Phenotypic characterization and in vitro antifungal sensitivity of Candida spp. and Malassezia pachydermatis strains from dogs

Yeasts of the genera Candida and Malassezia can be found as commensal microorganisms in animals. The main species of importance in veterinary medicine are Malassezia pachydermatis and Candida albicans. The objectives of this study were to conduct a phenotypic characterization and to evaluate the in vitro antifungal sensitivity of strains of C. albicans (n=5), C. tropicalis (n=3) and M. pachydermatis (n=32) isolated from dogs. The phenotyping was based on macro and micromorphological features as well as biochemical analysis. The techniques of microdilution in broth and dilution in agar were used to evaluate the in vitro sensitivity of Candida spp. and M. pachydermatis, respectively. The tested drugs were ketoconazole (KTC), itraconazole (ITC), fluconazole (FLC) and amphotericin B (AMB). The morphological analysis of the strains of Candida spp. and M. pachydermatis did not show any noteworthy alterations when compared to standard strains. On the other hand, in the biochemical tests, 34.4% of the strains of M. pachydermatis were negative for the urease test. Four strains of C. albicans were resistant to FLC with a minimum inhibitory concentration (MIC) >64microg/mL and all were resistant to KTC and ITC (MIC>16microg/mL). The MIC for two strains of C. tropicalis were >16microg/mL for KTC and ITC, and >64microg/mL for FLC. It is worth highlighting that all of the strains tested were sensitive to AMB with the MIC varying from 0.25-1.0microg/mL. All strains of M. pachydermatis were sensitive to ITC with a minimum fungistatic concentration (MFC) 0.0075microg/mL. The MIC for 29 strains was the same (MFC0.0075microg/mL) for KTC. The MFCs for FLC varied from 1 to 16microg/mL, and for AMB, the MFC interval was 0.125-8microg/mL. There were no alterations in the classic phenotypic features of the strains of Candida spp. and M. pachydermatis isolated from dogs but, unlike M. pachydermatis, Candida spp. were much more resistant to azole antifungal agents.     

Allocation patterns and phenology in wild and selected accessions of annual and perennial Physaria (Lesquerella, Brassicaceae)

Two processes that determine a good performance of plants in arid environments are phenology and resource allocation patterns. With a longer growing season and larger allocation to root, perennials achieve better access to resources and are more resilient to stresses than annuals. In traditional agricultural systems selection for optimal soil nutrient uptake has been a secondary breeding objective, because crops receive subsidies of water and fertilizers. However in arid lands, caution is required during domestication, to avoid changes in structural traits which may be the basis for sustainable production. Due to inherent differences in hierarchy among annual and perennial species, we propose that the changes in phenology and allocation brought about by selection will depend on the life cycle. We performed field studies comparing wild and selected accessions of annual and perennial species of Physaria. Life cycle determined the functional basis of seed yield. In annuals, selection resulted in early anthesis (1 week earlier), a lower allocation to roots and leaves (twofold lower), and an increase in harvest index (an increase of 62%). Selected perennials had higher biomass at maturity (45% higher), linked to a longer reproductive period (3 weeks longer) than their wild relatives. The vegetative allocation found in wild perennials remained unchanged after selection. While annuals selected for seed yield could compromise the capacity for acquisition of resources, selection in perennials did not modify the allocation strategy responsible for their positive adjustment to low resource environments. We found a trade off between seed yield potential and yield stability that resulted in lower performance of selected accessions in low quality environments in relation to their wild relatives.     

Problems related to seed production in the African region

Improved seeds can make a substantial contribution to agricultural productivity in African countries. Seeds are one of the most important sources of innovation, particularly in resource-constrained small farm environments. They carry the genetic potential of the crops, determining the upper limit on yield and, therefore, the ultimate productivity of other inputs. Furthermore, new roles for seeds are rapidly recognized all over the world for the delivery systems of many innovative biotechnological products and as carriers of plant protection chemicals, biologicals and growth regulators. The seed sector development in Africa varies considerably among countries. Rather effective and diversified seed industries have slowly emerged and are operational in a few countries (e.g. Egypt, Kenya, Zimbabwe). In a growing number of other countries (e.g. Malawi, Zambia) the seed production and supply system has developed reasonably well in some areas for some crops. However, in most countries (e.g. Cameroon, Ethiopia, Ghana, Tanzania and Uganda) progress has been very limited in spite of investments and assistance. During the last decade a number of countries have undergone privatisation and/or seed market liberalisation as potential solutions to under-performance in the seed sector. However, it is not certain that this will produce a practical improvement and solve the problems of the poor rural majority of the population. In this paper the main problems related to the seed sector in Africa are discussed together with the possible roles of the private and public sectors in developing and maintaining an efficient and active seed industry. This revised version was published online in July 2006 with corrections to the Cover Date.     

Isolation and partial characterization of a pectic polysaccharide from the fruit pulp of Spondias cytherea and its effect on peritoneal macrophage activation

The total carbohydrate content of the intact pulp of Spondias cytherea was 41%. Polysaccharides were obtained via hot aqueous extraction after defatting with organic solvents. The aqueous extract was treated with excess ethanol to form a precipitate, which was then solubilized in water. The material precipitated upon acidification when HCl was removed. The resulting supernatant fraction was submitted to freeze-thawing treatment yielding a soluble fraction (sFTS). This fraction had Ara, Rha, Gal and GalA in its structure as determined by GC-MS. 13C NMR analysis showed signals assigned to alpha-L-Araf, beta-D-Galp, alpha-D-GalpA and alpha-L-Rhap units, in addition to galacturonic acid units, which were present also as methyl ester. These results suggest a type I rhamnogalacturonan with arabinogalactan branches. Cell eliciting activity in a dose-depending pattern was observed in vitro on peritoneal macrophages treated with sFTS.     

