Screening of yeasts as probiotic based on capacities to colonize the gastrointestinal tract and to protect against enteropathogen challenge in mice

Probiotics are defined as viable microorganisms that exhibit a beneficial effect on the host's health when they are ingested. Two important criteria are used for selection of probiotic microorganisms: they must be able to survive in the gastrointestinal environment and to present at least one beneficial function (colonization resistance, immunomodulation or nutritional contribution). Generally, in vitro assays demonstrating these properties were used to select probiotics but it is unclear if the data can be extrapolated to in vivo conditions. In the present work, twelve Saccharomyces cerevisiae strains isolated from different environments (insect association, tropical fruit, cheese and "aguardente" production) and pre-selected for in vitro resistance to simulated gastrointestinal conditions were inoculated in germ-free mice to evaluate their real capacity to colonize the mammal digestive tract. Using these data, one of the yeasts (S. cerevisiae 905) was selected and tested in gnotobiotic (GN) and conventional (CV) mice for its capacity to protect against oral challenge with two enteropathogenic bacteria (Salmonella Typhimurium and Clostridium difficile). The yeast reached populational levels potentially functional in the gastrointestinal portions where the enteropathogens tested act. No antagonism against either pathogenic bacterium by the yeast was observed in the digestive tract of GN mice but, after challenge with S. Typhimurium, mortality was lower and liver tissue was better preserved in CV animals treated with the yeast when compared with a control group (p<0.05). Histopathological results of intestines showed that the yeast also presented a good protective effect against oral challenge with C. difficile in GN mice (p<0.05). In conclusion, among the 12 S. cerevisiae tested, strain 905 showed the best characteristics to be used as a probiotic as demonstrated by survival capacity in the gastrointestinal tract and protective effect of animals during experimental infections.     

Trypanosoma cruzi: influence of predominant bacteria from indigenous digestive microbiota on experimental infection in mice

To verify the influence of some predominant components from indigenous microbiota on systemic immunological responses during experimental Chagas disease, germ-free NIH Swiss mice were mono-associated with Escherichia coli, Enterococcus faecalis, Bacteroides vulgatus or Peptostreptococcus sp. and then infected with the Y strain of Trypanosoma cruzi. All the mono-associations predominantly induced a Th1 type of specific immune response to the infection by T. cruzi. A direct correlation was observed between a higher survival rate and increased IFN-gamma and TNF-alpha production (P<0.05) in E. faecalis-, B. vulgatus-, and Peptostreptococcus-associated mice. Moreover, higher levels of anti-T. cruzi IgG1 and anti-T. cruzi IgG2a were also found in mono-associated animals after infection. On the other hand, with the exception of E. faecalis-associated mice, mono-association induced a lower IL-10 production after infection (P<0.05) when compared with germ-free animals. Interestingly, spleen cell cultures from non-infected germ-free and mono-associated mice spontaneously produced higher levels (P<0.05) of IL-10 than cultures from infected mono-associated mice, except again for E. faecalis-associated animals. In conclusion, the presence of the components of the indigenous microbiota skews the immune response towards production of inflammatory cytokines during experimental infection with T. cruzi in gnotobiotic mice. However, the degree of increase in production of cytokines depends on each bacterial component.     

Activity following arousal in winter in North American vespertilionid bats

• 1 Many bat species hibernate to conserve energy during winter and like all hibernators they commonly arouse. During these arousals, some bats may undertake activities away from the hibernation site. Systematic ecological studies of the frequency and purpose of winter activity in temperate zone bats of North America are rare and much of the literature involves observations of single individuals or unmarked populations.
• 2 We review the available literature on winter activity among North American vespertilionid bats to highlight the paucity of data on this subject and to stimulate future research. Due to the lack of repeated, systematic studies on most North American species, the conclusions drawn are general or pertain only to parts of the geographical range of any species.
• 3 We suggest that winter activity is ubiquitous among temperate zone bats, but the degree and purpose of activity varies greatly between and within species. In general, cave‐dwelling bats tend to be relatively inactive compared with tree and foliage roosting bats during winter.
• 4 Cave‐dwelling and tree cavity‐dwelling species do not appear to feed, but they do drink and occasionally copulate during the hibernation period. Species that hibernate in foliage or leaf litter are the most active species during winter and may feed and drink throughout winter, although they probably do not copulate because of their solitary nature.
• 5 We encourage researchers to conduct studies on all aspects of winter activity for North American vespertilionids across wide latitudinal and altitudinal gradients. We suggest studies in the near future be focused on estimating the energetic costs and benefits of winter activity through determination of the frequency and intensity of winter feeding activity and more thorough examinations of movements within and among caves. Studies on common, wide‐ranging species have the potential to illuminate large‐scale patterns and differences and should be favoured over studies on rarer species.

     

Detection of Helicobacter Species in the Gastrointestinal Tract of Wild Rodents From Brazil

Since we have limited knowledge about the occurrence of Helicobacter in wild animals, we searched for Helicobacter species in the gastrointestinal tract of 75 rodents captured in forest remnants of Minas Gerais, Brazil. Fragments from the antrum and corpus of the stomach and from the colon were taken for PCR assays for Helicobacter detection. Although gastric mucosa was Helicobacter-positive in only one animal, the bacterium was detected in the colonic mucosa of 23 rodents (30.7%). Helicobacter detection was more frequent in the colon of terraced rice rat (56%) and house rat (30%) in contrast to punare and Spix’s yellow-toothed cavy, in which the presence of the bacterium was not detected. Helicobacter rodentium, H. marmotae, H. cinaedi, and other species closely related to the murine helicobacters were presumptively identified by DNA sequencing. Wild rodents may serve as a reservoir of these Helicobacter species in nature.     

