Model-based analysis supports interglacial refugia over long-dispersal events in the diversification of two South American cactus species.

Pilosocereus machrisii and P. aurisetus are cactus species within the P. aurisetus complex, a group of eight cacti that are restricted to rocky habitats within the Neotropical savannas of eastern South America. Previous studies have suggested that diversification within this complex was driven by distributional fragmentation, isolation leading to allopatric differentiation, and secondary contact among divergent lineages. These events have been associated with Quaternary climatic cycles, leading to the hypothesis that the xerophytic vegetation patches which presently harbor these populations operate as refugia during the current interglacial. However, owing to limitations of the standard phylogeographic approaches used in these studies, this hypothesis was not explicitly tested. Here we use Approximate Bayesian Computation to refine the previous inferences and test the role of different events in the diversification of two species within P. aurisetus group. We used molecular data from chloroplast DNA and simple sequence repeats loci of P. machrisii and P. aurisetus, the two species with broadest distribution in the complex, in order to test if the diversification in each species was driven mostly by vicariance or by long-dispersal events. We found that both species were affected primarily by vicariance, with a refuge model as the most likely scenario for P. aurisetus and a soft vicariance scenario most probable for P. machrisii. These results emphasize the importance of distributional fragmentation in these species, and add support to the hypothesis of long-term isolation in interglacial refugia previously proposed for the P. aurisetus species complex diversification.

Polymorphic microsatellite markers for the rare and endangered cactus Uebelmannia pectinifera (Cactaceae) and its congeneric species.

The cactus genus Uebelmannia includes 3 narrow endemic species associated with rocky savanna habitats in eastern South America. Because of their rarity and illegal over-collection, all of these species are endangered. Taxonomic uncertainties resulting from dramatic local variation in morphology within Uebelmannia species preclude effective conservation efforts, such as the reintroduction or translocation of plants, to restore declining populations. In this study, we developed and characterized 18 perfect, dinucleotide simple-sequence repeat markers for U. pectinifera, the most widely distributed species in the genus, and tested the cross-amplification of these markers in the remaining congeneric species and subspecies. All markers were polymorphic in a sample from 2 U. pectinifera populations. The effective number of alleles ranged from 1.6 to 8.7, with an average per population of 3.3 (SE ± 0.30) and 4.5 (SE ± 0.50). Expected heterozygosity ranged from 0.375 to 0.847 and 8-10 loci showed departures from Hardy- Weinberg equilibrium in the analyzed populations. Based on the observed polymorphism level of each marker, as well as the analysis of null allele presence and evidence of amplification of duplicate loci, a subset of 12 loci can be used as reliable markers to investigate the genetic structure, diversity, and species limits of the Uebelmannia genus.

Usefulness of cpDNA markers for phylogenetic and phylogeographic analyses of closely related cactus species.

Although plastid DNA has been widely explored as a marker of choice for phylogeny and phylogeography studies, little is known about its utility for examining relationships between closely related species. The slow evolutionary rates inherent to chloroplast (cp) DNA make it difficult to perform lower level taxonomic analyses, particularly at the population level. We characterized the nucleotide variation and investigated the utility of eight noncoding cpDNA regions in four closely related species of the Pilosocereus aurisetus group (Cactaceae), an endemic taxon of eastern South America. The plastid intergenic spacers 5'-trnS-trnG, 3'-trnS-trnG and trnT-trnL were the most variable regions and were the most useful for lower level taxonomic comparisons, especially when used together. We conclude that an adequate combination of regions alongside indels as an additional character improves the usefulness of cpDNA for phylogenetic studies.

Pentachlorophenol (PCP) dechlorination in horizontal-flow anaerobic immobilized biomass (HAIB) reactors.

This study verifies the potential applicability of horizontal-flow anaerobic immobilized biomass (HAIB) reactors to pentachlorophenol (PCP) dechlorination. Two bench-scale HAIB reactors (R1 and R2) were filled with cubic polyurethane foam matrices containing immobilized anaerobic sludge. The reactors were then continuously fed with synthetic wastewater consisting of PCP, glucose, acetic acid, and formic acid as co-substrates for PCP anaerobic degradation. Before being immobilized in polyurethane foam matrices, the biomass was exposed to wastewater containing PCP in reactors fed at a semi-continuous rate of 2.0 microg PCP g(-1) VS. The applied PCP loading rate was increased from 0.05 to 2.59 mg PCP l(-1)day(-1) for R1, and from 0.06 to 4.15 mg PCP l(-1)day(-1) for R2. The organic loading rates (OLR) were 1.1 and 1.7 kg COD m(-3)day(-1) at hydraulic retention times (HRT) of 24h for R1 and 18 h for R2. Under such conditions, chemical oxygen demand (COD) removal efficiencies of up to 98% were achieved in the HAIB reactors. Both reactors exhibited the ability to remove 97% of the loaded PCP. Dichlorophenol (DCP) was the primary chlorophenol detected in the effluent. The adsorption of PCP and metabolites formed during PCP degradation in the packed bed was negligible for PCP removal efficiency.

