Diversity of Chromobacterium violaceum isolates from aquatic environments of state of Pará, Brazilian Amazon

The present study intended to characterize the phenotypic and genetic diversity of Brazilian isolates of Chromobacterium violaceum from aquatic environments within the Amazon region. Nineteen isolates showed morphological properties of C. violaceum and the majority grew at 44°C. Low temperatures, in contrast, showed to be inhibitory to their growth, as eleven isolates did not grow at 10ºC and nine did not produce pigmentation, clearly indicating an inhibition of their metabolism. The largest variation among isolates was observed in the citrate test (Simmons), in which 12 isolates were positive, and in the oxidation/fermentation of sucrose, with six positives isolates. Chloramphenicol, gentamicin and sulfonamides efficiently inhibited bacterial growth. Amplified products of the recA gene were digested with HindII or PstI, which produced three or four restriction fragments patterns, respectively. The combined analysis arranged the isolates into six genospecies. The higher diversity observed in Belém (genotypes C, D, E and F) may be a consequence of intense human occupation, pollution of the aquatic environment or due to the higher diversity of the environments sampled in that region. In conclusion, a high level of genetic and phenotypic diversity was observed, and four new genospecies were described.

Stool sample storage conditions for the preservation of Giardia intestinalis DNA

Stool is chemically complex and the extraction of DNA from stool samples is extremely difficult. Haemoglobin breakdown products, such as bilirubin, bile acids and mineral ions, that are present in the stool samples, can inhibit DNA amplification and cause molecular assays to produce false-negative results. Therefore, stool storage conditions are highly important for the diagnosis of intestinal parasites and other microorganisms through molecular approaches. In the current study, stool samples that were positive for Giardia intestinalis were collected from five different patients. Each sample was stored using one out of six different storage conditions [room temperature (RT), +4ºC, -20ºC, 70% alcohol, 10% formaldehyde or 2.5% potassium dichromate] for DNA extraction procedures at one, two, three and four weeks. A modified QIAamp Stool Mini Kit procedure was used to isolate the DNA from stored samples. After DNA isolation, polymerase chain reaction (PCR) amplification was performed using primers that target the β-giardin gene. A G. intestinalis-specific 384 bp band was obtained from all of the cyst-containing stool samples that were stored at RT, +4ºC and -20ºC and in 70% alcohol and 2.5% potassium dichromate; however, this band was not produced by samples that had been stored in 10% formaldehyde. Moreover, for the stool samples containing trophozoites, the same G. intestinalis-specific band was only obtained from the samples that were stored in 2.5% potassium dichromate for up to one month. As a result, it appears evident that the most suitable storage condition for stool samples to permit the isolation of G. intestinalis DNA is in 2.5% potassium dichromate; under these conditions, stool samples may be stored for one month.

Cover plants and mineral nitrogen: effects on organic matter fractions in an oxisol under no-tillage in the cerrado

Cover plants are essential for the sustainability of no-tillage systems in tropical regions. However, information on the effects of these plants and N fertilization on soil organic matter fractions is still scarce. This study evaluated the effect of cover crops with different chemical composition and of N topdressing on the labile and humified organic matter fractions of an Oxisol of the Cerrado (savanna-like vegetation). The study in a randomized complete block design was arranged in split-plots with three replications. Four cover species were tested in the plots and the presence or absence of N topdressing in the subplot. The following cover species were planted in succession to corn for eight years: Urochloa ruziziensis; Canavalia brasiliensis M. ex Benth; Cajanus cajan (L.) Millsp; and Sorghum bicolor (L.) Moench. In general, the cultivation of U. ruziziensis increased soil C levels, particularly of C in the humic acid and particulate organic C fractions, which are quality indicators of soil organic matter. The C in humic substances and mineral organic C accounted for the highest proportions of total organic C, demonstrating the strong interaction between organic matter, Fe and Al oxides and kaolinite, which are predominant in these weathered soils of the Cerrado.

