Diversity of Paenibacillus durus strains isolated from soil and different plant rhizospheres evaluated by ARDRA and gyrB-RFLP analysis

Strains belonging to Paenibacillus durus isolated from the rhizosphere of various grasses and from bulk soil were previously divided into five phenotypic groups (A1–A5) based on the fermentation pattern of six carbohydrates (A1: sorbitol (+), A2: dulcitol and tagatose (+), A3: starch and glycogen (+), A4: starch, glycogen and d-arabitol (+) and A5: negative for these carbohydrates). This study aimed to assess whether plant types select for specific P. durus phenotypic groups. For that purpose, polymerase chain reaction-restriction fragment length polymorphism analysis of part of genes encoding 16S rRNA (ARDRA) and DNA gyrase subunit B (gyrB-RFLP) were used to produce genetic fingerprints. ARDRA and gyrB-RFLP data were clustered together to generate a dendrogram and two main clusters were observed. Cluster I showed a predominance of strains isolated from wheat, maize and sugarcane rhizospheres. Strains isolated from maize were distributed among the five patterns of carbohydrate metabolism, while strains isolated from sugarcane showed to be predominantly able to metabolize starch and glycogen. Neither sorbitol- nor arabitol-metabolizing strains were found in cluster II, which consisted of strains isolated from soil and from all plant species used. Our results suggest that the plants influenced the diversity of P. durus in their rhizospheres.     

Asymmetric segregation of polarized antigen on B cell division shapes presentation capacity

During the activation of humoral immune responses, B cells acquire antigen for subsequent presentation to cognate T cells. Here we show that after mouse B cells accumulate antigen, it is maintained in a polarized distribution for extended periods in vivo. Using high-throughput imaging flow cytometry, we observed that this polarization is preserved during B cell division, promoting asymmetric antigen segregation among progeny. Antigen inheritance correlates with the ability of progeny to activate T cells: Daughter cells receiving larger antigen stores exhibit a prolonged capacity to present antigen, which renders them more effective in competing for T cell help. The generation of progeny with differential capacities for antigen presentation may have implications for somatic hypermutation and class switching during affinity maturation and as B cells commit to effector cell fates.     

The molecular basis for recognition of bacterial ligands at equine TLR2, TLR1 and TLR6

TLR2 recognises bacterial lipopeptides and lipoteichoic acid, and forms heterodimers with TLR1 or TLR6. TLR2 is relatively well characterised in mice and humans, with published crystal structures of human TLR2/1/Pam3CSK4 and murine TLR2/6/Pam2CSK4. Equine TLR4 is activated by a different panel of ligands to human and murine TLR4, but less is known about species differences at TLR2. We therefore cloned equine TLR2, TLR1 and TLR6, which showed over 80% sequence identity with these receptors from other mammals, and performed a structure-function analysis. TLR2/1 and TLR2/6 from both horses and humans dose-dependently responded to lipoteichoic acid from Staphylococcus aureus, with no significant species difference in EC50 at either receptor pair. The EC50 of Pam2CSK4 was the same for equine and human TLR2/6, indicating amino acid differences between the two species’ TLRs do not significantly affect ligand recognition. Species differences were seen between the responses to Pam2CSK4 and Pam3CSK4 at TLR2/1. Human TLR2/1, as expected, responded to Pam3CSK4 with greater potency and efficacy than Pam2CSK4. At equine TLR2/1, however, Pam3CSK4 was less potent than Pam2CSK4, with both ligands having similar efficacies. Molecular modelling indicates that the majority of non-conserved ligand-interacting residues are at the periphery of the TLR2 binding pocket and in the ligand peptide-interacting regions, which may cause subtle effects on ligand positioning. These results suggest that there are potentially important species differences in recognition of lipopeptides by TLR2/1, which may affect how the horse deals with bacterial infections.     

