mortem pathology. The serological responses were measured by ELISA and a multi-antigen print immunoassay (MAPIA) in order to investigate any changes in the antigenic repertoire associated with different infective doses. In contrast to the CMI responses, the ELISA and MAPIA showed that the recognition of antigens by the badgers was intermittent and not strongly influenced by the dose of M. bovis.     

Canine visceral leishmaniasis: comparison of in vitro leishmanicidal activity of marbofloxacin, meglumine antimoniate and sodium stibogluconate

The control of canine leishmaniasis largely depends on the success of treatment. Drugs currently available to treat this disease are toxic and partially effective. The curative effect of marbofloxacin, a third-generation fluoroquinolone developed for veterinarian individual treatment, was evaluated in vitro in the presence of Leishmania infantum promastigotes and dog-monocyte-derived macrophages; meglumine antimoniate and sodium stibogluconate were used as comparative treatments. We observed that the killing of Leishmania promastigotes and intracellular amastigotes by marbofloxacin was dose-dependent. We demonstrated that successful treatment of canine infected macrophages for 48 h was possible with 500 microg/ml of marbofloxacin. Leishmanicidal activity acted through a TNF-alpha and nitric oxide pathway and correlated with the generation of nitric oxide (NO(2)) production by monocytes derived macrophages from infected (23+/-5 microM) or healthy (21+/-6 microM) dogs, in comparison with NO(2) concentration in infected/non-treated macrophages (< 3 microM, P<0.01). This significant induced parasiticidal effect correlated with extensive elimination of amastigotes by macrophages derived from infected (11+/-5) and healthy dogs (6+/-2), when compared to infected/non-treated macrophages (530+/-105 and 472+/-86 amastigotes, respectively, P< 0.01). Marbofloxacin was shown to be non-toxic at 500 microg/ml in vitro and no cell apoptosis was observed. The molecule was able to induce a parasitic process after significant elimination of amastigotes in leishmania-infected dog macrophages. We propose that marbofloxacin, compared to standard chemotherapeutic agents (meglumine antimoniate and sodium stibogluconate), could be an effective and pragmatic oral route alternative to treat canine leishmaniasis.