Phthalates production from Curvularia senegalensis (Speg.) Subram, a fungal species associated to crops of commercial value

The fungal species Curvularia senegalensis was isolated from a soil sample collected at a Brazilian region of cerrado transition. This microorganism was grown in vitro and the extract of the culture medium was fractionated by chromatographic methods yielding an oil rich in phthalates, from which seven derivatives were identified by infrared, (1)H and (13)C NMR and mass spectrometry as 1-hexyl-2-propylphthalate, 1-ethyl-2-heptylphthalate, 1-hexyl-2-butylphthalate, 1-heptyl-2-proylphthalate, 1-propyl-2-nonylphthalate and two positional isomers of 1-decyl-2-butane phthalate. This is the first report on the phthalates production by Curvularia senegalensis revealing a scientific basis for the use of this species on biodegradation experiments. Since C. senegalensis is a very common pathogen in some commercial crops, presence of highly toxic phthalates on the final feed products should be investigated.     

Different Effects of Transgenic Maize and Nontransgenic Maize on Nitrogen-Transforming Archaea and Bacteria in Tropical Soils

The composition of the rhizosphere microbiome is a result of interactions between plant roots, soil, and environmental conditions. The impact of genetic variation in plant species on the composition of the root-associated microbiota remains poorly understood. This study assessed the abundances and structures of nitrogen-transforming (ammonia-oxidizing) archaea and bacteria as well as nitrogen-fixing bacteria driven by genetic modification of their maize host plants. The data show that significant changes in the abundances (revealed by quantitative PCR) of ammonia-oxidizing bacterial and archaeal communities occurred as a result of the maize host being genetically modified. In contrast, the structures of the total communities (determined by PCR-denaturing gradient gel electrophoresis) were mainly driven by factors such as soil type and season and not by plant genotype. Thus, the abundances of ammonia-oxidizing bacterial and archaeal communities but not structures of those communities were revealed to be responsive to changes in maize genotype, allowing the suggestion that community abundances should be explored as candidate bioindicators for monitoring the possible impacts of cultivation of genetically modified plants.     

Detection of a Bacterium Associated with a Leaf Spot Disease of Maize in Brazil

A leaf spot disease of maize occurring in Brazil in the 1980s was described as being caused by the ascomycete Phaeosphaeria maydis (P. Henn) Rane. Payak and Renfro (imperfect form Phyllosticta sp.). Disease symptoms were dark-green water-soaked spots that later became necrotic lesions. There are no reports at present in the literature of re-infection by the fungus under controlled conditions, casting doubt on the true identity of the pathogen. In this study, cytological analyses of lesions at the initial stages did not detect the presence of fungal structures. Bacterial colonies with yellow pigmentation were isolated from the lesions, which reacted positively in hypersensitivity tests in tobacco plants. Maize plants were inoculated with the isolated bacteria. After 72 h incubation in a dew chamber, plants were transferred to a greenhouse, where they remained until evaluation. Typical symptoms of the disease were observed 5–7 days after inoculation of plants, only on treatments inoculated with the bacteria. The bacterium was re-isolated, which suggests its involvement in the initial phases of disease. The bacterium was identified as Pantoea ananas (synonym Erwinia ananas ).     

A new species of Backusella (Mucorales) from a Cerrado reserve in Southeast Brazil

Cerrado (Brazilian savanna) areas present a high number of endemic and unknown species and consequently have been included among the world’s biodiversity hotspots. After a survey of zygomycetes from Cerrado areas of the “Reserva Biológica de Mogi Guaçu” (RBMG) in São Paulo State, we have found a Mucor-like fungus that produces very large sporangiospores and clusters of giant cells visible to the naked-eye. This isolate was characterized morphologically, based on partial LSU (28S) and complete ITS rDNA sequences. Maximum likelihood phylogenetic analyses support its inclusion within the recently emended genus Backusella.     