Using computed tomography images to characterize the effects of soil compaction resulting from large machinery on three-dimensional pore characteristics in an opencast coal mine dump

Journal of Soils and Sediments - The use of heavy machinery in mining areas increases during mining and dumping of waste soils, causing severe soil compaction and potentially affecting soil...     

Black soybean promotes the formation of active components with antihepatoma activity in the fermentation product of Agaricus blazei

The antihepatoma activity and related active components in the fermentation products of Agaricus blazei (AB) cultured in the medium containing soybean (S) or black soybean (BS) were investigated. AB(BS)-pE and AB(S)-pE were the ethanolic extracts from the fermentation products of AB(BS) and AB(S), respectively. According to the IC 50 values, AB(BS)-pE (161.1 and 24.0 microg/mL for Hep 3B and Hep G2 cells, respectively) exhibited stronger cytotoxicities against hepatoma cells than AB(S)-pE (>200 and 99.9 microg/mL for Hep 3B and Hep G2 cells, respectively). AB(BS)-pE was separated by silica gel column chromatography and eluted with n-hexane/ethyl acetate/methanol gradient solvent system into 21 fractions. Fraction 3 [AB(BS)-pE-F3], eluted with n-hexane/ethyl acetate (97:3 and 19:1, v/v), was the most active fraction having inhibitory activity on the proliferation of Hep 3B and Hep G2 cells (IC 50 of 3.6 and 1.9 microg/mL, respectively). Three major compounds, compounds 1- 3, were further isolated from the AB(BS)-pE-F3 fraction by reversed-phase semipreparative high-performance liquid chromatography. Compounds 2 and 3 gave better antihepatoma activity than that of compound 1. The IC 50 values of compounds 2 and 3 were 2.8 and 4.5 microg/mL for Hep 3B cells and 1.4 and 2.0 microg/mL for Hep G2 cells, respectively. The structures of compounds 2 and 3 were identified by UV, IR, electron impact mass spectrometry, and (1)H and (13)C NMR to be blazeispirols A and C, respectively. Blazeispirols A and C existed in the mycelia but not in the broth and were more in AB(BS)-pE (49.9 +/- 8.9 and 14.2 +/- 2.4 mg/g, respectively) than AB(S)-pE (15.9 +/- 1.7 and 3.9 +/- 0.6 mg/g, respectively). Additionally, the result shows that the production of blazeispirols A and C was increased after cultivation in the medium containing black soybean on day 6 and reached the maximum on day 12, and the contents of blazeispirols A and C were negatively correlated with Hep 3B and Hep G2 cell viabilities ( r = -0.84 to -0.93, P < 0.01). It suggests that blazeispirols A and C could be used as biomarkers to produce the fermentation product of A. blazei with antihepatoma activity.     

Absence of three known benzimidazole resistance mutations in Trichostrongylus tenuis, a nematode parasite of avian hosts

Benzimidazole (BZ) resistance is widespread in nematode parasites of livestock, but very little is known about the levels of BZ resistance in parasites with avian hosts. We investigated BZ resistance in Trichostrongylus tenuis, a nematode parasite of red grouse, Lagopus lagopus scotica. BZ anthelmintics had been in use in this system for up to 15 years, yet existing phenotypic evidence for resistance was inconclusive. We screened 1530 individuals from 14 populations at the principal β-tubulin locus for BZ resistance (isotype 1, residue 200) and 940 of these at two further resistance sites (isotype 1, residue 167; isotype 2, residue 200). No BZ resistant genotypes were found. Alternative mechanisms may be responsible for BZ resistance in this system, or the method and timing of treatments may reduce selection pressure for BZ resistance by creating substantial refugia for susceptible genotypes.     