Follicular dynamics in zebu cattle

The aim of this study was to investigate the follicular dynamics during estrous cycle in Gir breed (Bos indicus) cows. Follicular growth and atresia during estrous cycle were evaluated using a portable ultrasound device. Luteal activity was evaluated by serum progesterone levels. Cycles with two (6.67%), three (60.00%), four (26.67%) and five (6.67%) follicular waves were observed. There was no difference (P>0.05) in dominant or subordinate follicles growth or atresia rates among follicular waves. The maximum diameter of the ovulatory follicle was higher than the diameter of the other dominant follicles in cycles with four waves, and higher than the diameter of the second dominant follicle in cycles with three waves (P<0.05). There was no difference (P>0.05) in estrous cycle length (21.11±1.76 and 22.25±1.71 days) or progesterone levels during diestrous (4.48±1.45 and 5.08±1.40 ng/mL) between cycles with three or four waves. Follicular dynamics in Gir cattle is characterized by a higher incidence of cycles with three or four waves, associated with a low persistence of the dominant follicle.

Allohexaploid pearl millet x elephantgrass population potential for a recurrent selection program

The objective of this work was to evaluate the potential of allohexaploid pearl millet x elephantgrass (HGL) population for a recurrent selection program through open-pollinated progenies. Seventy-eight progenies, one representative sample of the population, and two commercial cultivars, Pioneiro and Paraíso, were evaluated in a 9x9 triple lattice design, in two sites. Plant height and dry matter yield were evaluated in three and four cuts, respectively. For plant height, the 17 best progenies were similar to both commercial controls, while for dry matter yield they were higher than 'Paraíso' and lower than 'Pioneiro'. The correlation between progenies and cuts indicated that the fourth cut represents the mean of all cuts, and the possibility of using early selection. Heritability estimates considering cuts and sites were 56.9% for plant height and 58.8% for dry matter yield, and the expected response to selection was 23.4% for dry matter yield and 18.1% for plant height. These results demonstrate the promising HGL population potential for a recurrent selection program.

Modeling growth and yield of loblolly pinestands under intensive management

Abstract:The objective of this work was to develop and validate a prognosis system for volume yield and basal area of intensively managed loblolly pine (Pinus taeda) stands, using stand and diameter class models compatible in basal area estimates. The data used in the study were obtained from plantations located in northern Uruguay. For model validation without data loss, a three-phase validation scheme was applied: first, the equations were fitted without the validation database; then, model validation was carried out; and, finally, the database was regrouped to recalibrate the parameter values. After the validation and final parameterization of the models, a simulation of the first commercial thinning was carried out. The developed prognosis system was precise and accurate in estimating basal area production per hectare or per diameter classes. There was compatibility in basal area estimates between diameter class and whole stand models, with a mean difference of -0.01 m2ha-1. The validation scheme applied is logic and consistent, since information on the accuracy and precision of the models is obtained without the loss of any information in the estimation of the models' parameters.

Quince 'CPP': new dwarf rootstock for pear trees on organic and high density planting

In Brazil, pear production presents the same incipient situation over the last 15 years, due mostly to low production technology. In this context, this study aimed to evaluate the development, growth and production of the pear tree cultivars Cascatense, Tenra and Hosui grafted on 'CPP' quince rootstock, using 'FT' pear as interstem. This trial was carried out in Guarapuava, State of Paraná, Southern region of Brazil, by five productive cycles. The pear trees were planted in September of 2004, spaced at 1.0 x 4.0 m (2,500 trees ha-1), trained to the modified central leader, on a Four-wire trellis, with drip irrigation and cultivated under organic production system. The following variables were evaluated: sprouting, anthesis, yield, fruit weight, soluble solids content, titratable acidity, pulp firmness, canopy area per plant and per hectare and trunk diameter. The pear tree cv. Tenra was outstanding most of the years for fruit yield, and, consequently, showed the highest accumulated yield over the period (51.6 t ha-1), followed by the cultivars Cascatense (39.7 t ha-1) and Hosui (18.7 t ha-1). All pear cultivars presented suitable physical-chemical characteristics for commercial purposes, with minimal average soluble solids content of 11% at harvest. The maximum canopy area per hectare was attained for cv. Cascatense (3063.2 m²), that was considered insufficient for a high yield. These results suggest the needs for studies with higher density planting and other training systems, searching optimize canopy volume. One of the most limiting factors in the organic pear orchard was the incidence of pear dieback caused by Botriosphaeria dothidea, severe more often in pear trees cv. Hosui.

Voltametria de micropartículas imobilizadas: fundamentos e aplicações analíticas

The theoretical aspects of Voltammetry of Immobilized Microparticles (VIM) were discussed. The immobilization of microparticles on electrode surface, the electrode cleaning processes and the electrode materials were analyzed. The three-phase electrode model and the possible reactions between the immobilized particle and the electrolytic solution were discussed. In addition, this work discusses some selected applications of VIM published in the last years.