Monitoring the trihalomethanes present in water after treatment with chlorine under laboratory condition.

In this work assays involving chlorinated water samples, which were previous spiked with humic substances or algae blue green and following the production of the THMs for 30 days is described. To implement the assays, five portions of 1,000 ml of water were stored in glass bottles. The water samples were treated with solutions containing 2, 3, 4 and 5 mg l(-1) chlorine. The samples aliquots (60 ml) were transferred into the glass vials, 10 ml were removed to have a headspace and 100 microl of the 10 mg l(-1) pentafluortoluene bromide solution was added to each vial. The extraction step was performed by adding 10 g of Na(2)SO(4) followed by 5 ml of n-pentane. The vials were stopped with a TFE-faced septum and sealed with aluminum caps. The generated THMs were determined by gas chromatography with electron capture detector using reference solutions with concentration ranging from 8 to 120 microg l(-1) THMs. Three assays were monitored during 30 days and chloroform was the predominant compound found in the water samples, while other species of THMs were not detected. The results showed that when the chlorine concentration was increased in water samples containing algae the concentration of THM varied randomly. Nevertheless, in water samples containing humic substances the increase of the THM concentration presented a relationship with the chlorine concentration. It was also observed that chloroform concentration increased with the elapsed time up to one and six days to water samples spiked with humic substances and algae blue green, respectively and decreased along 30 days. By other hand, assays performed using water samples containing decanted algae material showed that THM was not generated by the chlorine addition.

Preliminary notes on yeasts associated with necrotic cactus stems from different localities in Brazil

As leveduras encontradas em cladódios necrosados de três cactos colunares (Pilosocereus machrisii, Pilosocereus vilaboensis and Praecereus euchlorus) em oito localidades no Brasil foram identificadas e uma análise de similaridade baseada na distância Sorensen foi usada para comparar as comunidades nas diferentes localidades. Dos 56 cladódios necrosados amostrados, 32 produziram colônias de leveduras. Dez espécies de leveduras foram identificadas a partir de 53 isolados, com as espécies Pichia cactophila, Candida sonorensis, Geotrichum sp. e Sporopachydermia cereana sendo as mais comuns. As espécies restantes ocorreram em baixas freqüências. O dendograma (UPGMA) construído a partir da análise de similaridade agrupou comunidades de leveduras obtidas em diferentes espécies de cactos e indicou que as espécies de cactos hospedeiros não foram determinantes nesse agrupamento.

Wing shape heritability and morphological divergence of the sibling species Drosophila mercatorum and Drosophila paranaensis.

The fruit-flies Drosophila paranaensis and Drosophila mercatorum pararepleta are sibling species belonging to the repleta group. Females of these two species are normally considered to be morphologically indistinguishable while males only differ consistently in the morphology of their genitalia. These species are sympatric throughout a large area of their geographic distribution. In this study, we investigated the degree of morphological divergence between D. paranaensis and D. mercatorum pararepleta based on morphometric analysis of their wings. The ellipse method was used to describe the placement of the longitudinal and transversal wing veins as well as the size of the wing and the shape of its outline. The heritability under laboratory and field conditions was also estimated from the parameters generated. Multivariate analysis showed that wing morphology possessed sufficient differences to discriminate between the two species with a successful classification rate of 95-98% for females and 82-87% for males. The results of the autoclassification were confirmed by a cross-validation test for females (92-96%). Most measurements possessed significant natural heritability (a mean of 0.48 for D. mercatorum and 0.88 for D. paranaensis), indicating that the variation observed was related to differences in genes acting additively. The principal difference between the two species was in the placement of the posterior transverse wing vein. However, the pattern of morphological variation in the wings of both species was similar, possibly because of shared restrictions in wing development pathways.

Formaldehyde degradation in an anaerobic packed-bed bioreactor.

The development of appropriate technologies for the treatment of formaldehyde discharged into the environment is important to minimize its impact. Aerobic systems have been employed, although alternative anaerobic treatments have also been widely studied, mainly due to their low energy consumption and sludge production. However, toxic substances can lead to disturbances in anaerobic reactors. Some research has already been developed on formaldehyde anaerobic biological treatment, but no consensus has yet been reached about its behavior nor has the most efficient system been identified. Aiming at finding supporting evidence for this issue, therefore, this study investigated the degradation and toxicity of formaldehyde in a Horizontal-Flow Anaerobic Immobilized Sludge Reactor. Formaldehyde concentrations of 26.2-1158.6 mg HCHO/L were applied in the reactor, resulting in formaldehyde and chemical oxygen demand removal efficiencies of 99.7% and 92%, respectively. Volatile fatty acids with up to five carbons, found during the degradation of formaldehyde, are believed to indicate that the degradation followed routes unlike those suggested in the literature, which reports the formation of intermediates such as methanol and formic acid. The Monod kinetic model adhered to the experimental data well, with apparent kinetic parameters estimated as r(app)max) = 2.79 x 10(-3) mg HCHO/mg SSVh and K (app)(s) = 242.8 mg HCHO/L.