Comparison of protocols to synchronize estrus and ovulation in estrous-cycling and prepubertal beef heifers

The objective of the experiment was to compare follicular dynamics, ovulatory response to GnRH, and synchrony of estrus and ovulation among estrous-cycling and prepubertal beef heifers synchronized with a controlled internal drug-release (CIDR)- based or GnRH-PGF(2alpha) (PG) protocol. Estrous-cycling beef heifers were randomly assigned to 1 of 4 treatments (C1, C2, C3, C4), and prepubertal beef heifers were randomly assigned to 1 of 2 treatments (P1, P2) by age and BW. Blood samples were taken 10 and 1 d before treatment to confirm estrous cyclicity status (progesterone > or =0.5 ng/mL estrous cycling). The CIDR Select (C1, n = 12; P1, n = 14)-treated heifers received a CIDR insert (1.38 g of progesterone) from d 0 to 14, GnRH (100 microg, i.m.) on d 23, and PG (25 mg, i.m.) on d 30. Select Synch + CIDR (C2, n = 12; P2, n = 11)-treated heifers received a CIDR insert and GnRH on d 23 and PG at CIDR removal on d 30. The CIDR-PG (C3, n = 12)-treated heifers received a CIDR insert on d 23 and PG at CIDR removal on d 30. Select Synch (C4, n = 12)-treated heifers received GnRH on d 23 and PG on d 30. HeatWatch transmitters were fitted at CIDR removal (C1, C2, C3, P1, and P2) or at GnRH administration (C4) for estrus detection. Ultrasound was used to determine the response to GnRH and the timing of ovulation after estrus. Among the estrous-cycling heifers, ovulatory response to GnRH and estrous response did not differ (P > 0.05). Among the prepubertal heifers, more (P = 0.02) P1 heifers responded to GnRH than P2 heifers, but estrous response did not differ (P > 0.05). Among the estrous-cycling heifers, variance for interval to estrus after PG was reduced (P < 0.05) for C1 compared with each of the other treatments, and C3 [corrected] was reduced (P < 0.05) compared with C2 [corrected] Variance for interval to ovulation after PG was reduced (P < 0.05) for C1 compared with each of the other treatments. Among the prepubertal heifers, there was no difference (P > 0.05) in variance for interval to estrus or ovulation. Results from C1 and P1 (T1) and C2 and P2 (T2) were combined to compare T1 and T2 among mixed groups of estrous-cycling and prepubertal heifers. Response to GnRH was greater (P < 0.01; 81% T1 and 39% T2), and variances for interval to estrus and ovulation for T1 were reduced (P < 0.01) compared with T2. In summary, CIDR Select improved (P < 0.01) the synchrony of estrus and ovulation compared with Select Synch + CIDR.     

Gonadotropin-releasing hormone-induced ovulation and luteinizing hormone release in beef heifers: effect of day of the cycle

The COSynch protocol has been used to synchronize ovulation and facilitate fixed-time AI in beef cattle. Establishment and maintenance of pregnancy was negatively affected, in previous studies, by GnRH-induced ovulation of small dominant follicles (/=10 mm) and increased ovulatory response after GnRH 2.     

Hepatic intranuclear glycogen inclusions in western barred bandicoots (Perameles bougainville).

The western barred bandicoot, Perameles bougainville, is an endangered Australian marsupial species. Routine histology of liver samples collected at necropsy from 19 of 20 (95%) western barred bandicoots revealed the sporadic to common occurrence of abnormal hepatocyte nuclei characterized by margination of chromatin and concomitant central pallor. Some abnormal hepatocyte nuclei were mildly to markedly enlarged and irregularly shaped. Periodic acid-Schiff reagent stained 131 of 142 (92%) of these abnormal hepatocyte nuclei. Positive staining was completely eliminated by diastase pretreatment. Transmission electron microscopy revealed that abnormal hepatocyte nuclei with marginated chromatin did not contain viral particles. Rather, glycogen beta-particles and alpha-rosettes were identified within some abnormal hepatocyte nuclei. Glycogen intranuclear inclusions were an incidental finding in western barred bandicoot hepatocytes.