Sensor and biosensor based on Prussian Blue modified gold and platinum screen printed electrodes

Gold (Au) and platinum (Pt) screen-printed electrodes were modified with Prussian Blue (PB) for the development of amperometric sensors selective for hydrogen peroxide detection. The sensors exhibited sensitivities towards H(2)O(2) equal to 2 A M(-1) cm(-2) for Au and 1 A M(-1) cm(-2) for Pt electrodes. The sensors were also employed as the basis for construction of glucose biosensors through further modification with crystallised glucose oxidase immobilised in a Nafion membrane. In order to improve the operational stability of the modified electrodes a buffer solution containing tetrabutylammonium toluene-4-sulfonate was used. The long-term performance of the sensors and biosensors were evaluated by continuous monitoring of hydrogen peroxide and glucose solutions (50 microM and 1 mM, respectively) in the flow-injection mode for 10 h.     

Enzyme replacement therapy for Mucopolysaccharidosis Type I among patients followed within the MPS Brazil Network

Mucopolysaccharidosis type I (MPS I) is a rare lysosomal disorder caused by deficiency of alpha-L-iduronidase. Few clinical trials have assessed the effect of enzyme replacement therapy (ERT) for this condition. We conducted an exploratory, open-label, non-randomized, multicenter cohort study of patients with MPS I. Data were collected from questionnaires completed by attending physicians at the time of diagnosis (T1; n = 34) and at a median time of 2.5 years later (T2; n = 24/34). The 24 patients for whom data were available at T2 were allocated into groups: A, no ERT (9 patients; median age at T1 = 36 months; 6 with severe phenotype); B, on ERT (15 patients; median age at T1 = 33 months; 4 with severe phenotype). For all variables in which there was no between-group difference at baseline, a delta of ≥ ± 20% was considered clinically relevant. The following clinically relevant differences were identified in group B in T2: lower rates of mortality and reported hospitalization for respiratory infection; lower frequency of hepatosplenomegaly; increased reported rates of obstructive sleep apnea syndrome and hearing loss; and stabilization of gibbus deformity. These changes could be due to the effect of ERT or of other therapies which have also been found more frequently in group B. Our findings suggest MPS I patients on ERT also receive a better overall care. ERT may have a positive effect on respiratory morbidity and overall mortality in patients with MPS I. Additional studies focusing on these outcomes and on other therapies should be performed.     

Phylogenetic and physiological characterization of organic waste-degrading bacterial communities

One of the greatest challenges in contemporary society is to reduce and treat household solid waste. The choice of inoculum to be used for start-up in reactors that degrade organic waste is critical to the success of organic waste treatment. In this study, the functional diversity, phylogenetic identification, and biogas production of bacterial communities from six inoculum sources were investigated. We used BIOLOG EcoPlates to evaluate the metabolic abilities of the bacterial communities, followed 16S rRNA gene sequence analysis to determine the phylogenetic affiliation of the bacteria responsible for carbon consumption. We observed great diversity in the physiological profiles. Of the six inocula tested, the sludge from an upflow anaerobic sludge blanket reactor (SRU) contained the most diverse, metabolically versatile microbiota and was characterized by the highest level of biogas production. By contrast, the sludge of the anaerobic lagoon (SAL) showed the worst performance in BIOLOG EcoPlates assays, but it exhibited the most diversity and generated the second largest amount of biogas. The bacterial isolates retrieved from BIOLOG EcoPlates were characterized as aerobic and/or facultative anaerobic, and were mainly Gram-negative. Phylogenetic analysis revealed that the isolates belonged to three major phyla: Proteobacteria, Firmicutes and Actinobacteria, represented by 33 genera. Proteobacteria exhibited the most diversity. The distribution of the bacterial genera differed considerably among the six inocula. Pseudomonas and Bacillus, which are able to degrade a wide range of proteins and carbohydrates, predominated in five of the six inocula. Analysis of the bacterial communities in this study indicates that both SRU and SAL microbiota are candidates for start-up inocula in anaerobic reactors. These start-up inocula must be studied further in order to identify their practical applications in degrading organic waste.     

