Sporulation

Bacillus subtilis spo0K mutants are blocked at the first step in sporulation. The spo0K strain was found to contain two mutations: one was linked to the trpS locus, and the other was elsewhere on the chromosome. The mutation linked to trpS was responsible for the sporulation defect (spo-). The unlinked mutation enhanced this sporulation deficiency but had no phenotype on its own. The spo- mutation was located in an operon of five genes highly homologous to the oligopeptide transport (Opp) system of Gram-negative species. Studies with toxic peptide analogues showed that this operon does indeed encode a peptide-transport system. However, unlike the Opp system of Salmonella typhimurium, one of the two ATP-binding proteins, OppF, was not required for peptide transport or for sporulation. The OppA peptide-binding protein, which is periplasmically located in Gram-negative species, has a signal sequence characteristic of lipo-proteins with an amino-terminal lipo-amino acid anchor. Cellular location studies revealed that OppA was associated with the cell during exponential growth, but was released into the medium in stationary phase. A major role of the Opp system in Gram-negative bacteria is the recycling of cell-wall peptides as they are released from the growing peptidoglycan. We postulate that the accumulation of such peptides may play a signaling role in the initiation of sporulation, and that the sporulation defect in opp mutants results from an inability to transport these peptides.

Biomonitoring programs to access the ecological integrity of freshwaters tend to rely exclusively on structural parameters. Here we evaluated stream ecological integrity using (a) benthic macroinvertebrate derived metrics and a biotic index as measures of structural integrity and (b) oak litter decomposition and associated fungal sporulation rates as measures of functional integrity. The study was done at four sites (S1, S2, S3 and S4) along a downstream increasing phosphorus and habitat degradation gradient in a small stream. The biotic index, invertebrate metrics, invertebrate and fungal communities' structure and sporulation rates discriminated upstream and downstream sites. Decomposition rates classified sites S4 and S2 as having a compromised ecosystem functioning. Although both functional and structural approaches gave the same results for the most impacted site (S4), they were complementary for moderately impacted sites (S2 and S3), and we therefore support the need for incorporating functional measures in evaluations of stream ecological integrity.

Media for yeast identification tests were incorporated into the wells of a microtitre tray. The tests included fermentation and assimilation of carbohydrates, assimilation of nitrogen compounds, growth in vitamin-free medium, resistance to cycloheximide, and observations for cell morphology and sporulation. Results of tests conducted in the trays showed very good agreement with those obtained by conventional methods. Eighteen reference yeasts were correctly identified from tests conducted in the trays. The trays of media could be stored, and provided a convenient system for yeast identification.

Ulocladium atrum (strain 385) consistently reduced Botrytis cinerea sporulation on necrotic fragments of strawberry leaves. On these tissues, two strains of U. atrum (isolates 18558 and 18559) showed lower antagonistic activities than the reference strain 385. Colonization of strawberry leaflets by the three U. atrum strains appeared similar in the absence of B. cinerea, whether quantified by chitin or immunological assays. The second method (based on anti-U. atrum antibodies) revealed that strawberry leaflet colonization by U. atrum 385 was better than by the other U. atrum strains in the presence of B. cinerea. An immunoassay using anti-B. cinerea antibodies revealed that the colonization of B. cinerea in tissues was lower in the presence of U. atrum 385 than with the two other U. atrum strains. The enzymatic activities produced by U. atrum 385 during the colonization phases of necrotic tissues were compared to B. cinerea and U. atrum strains 18558 and 18559. U. atrum 385 had the ...

Coprological examinations of eight Ruppell’s agamas Agama rueppelli (Vaillant) revealed the presence of a coccidium of the genus Isospora Schneider, 1881 that represents a previously undescribed species. Oöcysts of Isospora farahi n. sp. are spherical or subspherical, 29.1 (26–31) × 28.8 (26–31) μm, with a shape-index of 1.01 (1–1.07). An oöcyst residuum, polar granules and micropyle are absent. The oöcyst wall is bilayered, brownish and smooth, c. 1.5–2 μm thick. The sporocysts are oval, 16.6 (15–18) × 11.4 (11–12) μm, with a shape-index of 1.46 (1.25–1.64) and both Stieda and substieda bodies. A sporocyst residuum is present as medium-sized granules scattered irregularly among the sporozoites. The sporozoites are vermiform, with a large posterior spherical refractile body. Endogenous development is intranuclear in the epithelial cells of the small intestine. Sporulation is unknown, as oöcysts were recovered from the faeces.

Visceral leishmaniasis is a zoonosis whose primary vector in Brazil is the sandfly Lutzomyia longipalpis Lutz & Neiva. Presently, efforts to control the vector have not been effective in reducing the prevalence of disease. A possible alternative to current strategies is the biological control of the vector using entomopathogenic fungi. This study evaluates the effects of the fungus, Beauveria bassiana (Bals.) Vuilleman, in different developmental stages of L. longipalpis. Five concentrations of the fungus were utilized ranging from 104 to 108 conidia/ml, with appropriate controls. The unhatched eggs, larvae and dead adults exposed to B. bassiana were sown to reisolate the fungus. The fungus was subsequently identified by polymerase chain reaction (PCR) and DNA sequencing. Exposure to B. bassiana reduced the number of eggs that hatched by 59% (P < 0.01). The longevity of infected adults was 5 days, significantly lower than that of the negative control which was 7 days (P < 0.001). The longevity of the adult sandfly exposed to the positive chemical (pyrethroid, cypermetherin) control was less than 1 day. The effects of fungal infection on the hatching of eggs laid by infected females were also significant and dose-dependent (P < 0.05). With respect to fungal post-infection growth parameters, only germination and sporulation were significantly higher than the fungi before infection (P < 0.001). The identity of the reisolated fungus was confirmed by automated DNA sequencing post-passage in all insect stages. These data show that B. bassiana has good pathogenic potential, primarily on L. longipalpis larvae and adults. Consequently, the use of this fungus in sandfly control programs has potential in reducing the use of chemical insecticides, resulting in benefits to humans and the environment.

Histidine kinases are sophisticated molecular sensors that are used by bacteria to detect and respond to a multitude of environ- mental signals. KinA is the major histidine kinase required for initiation of sporulation upon nutrient deprivation in Bacillus subtilis. KinA has a large N-terminal region (residues 1 to 382) that is uniquely composed of three tandem Per-ARNT-Sim (PAS) domains that have been proposed to constitute a sensor module. To further enhance our understanding of this “sensor” region, we defined the boundaries that give rise to the minimal autonomously folded PAS domains and analyzed their homo- and het- eroassociation properties using analytical ultracentrifugation, nuclear magnetic resonance (NMR) spectroscopy, and multiangle laser light scattering. We show that PASA self-associates very weakly, while PASC is primarily a monomer. In contrast, PASB forms a stable dimer (Kd [dissociation constant] of <10 nM), and it appears to be the main N-terminal determinant of KinA dimerization. Analysis of KinA mutants deficient for one or more PAS domains revealed a critical role for PASB, but not PASA, in autophosphorylation of KinA. Our findings suggest that dimerization of PASB is important for keeping the catalytic domain of KinA in a functional conformation. We use this information to propose a model for the structure of the N-terminal sensor mod- ule of KinA.