DFT study on low molecular weight α,α-ditert-butyl-4H-cyclopenta[2,1-b,3;4-b']dithiophene and α,α-ditert-butyl-4H-cyclopenta[2,1-b,3;4-b']dithiophene S-oxide bridged derivatives

The equilibrium geometries of α,α-ditert-butyl-4H-cyclopenta[2,1-b,3;4-b']dithiophene (DBDT) and α,α-ditert-butyl-4H-cyclopenta[2,1-b,3;4-b']dithiophene S-oxide (DBDTO) were studied at the DFT level of theory with a standard 6-311G* basis set. The molecular structures of the DBDT series were more planar than the corresponding DBDTO series, as revealed by dihedral angles. The UV-visible absorption calculated at TD-DFT/6-311G* showed two absorption peaks for all the molecules except C=S and C=O bridged molecules. In DBDTOs, C=S and C=O bridged molecules showed three and four absorption peaks, respectively. The DBDTOs had lower band gaps and longer wavelengths compared to the corresponding DBDTs.

Quantitative analysis of two important epidemiological features of the common bean: Phaeoisariopsis griseola pathosystem

This work quantifies two important epidemiological features of the bean (Phaseolus vulgaris)/Phaeoisariopsis griseola pathosystem. The first is the effect of the number of nights of leaf wetness on infection efficiency. Infection efficiency was below 10% when inoculated leaflets were exposed to less than two nights of leaf wetness. Optimum infection efficiencies were obtained after three to four nights of leaf wetness, at about 50%. Further nights of leaf wetness did not increase the infection efficiency. The second feature quantified is the relative rate of leaflet defoliation for varying levels of angular leaf spot severity. It increased with disease severity according to a logarithm-like curve, and a relative rate of 0.23 day-1 was estimated for a severity of 18%. The implications of these results on the disease epidemiology are discussed.

Pharmacological immunomodulation enhances peripheral nerve regeneration

To assess the effect of N-Acetylmuramyl-L-Alanyl-D-Isoglutamine MDP topically administrated on the regenerating peripheral neurons, twelve male C57BL/6J adult mice were equally distributed into three groups. Four mice underwent unilateral sciatic nerve transection and polyethylene tubulization, with a 4mm gap between the proximal and distal nerve stumps and were implanted with collagen + PBS (COL). Other four animals underwent the same surgical procedure but received collagen + MDP (COL/MDP) inside the prosthesis. Four animals were not operated and served as control group (NOR). After 4 weeks, the regenerated nerve cables were processed for total myelinated axon counting and myelinated fiber diameter measurement. The L5 dorsal root ganglion (DRG) was also removed and sectioned for sensory neurons counting and measurement. The results revealed significant difference (p<0.05) in axonal counting among the groups NOR (4,355±32), COL (1,869±289) and COL/MDP (2,430±223). There was a significant reduction in the axonal diameter in the operated groups (COL=3.38µm±1.16 and COL/MDP=3.54µm±1.16) compared to NOR (6.19µm±2.45). No difference was found in the number of DRG neurons between the experimental groups (COL=564±51; COL/MDP=514±56), which presented fewer sensory neurons compared to NOR (1,097±142). Data obtained indicate that locally applied MDP stimulates peripheral nerve regeneration in mice.

Molecular Basis for Resistance to Fluazifop-P-Butyl in Itchgrass (Rottboellia cochinchinensis) from Costa Rica

Rottboellia cochinchinensis is an annual grass weed species known as itchgrass, or "caminadora" in America´s Spanish speaking countries, and has become a major and troublesome weed in several crops. The application of fluazifop-P-butyl at recommended rates (125 g a.i. ha-1) was observed to be failing to control itchgrass in a field in San José, Upala county, Alajuela province, Costa Rica. Plants from the putative resistant R. cochinchinensis population survived fluazifop-P-butyl when treated with 250 g a.i. ha-1 (2X label rate) at the three- to four-leaf stage under greenhouse conditions. PCR amplification and sequencing of partial carboxyl transferase domain (CT) of the acetyl-CoA carboxylase (ACCase) gene were used to determine the molecular mechanism of resistance. A single non-synonymous point mutation from TGG (susceptible plants) to TGC (putative resistant plants) that leads to a Trp-2027-Cys substitution was found. This Trp-2027-Cys mutation is known to confer resistance to all aryloxyphenoxyproprionate (APP) herbicides to which fluazifop-P-butyl belongs. To the best of our knowledge, this is the first report of fluazifop-P-butyl resistance and a mutation at position 2027 for a Costa Rican R. cochinchinensis population.