Diversity of Paenibacillus spp. in the rhizosphere of four sorghum (Sorghum bicolor) cultivars sown with two contrasting levels of nitrogen fertilizer assessed by rpoB-based PCR-DGGE and sequencing analysis

The diversity of Paenibacillus species was assessed in the rhizospheres of four cultivars of sorghum sown in Cerrado soil amended with two levels of nitrogen fertilizer (12 and 120 kg/ha). Two cultivars (IS 5322-C and IS 6320) demanded the higher amount of nitrogen to grow, whereas the other two (FBS 8701-9 and IPA 1011) did not. Using the DNA extracted from the rhizospheres, a Paenibacillus-specific PCR system based on the RNA polymerase gene (rpoB) was chosen for the molecular analyses. The resulting PCR products were separated into community fingerprints by DGGE and the results showed a clear distinction between cultivars. In addition, clone libraries were generated from the rpoB fragments of two cultivars (IPA 1011 and IS 5322-C) using both fertilization conditions, and 318 selected clones were sequenced. Analyzed sequences were grouped into 14 Paenibacillus species. A greater diversity of Paenibacillus species was observed in cultivar IPA 1011 compared with cultivar IS 5322-C. Moreover, statistical analyses of the sequences showed that the bacterial diversity was more influenced by cultivar type than nitrogen fertilization, corroborating the DGGE results. Thus, the sorghum cultivar type was the overriding determinative factor that influenced the community structures of the Paenibacillus communities in the habitats investigated.     

Temporal dynamics of microbial communities in the rhizosphere of two genetically modified (GM) maize hybrids in tropical agrosystems

The use of genetically modified (GM) plants still raises concerns about their environmental impact. The present study aimed to evaluate the possible effects of GM maize, in comparison to the parental line, on the structure and abundance of microbial communities in the rhizosphere. Moreover, the effect of soil type was addressed. For this purpose, the bacterial and fungal communities associated with the rhizosphere of GM plants were compared by culture-independent methodologies to the near-isogenic parental line. Two different soils and three stages of plant development in two different periods of the year were included. As evidenced by principal components analysis (PCA) of the PCR-DGGE profiles of evaluated community, clear differences occurred in these rhizosphere communities between soils and the periods of the year that maize was cultivated. However, there were no discernible effects of the GM lines as compared to the parental line. For all microbial communities evaluated, soil type and the period of the year that the maize was cultivated were the main factors that influenced their structures. No differences were observed in the abundances of total bacteria between the rhizospheres of GM and parental plant lines.     

Synthesis and biological evaluation of thiazolidinedione derivatives with high ligand efficiency to P. aeruginosa PhzS

The thiazolidinone ring is found in compounds that have widespan biology activity and there is mechanism-based evidence that compounds bearing this moiety inhibit P. aeruginosa PhzS (PaPzhS), a key enzyme in the biosynthesis of the virulence factor named pyocyanin. Ten novel thiazolidinone derivatives were synthesised and screened against PaPhzS, using two orthogonal assays. The biological results provided by these and 28 other compounds, whose synthesis had been described, suggest that the dihydroquinazoline ring, found in the previous hit (A- Kd = 18 µM and LE = 0.20), is not required for PaPzhS inhibition, but unsubstituted nitrogen at the thiazolidinone ring is. The molecular simplification approach, pursued in this work, afforded an optimised lead compound (13- 5-(2,4-dimethoxyphenyl)thiazolidine-2,4-dione) with 10-fold improvement in affinity (Kd= 1.68 µM) and more than 100% increase in LE (0.45), which follows the same inhibition mode as the original hit compound (competitive to NADH).

Executive summary

• PhzS is a key enzyme in the pyocyanin biosynthesis pathway in P. aeruginosa.
• Orthogonal assays (TSA and FITC) show that fragment-like thiazolidinedione derivatives bind to PaPhzS with one-digit micromolar affinity.
• Fragment-like thiazolidinedione derivatives bind to the cofactor (NADH) binding site in PaPhzS.
• The molecular simplification optimised the ligand efficiency and affinity of the lead compound.