Comparison of antagonistic ability against enteropathogens by G+ and G− anaerobic dominant components of human fecal microbiota

To confirm if anaerobic G+-components are those responsible for the function of colonization resistance, obligate anaerobic G+- and G- -bacteria from normal dominant microbiota of human feces were isolated from three successive collections and then used in in vitro assays for antagonism against two enteropathogenic bacteria. The production of inhibitory diffusible compounds was determined on supplemented BHI agar and MRS agar media for G- - and G+-bacteria, respectively. Salmonella enterica subsp. enterica serovar Typhimurium and Shigella sonnei were used as indicators. G+-bacteria presented a higher overall antagonistic frequency against both pathogenic bacteria (57 and 64 % for S. enterica serovar Typhimurium and S. sonnei, respectively) when compared to G+-microorganisms but with a quite elevated variation between volunteers (0-100 %) and collection samples (40-72 and 40-80 % for S. enterica sv. Typhimurium and S. sonnei, respectively). On the other hand, only three among 143 G- -isolates tested showed antagonistic activity. The results showed that, at least in vitro, obligate anaerobic G+-components of the dominant human fecal microbiota present a higher potential for antagonism against the enteropathogenic models tested than do G- -bacteria.     

Compression or tension? The stress distribution in the proximal femur

Background  

Questions regarding the distribution of stress in the proximal human femur have never been adequately resolved. Traditionally, by considering the femur in isolation, it has been believed that the effect of body weight on the projecting neck and head places the superior aspect of the neck in tension. A minority view has proposed that this region is in compression because of muscular forces pulling the femur into the pelvis. Little has been done to study stress distributions in the proximal femur. We hypothesise that under physiological loading the majority of the proximal femur is in compression and that the internal trabecular structure functions as an arch, transferring compressive stresses to the femoral shaft.     

Comparative adaptations of Aphanizomenon and Anabaena for nitrogen fixation under weak irradiance

1. In situ measurements of nitrogen fixation rates for Aphanizomenon in fertile Colorado lakes with low inorganic nitrogen concentrations demonstrated high efficiency of nitrogen fixation at low irradiance. 2. For study populations, rates of N₂ fixation in darkness and with alternating exposure to light and darkness were a higher percentage of light‐saturated rates for Aphanizomenon than for Anabaena, suggesting storage of reduced metabolites at high irradiance that are used subsequently by Aphanizomenon when cells are forced by mixing into zones of low irradiance. Also, saturation of N₂ fixation occurred over a lower range of irradiance for Aphanizomenon than for Anabaena. 3. High efficiency of N₂ fixation in Aphanizomenon at low or fluctuating irradiance is complementary to its previously demonstrated high efficiency of photosynthesis at low irradiance. Nitrogen fixation rate was also strongly related to DIN concentration; fixation was highest at low DIN (maximum < 5 μg L^?1) but was also most vulnerable to photoinhibition under such conditions. 4. The fixation capabilities of Aphanizomenon under weak or varying irradiance could explain its commonly observed domination over Anabaena when transparency is low and available nitrogen is scarce.     

Elevated levels of fibrinogen-derived endogenous citrullinated peptides in synovial fluid of rheumatoid arthritis patients

Introduction

Rheumatoid arthritis (RA) is an autoimmune disease characterized by inflammation of the joints and the presence of autoantibodies directed against proteins containing the non-standard arginine-derived amino acid citrulline. The protein fibrinogen, which has an essential role in blood clotting, is one of the most prominent citrullinated autoantigens in RA, particularly because it can be found in the inflamed tissue of affected joints. Here, we set out to analyze the presence of citrullinated endogenous peptides in the synovial fluid of RA and arthritic control patients.

Methods

Endogenous peptides were isolated from the synovial fluid of RA patients and controls by filtration and solid phase extraction. The peptides were identified and quantified using high-resolution liquid chromatography-mass spectrometry.

Results

Our data reveal that the synovial fluid of RA patients contains soluble endogenous peptides, derived from fibrinogen, containing significant amounts of citrulline residues and, in some cases, also phosphorylated serine. Several citrullinated peptides are found to be more abundantly present in the synovial fluid of RA patients compared to patients suffering from other inflammatory diseases affecting the joints.

Conclusions

The increased presence of citrullinated peptides in RA patients points toward a possible specific role of these peptides in the immune response at the basis of the recognition of citrullinated peptides and proteins by RA patient autoantibodies.     

Methyl Jasmonate Increases the Tropane Alkaloid Scopolamine and Reduces Natural Herbivory in Brugmansia suaveolens: Is Scopolamine Responsible for Plant Resistance?

The tropane alkaloid (TA) scopolamine is suggested to protect Brugmansia suaveolens (Solanaceae) against herbivorous insects. To test this prediction in a natural environment, scopolamine was induced by methyl jasmonate (MJ) in potted plants which were left 10?days in the field. MJ-treated plants increased their scopolamine concentration in leaves and herbivory decreased. These findings suggest a cause?Ceffect relationship. However, experiments in laboratory showed that scopolamine affect differently the performance of the specialist larvae of the ithomiine butterfly Placidina euryanassa (C. Felder & R. Felder) and the generalist fall armyworm Spodoptera frugiperda (J. E. Smith): the specialist that sequester this TA from B. suaveolens leaves was not negatively affected, but the generalist was. Therefore, scopolamine probably acts only against insects that are not adapted to TAs. Other compounds that are MJ elicited may also play a role in plant resistance against herbivory by generalist and specialist insects, and deserve future investigations.