The antioxidant action of Polypodium leucotomos extract and kojic acid: reactions with reactive oxygen species

Two natural products Polypodium leucotomos extract (PL) and kojic acid (KA) were tested for their ability to scavenge reactive oxygen species (·OH, ·O2-, H2O2, ¹O2) in phosphate buffer. Hydroxyl radicals were generated by the Fenton reaction, and the rate constants of scavenging were 1.6 x 10(9) M-1 s-1 for KA and 1.0 x 10(9) M-1 s-1 for PL, similar to that of ethanol (1.4 x 10(9) M-1 s-1). With superoxide anions generated by the xanthine/hypoxanthine system, KA and PL (0.2-1.0 mg/ml) inhibited ·O2-dependent reduction of nitroblue tetrazolium by up to 30 and 31%, respectively. In the detection of ¹O2 by rose bengal irradiation, PL at 1.0 mg/ml quenched singlet oxygen by 43% relative to azide and KA by 36%. The present study demonstrates that PL showed an antioxidant effect, scavenging three of four reactive oxygen species tested here. Unlike KA, PL did not significantly scavenge hydrogen peroxide.

Sialic acid changes in Dalton's lymphoma-bearing mice after cyclophosphamide and cisplatin treatment

Sialic acid changes in Dalton's lymphoma cells and other tissues of 10-12-week-old Swiss albino mice were investigated in relation to tumour growth in vivo and following cyclophosphamide (ip, 200 mg/kg body weight) or cisplatin (ip, 8 mg/kg body weight) treatment. Three to four animals of both sexes were used in each experimental group. The sialic acid level of tumour cells (0.88 µmol/g) increased with tumour progression (1.44-1.59 µmol/g; P<=0.05) in mice. Sialic acid concentration in other tissues (liver, kidney, testes and brain) also increased (~40, 10, 30 and 58%, respectively) in the tumour-bearing hosts as compared with that in the respective tissues of normal mice. In vivo cyclophosphamide or cisplatin treatment resulted in an overall decrease of sialic acid contents in the tissues. Cyclophosphamide was more efficient in lowering tissue sialic acid than cisplatin (P<=0.01, ANOVA). It is suggested that sialic acid residues could be an important factor contributing to the manifestation of malignant properties in cancer cells in general and Dalton's lymphoma cells in particular. A significant decrease in the sialic acid content of Dalton's lymphoma cells after cisplatin or cyclophosphamide treatment may bring about specific changes in tumour cells which could be associated with tumour regression.

The effect of feeding on the respiratory activity of the sloth

The aim of the present study was to confirm whether feeding influences the resting breathing rate and to observe possible alterations in blood gas and pH levels produced by feeding in unanesthetized sloths (Bradypus variegatus). Five adult male sloths (4.1 ± 0.6 kg) were placed daily in an experimental chair for a period of at least 4 h for sitting adaptation. Five measurements were made for each sloth. However, the sloths one, two and five were studied once and the sloths three and four were studied twice. Breathing rate was determined with an impedance meter and the output signal was digitized. Arterial blood samples were collected for blood gas analysis with a BGE electrolytes analyzer and adjusted for the animal's body temperature and hemoglobin content. The data are reported as mean ± SD and were collected during the resting period (8:00-10:00 h) and during the feeding period (16:00-18:00 h). The mean breathing rate increased during mastication of ymbahuba leaves (rest: 5.0 ± 1, feeding: 10 ± 1 bpm). No significant alterations were observed in arterial pH (rest: 7.42 ± 0.05, feeding: 7.45 ± 0.03), PCO2 (rest: 35.2 ± 5.3, feeding: 33.3 ± 4.4 mmHg) or PO2 (rest: 77.5 ± 8.2, feeding: 78.4 ± 5.2 mmHg) levels. These results indicate that in unanesthetized sloths 1) feeding evokes an increase in breathing rate without a significant change in arterial pH, PCO2 or PO2 levels, and 2) the increase in breathing rate produced by feeding probably is due to the act of mastication.