Escherichia coli heat-stable enterotoxin b (STb) in vivo internalization within rat intestinal epithelial cells

Heat-stable enterotoxin b (STb) is a low molecular weight toxin known to bind sulfatide, its receptor. The fate of STb bound to rat intestinal epithelium cells was followed using an anti-toxin gold labeled assay and transmission electron microscopy. The data suggest that STb toxin and the fusion protein maltose binding protein (MBP)-STb were internalized whereas its mutant I41 E-M42R with reduced hydrophobicity did not show internalization. There was a significant difference in the mean of gold particles per field between rat intestine incubated with STb or the fusion protein MBP-STb and the negative control consisting of intestine incubated with PBS alone. No subcellular compartment seems to be particularly aimed by the toxin as gold particles were randomly distributed within the cell.     

Isolation and characterization of Babesia pecorum sp. nov. from farmed red deer (Cervus elaphus)

The diversity of Babesia species infecting cervids in parts of central and southern Spain was analyzed by collecting blood from farmed red deer (Cervus elaphus). Babesia sp. was isolated in vitro from two red deer herds in Cádiz and Ciudad Real. The number of Babesia sp. carriers differed between the two herds: 36/77 in Cádiz and 1/35 in Ciudad Real. Hyalomma lusitanicum was the most prevalent tick species identified on the Cádiz farm vegetation and on sampled animals, and is therefore a candidate vector. The molecular characteristics of 21 isolates were determined by complete (8 isolates) or partial (13 isolates) 18S rRNA gene sequencing. The sequences were highly similar (over 99.4% identity) and 6 sequence types were identified at the level of one herd only, demonstrating a rather high genetic diversity. They formed a monophyletic clade, and members of the three main sequence types shared a similar morphology and the same erythrocyte susceptibility pattern. This clade also included Babesia sp. Xinjiang isolated from sheep in China and Babesia sp. identified in giraffe in South Africa, with identities higher than 98.3% and statistically relevant phylogenetic support. None of the biological properties analyzed for both Babesia from red deer and Babesia sp. Xinjiang allowed their differentiation (ability to develop in vitro in erythrocytes from cattle and sheep, as well as in erythrocytes from different cervids, unsuccessful infection of calves). We propose the Babesia isolated from red deer as a new species named B. pecorum. Whether Babesia sp. Xinjiang and the Babesia characterized in South Africa belong to the same species is debated.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0078-7) contains supplementary material, which is available to authorized users.     

Genome-wide insertional mutagenesis of Arabidopsis thaliana

Over 225,000 independent Agrobacterium transferred DNA (T-DNA) insertion events in the genome of the reference plant Arabidopsis thaliana have been created that represent near saturation of the gene space. The precise locations were determined for more than 88,000 T-DNA insertions, which resulted in the identification of mutations in more than 21,700 of the approximately 29,454 predicted Arabidopsis genes. Genome-wide analysis of the distribution of integration events revealed the existence of a large integration site bias at both the chromosome and gene levels. Insertion mutations were identified in genes that are regulated in response to the plant hormone ethylene.     

Impact of Sewage Sludge Compost Utilization on Chemical Properties of Olive Grove Soils

Field experiments were conducted for four years, between 1998 and 2002, in two olive grove soils of adult olive orchards (Olea europaea L. cv. Cornicabra) in a clay loam soil in Seseña (Toledo, Spain) and in a sandy loam soil in Aranjuez (Madrid, Spain). There were four treatments, sewage sludge compost (SSC), sewage sludge compost plus urea (SSC+U), urea (U) and control (C). Each treatment was replicated four times and two depths were studied (0-15 and 15-30 cm). Once a year, before spreading sewage sludge compost, soil samples were taken at depths of 0 to 15 cm and 15 to 30 cm. Organic matter, total Kjeldhal nitrogen, phosphorus availability, pH, and electric conductivity were measured. No differences were found between treatment on organic matter and electric conductivity after four years of application of sewage sludge compost to two olive grove soils. In relation to nitrogen content, sewage sludge compost, only in Seseña, produced higher nitrogen soil content than the traditional urea treatment. Sewage sludge compost applied on olive grove soils improved the Phosphorus availability for the olive tree. In Aranjuez, the use of sewage sludge compost increased the pH of the soil with respect to Urea and Control plots. In Seseña, the reverse effect was found.     