Antifungal susceptibility of clinical and environmental isolates of Cryptococcus neoformans to four antifungal drugs determined by two techniques.

A total of 64 Cryptococcus neoformans strains, including clinical and environmental Brazilian isolates var. neoformans and var. gattii, were tested for susceptibility to amphotericin B, 5-flucytosine, fluconazole and itraconazole. The tests were performed according to the recommendations of National Committee of Clinical Laboratory Standards and the method of macrodilution in liquid medium of Shadomy et al. [Manual de Microbiologia Clínica, 4th ed. Buenos Aires: Editorial Medica Panamericana, 1987: 1229-38]. For most drugs there was a significant difference between the readings taken at 24 and 48 h with both methods. When the minimum inhibitory concentrations obtained by the two techniques were compared, significant differences were observed for amphotericin B and fluconazole. Overall, differences in drug susceptibility with respect to the origin of the isolates or the variety of the fungus were not observed. As an exception, the gattii variety exhibited a high resistance rate to amphotericin B when the technique of Shadomy et al. was applied, a fact possibly related to the greater difficulty for treatment of the disease caused by this fungal variety.

Relationships between necrotic cactus availability and population size in a cactophilic Drosophila (Diptera, Drosophilidae) located on a sandstone table hill in Brazil

Drosophila gouveai is a cactophilic species endemic to South America. In southeast Brazil it is found on summits of isolated hills, which apparently are current refugia resulting from climatic changes during the Quaternary Period. It breeds only in necrotic cactus cladodes of Pilosocereus machrisii. Temporal differences in necrotic cactus availability could have a great impact upon D. gouveai population size, and could thus influence its evolutionary history. We analyzed the relationship between necrotic cactus availability and population size of D. gouveai. The fluctuation in the population size, variation in necrotic cactus availability and exploitation of this resource by larvae were surveyed bimonthly for one year on a sandstone table hill in central-south Brazil. Temporal necrotic cactus availability did not vary significantly, though in June there was a moderate decrease Larval populations were highest in October and December. The D. gouveai population size was highest in February and remained relatively stable the rest of the year. The observed fluctuation in population size was not a function of temporal necrotic cactus availability in quantitative terms.

Relationships between necrotic cactus availability and population size in a cactophilic Drosophila (Diptera, Drosophilidae) located on a sandstone table hill in Brazil.

Drosophila gouveai is a cactophilic species endemic to South America. In southeast Brazil it is found on summits of isolated hills, which apparently are current refugia resulting from climatic changes during the Quaternary Period. It breeds only in necrotic cactus cladodes of Pilosocereus machrisii. Temporal differences in necrotic cactus availability could have a great impact upon D. gouveai population size, and could thus influence its evolutionary history. We analyzed the relationship between necrotic cactus availability and population size of D. gouveai. The fluctuation in the population size, variation in necrotic cactus availability and exploitation of this resource by larvae were surveyed bimonthly for one year on a sandstone table hill in central-south Brazil. Temporal necrotic cactus availability did not vary significantly, though in June there was a moderate decrease Larval populations were highest in October and December. The D. gouveai population size was highest in February and remained relatively stable the rest of the year. The observed fluctuation in population size was not a function of temporal necrotic cactus availability in quantitative terms.

Hybrid reactor performance in pentachlorophenol (pcp) removal by anaerobic granules.

The present research aimed at evaluating pentachlorophenol (PCP) degradation in a hybrid reactor supplied with a mixture of fatty acids (propionic, butyric, acetic and lactic) and methanol. The performance of the reactor is remarkably stable and efficient during PCP additions at range of 2.0 to 21.0 mg/L. The reduction of chemical oxygen demand (COD) was around 97% and methane was found to be 88% in the biogas production. The efficiency of volatile fatty acids breakdown was 93%, 64% and 74% respectively for butyric, propionic and acetic. PCP total removal of more than 99% was reached by granular sludge activities formed during 21 months of reactor operation. Methanogenic microorganisms predominance was noticed with 10(5) to 10(6) cells/mL during enumeration on methanol or lactate added to sulfate culture media. The removal rate was 1.07 mg PCP.g-1 VS.d-1 during the highest PCP concentration addition.