A New Application for <Emphasis Type="Italic">Klebsiella oxytoca</Emphasis> in Bioremediation: Treatment of Manganese-Laden Wastewaters

Bioremediation is a method of removing manganese from wastewaters, and many bacterial species with a role in manganese bioremediation have been identified over several decades. Using K medium with Mn(II), a bacterial consortium was obtained from water sample collected from a mine drainage in southeastern Brazil (Minas Gerais, Brazil), and the isolates were identified as Klebsiella oxytoca using both biochemical and 16S rRNA phylogenetic analysis. Among such isolates, sample SA8 was tested for its manganese removal capability during a week-long experiment in K medium amended with manganese sulfate. Growth of isolate SA8 produced an increase in pH concomitantly to Mn(II) removal, which was not observed in the abiotic control. Manganese removal occurred through oxidation as confirmed by reaction with leucoberbelin blue, and therefore, a new potential application for K. oxytoca in mine water bioremediation is proposed.     

Essential oils from Mediterranean lamiaceae as weed germination inhibitors

The essential oils obtained from rosemary (Rosmarinus officinalis L.), thyme (Thymus vulgaris L.), and savory (Satureja montana L.) and the four monoterpenes that are their major constituents have been analyzed by GC and GC-MS and tested for their allelopathic properties on the seeds of three different annual weeds (Chenopodium album, Portulaca oleracea, and Echinochloa crus-galli) and three crops (Raphanus sativus, Capsicum annuum, and Lactuca sativa), with the aim to evaluate in vitro their potential as germination inhibitors. The essential oil composition varied with the species, thymol being the main constituent (44%) of thyme and carvacrol (57%) that of savory oil. Differences in essential oil composition were observed within two different rosemary ecotypes, type A, with alpha-pinene (37%) and 1,8-cineole (23%), and type B, characterized by a 2-fold content of 1,8-cineole (47%). This latest essential oil inhibited completely the germination of weeds while concurrently displaying little effect on pepper. The other two oils showed less selective action. S. montana essential oil, with 57% carvacrol, is the most active compound, completely inhibiting germination both of crops and weeds. Borneol, one of the main constituents of the oil of rosemary type B, showed an activity comparable to that of the whole oil. Crop and weed seeds treated with 1,8-cineole showed germination values that were not significantly different from controls, even if a slowing of the germination process expressed in terms of a significant increase in mean germination time was observed. Monoterpene compounds also present in the essential oils mainly represented the volatile fraction released from the crops and their residues into the soil.     

Usefulness of volunteer data to measure the large scale decline of “common” toad populations

Measuring a species decline is pivotal to evaluate their conservation status, but an accurate assessment of demographic trends requires observations collected across broad spatial and temporal scales. Volunteers can help to collect information over large scales, but their data may be affected by heterogeneity for sampling efforts and protocols, which may influence detection probability. Ignoring this issue may conduct to misleading conclusions. Here we show that data collected by different volunteer groups can be integrated with measures of sampling efforts, to obtain information on large scale demographic trends. We collected data on 33 common toad (Bufo bufo) populations across Italy for the period 1993–2010. We used two approaches (meta-analysis; analysis of average change in population size) to evaluate the overall demographic trend. We incorporated measures of volunteer sampling efforts into analyses, to take into account changes in detection probability. Toad abundance significantly declined in the last decade. From 2000 to 2010, 70% of populations showed a strong decline, and only 10% increased. Trends were heterogeneous among populations, but taking into account sampling effort reduced heterogeneity by 40%. We detected a 76% cumulative average decline of toad populations, despite an increasing mean sampling effort. The widespread toad decline rises concern for its future, also because the causes remain unclear. Volunteer data can be extremely useful to identify large scale population trends, if information on sampling effort are recorded and used to adjust counts.     

Structure of genetic diversity among common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) varieties of mesoamerican and andean origins using new developed microsatellite markers

A common bean genomic library was constructed using the ‘IAC-UNA’ variety enriched for (CT) and (GT) for microsatellite motifs. From 1,209 sequenced clones, 714 showed microsatellites distributed over 471 simple and 243 compound motifs. GA/CT and GT/CA were the most frequent motifs found among these sequences. A total of 123 microsatellites has been characterized. Out of these, 87 were polymorphic (73.7%), 33 monomorphic (26.8%), and 3 (2.4%) did not amplify at all. In a sample of 20 common bean materials selected from the Agronomic Institute Germplasm Bank, the number of alleles per locus varied 2–9, with an average of 2.82. The polymorphic information content (PIC) of each marker varied from 0.05 to 0.83, with a 0.45 average value. Cluster and principal coordinate analysis of the microsatellite data were consistent with the original assignment of the germplasm accessions into the Andean and Mesoamerican gene pools of common bean. Low polymorphism levels detected could be associated with the domestication process. These microsatellites could be a valuable resource for the bean community because of their use as new markers for genetic studies. Electronic supplementary material The online version of this article (doi: ) contains supplementary material, which is available to authorized users. An erratum to this article can be found at