Fork reversal and ssDNA accumulation at stalled replication forks owing to checkpoint defects

Checkpoint-mediated control of replicating chromosomes is essential for preventing cancer. In yeast, Rad53 kinase protects stalled replication forks from pathological rearrangements. To characterize the mechanisms controlling fork integrity, we analyzed replication intermediates formed in response to replication blocks using electron microscopy. At the forks, wild-type cells accumulate short single-stranded regions, which likely causes checkpoint activation, whereas rad53 mutants exhibit extensive single-stranded gaps and hemi-replicated intermediates, consistent with a lagging-strand synthesis defect. Further, rad53 cells accumulate Holliday junctions through fork reversal. We speculate that, in checkpoint mutants, abnormal replication intermediates begin to form because of uncoordinated replication and are further processed by unscheduled recombination pathways, causing genome instability.     

The effect of Saccharomyces cerevisiae on digestion and mortality in the volcano rabbit (Romerolagus diazi)

An experiment was conducted to evaluate whether supplementation with a probiotic could enhance digestion and reduce mortality in the volcano rabbit in captivity. Two enclosures at Chapultepec Zoo, Mexico(114 individuals) were used in a crossover design(two periods of 60 days) with the following treatments: control group and supplementation with Saccharomyces cerevisiae(2×108 CFU/exhibit/day). Supplementation with the probiotic negatively affected(P<0.01) the digestibility of dry matter, organic matter, neutral detergent fiber(NDF) and energy. Mortality increased(P<0.04) following supplementation with the probiotic(4.26% vs. 8.89%), primarily in the juvenile rabbits. The results indicate that yeast supplementation in the volcano rabbit negatively affects digestion and mortality in captivity.     

Detection of Rhodococcus equi by polymerase chain reaction using species-specific nonproprietary primers.

Species-specific primers for the polymerase chain reaction (PCR) for the detection of Rhodococcus equi were developed. These primers were based on unique DNA fragments produced from R. equi reference strains and field isolates. Following random amplification of polymorphic DNA from R. equi and R. rhodochrous with a set of 40 arbitrary 10-base pair (bp) primers, a pair of species-specific primers was designed to detect a unique 700-bp fragment of R. equi chromosomal DNA. This PCR product was limited to R. equi and was not detectable in other Rhodococcus species or in a panel of additional gram-positive and gram-negative bacteria.     

Molecular diversity of Brazilian strains of porcine circovirus type 2 (PCV-2)

Porcine circovirus 2 (PCV-2) is associated with a broad range of syndromes. In this study, eight pig tissue samples from two Brazilian states were analyzed using six PCR primer pairs amplifying a 1705-bp fragment of the PCV-2 genome. The NJ distance-based method was used for the phylogenetic analysis with the eight field strains herein, 15 GenBank sequences and using PCV-1 as an out-group. This yielded two major clusters (A and B) for this viral species, with the Brazilian strains segregating with European and Asian sequences. Nucleotide identity was 99.7 to 100% among the sequences. This information can be used in further studies of pathogenesis related to PCV-2 in Brazil.     

Apparent electrical conductivity in dry versus wet soil conditions in a shallow soil

The general objective of this study was to evaluate the stability of patterns of apparent soil electrical conductivity (EC_a) in dry versus wet soil conditions in a shallow soil typically used for pastures in Mediterranean conditions of the southern region of Portugal. A 6 ha experimental field of permanent bio-diverse pasture was divided into 76 squares of 28 × 28 m. The soil electrical conductivity was measured using a Dualem 1S sensor under dry conditions (June 2007) and under wet conditions during the rainy season (March 2010). Soil samples, geo-referenced with GPS, were collected in a depth range of 0–0.30 m. The soil was characterized in terms of bedrock depth, moisture content, texture, pH, organic matter content, and macronutrients (nitrogen, phosphorus, and potassium). Pasture samples, also geo-referenced with GPS, were collected to measure the pasture dry matter yield. The statistical analysis of apparent electrical conductivity between dry and wet soil conditions resulted in a linear significant correlation coefficient (R = 0.88). The results also showed a significant correlation between apparent electrical conductivity and the relative field elevation (R = ?0.64 and R = ?0.66), the pasture dry matter yield (R = 0.42 and R = 0.48), the bedrock depth (R = 0.40 and R = 0.27), the pH (R = 0.50 and R = 0.49), the silt (R = 0.27 and R = 0.38) and soil moisture content (R = 0.48 and R = 0.45), in dry and wet conditions, respectively. A multi-variate regression was carried out using the following soil parameters that showed significant correlation with EC_a and that did not present multi-collinearity: pH, bedrock depth, silt and moisture content. The results showed, in dry and wet conditions, that the analysis was significant (R = 0.75 and R = 0.84, respectively). Overall, these results indicate the temporal stability of EC_a patterns under different soil moisture contents, which is relevant with respect to the time when a field should be surveyed and is important for using the electrical conductivity sensor, as a decision support tool for management zones in precision agriculture.