Short-term storage in vitro and large-scale propagation of grapevine genotypes

The objective of this work was to evaluate the large-scale propagation of grapevine genotypes after short-term storage in vitro. Microshoots from ten grapevine genotypes were used. The following storage temperatures were evaluated: 10, 20, and 25°C. After short-term storage, the shoots were propagated in up to five successive subcultures, to assess the large-scale propagation of the germplasm maintained under conditions of minimal growth. The propagated shoots were rooted in different concentrations of indolbutiric acid (IBA) and acclimatized in greenhouse. The best temperature for short-term storage in vitro and survival of the genotypes was 20°C. In the propagation phase, the highest number of shoots per explant was found in the subcultures 4 and 5, with averages of 4.9 and 4.8 shoots per explant, respectively. In the rooting phase, the best results for number of roots were obtained using a culture medium supplemented with 0.4 µmol L-1 of IBA, with an average of three roots per shoot. During the acclimation phase, a survival rate higher than 95% was achieved after 30 days in the greenhouse. Grapevine genotypes maintained for six months in vitro, at 20ºC, can be micropropagated in large scale.

PRELIMINARY RESULTS OF A LARGE-SCALE TREE INVENTORY OF UPLAND RAIN FOREST IN THE CENTRAL AMAZON

A large-scale inventory of trees > 10cm DBH was conducted in the upland "terra firme" rain forest of the Distrito Agropecuário da SUFRAMA (Manaus Free Zone Authority Agricultural District) approximately 65Km north of the city of Manaus (AM), Srasil. Thegeneral appearance and structure of the forest is described together with local topography and soil texture. Thepreliminary results of the Inventory provide a minimum estimate of 698 tree species in 53 families in the 40Km radius sampled, including 17 undescribed species. Themost numerically abundant families, Lecythidaceae, Leguminosae, 5apotaceae and Burseraceae as also among the most species rich families. One aspect of this diverse assemblage is the proliferation of species within certain genera, Including 26 genera In 17 families with 6 or more species or morphospecies. Most species have very low abundances of less than 1 tree per hectare. While more abundant species do exist at densities ranging up to a mean of 12 trees per ha, many have clumped distributions leading to great variation in local species abundance. The degree of similarity between hectare samples based int the Coefficient of Community similarity Index varies widely over different sample hectares for five ecologically different families. Soil texture apparently plays a significant role In determining species composition in the different one hectare plots examined while results for other variable were less consistent. Greater differences in similarity indices are found for comparisons with a one hectare sample within the same formation approximately 40Km to the south. It is concluded that homogeneity of tree community composition within this single large and diverse yet continuous upland forest formation can not be assumed.

Large-scale production and purification of recombinant protein from an insect cell/baculovirus system in Erlenmeyer flasks: application to the chicken poly(ADP-ribose) polymerase catalytic domain

A simple and inexpensive shaker/Erlenmeyer flask system for large-scale cultivation of insect cells is described and compared to a commercial spinner system. On the basis of maximum cell density, average population doubling time and overproduction of recombinant protein, a better result was obtained with a simpler and less expensive bioreactor consisting of Erlenmeyer flasks and an ordinary shaker waterbath. Routinely, about 90 mg of pure poly(ADP-ribose) polymerase catalytic domain was obtained for a total of 3 x 109 infected cells in three liters of culture

The use of saliva as a practical and feasible alternative to urine in large-scale screening for congenital cytomegalovirus infection increasesinclusion and detection rates

INTRODUCTION: Although urine is considered the gold-standard material for the detection of congenital cytomegalovirus (CMV) infection, it can be difficult to obtain in newborns. The aim of this study was to compare the efficiency of detection of congenital CMV infection in saliva and urine samples. METHODS: One thousand newborns were included in the study. Congenital cytomegalovirus deoxyribonucleic acid (DNA) was detected by polymerase chain reaction (PCR). RESULTS: Saliva samples were obtained from all the newborns, whereas urine collection was successful in only 333 cases. There was no statistically significant difference between the use of saliva alone or saliva and urine collected simultaneously for the detection of CMV infection. CONCLUSIONS: Saliva samples can be used in large-scale neonatal screening for CMV infection.

Assessment of schistosomiasis in the semi-arid Northeast region of Brazil: the São Francisco River large-scale water transposition project

Abstract: INTRODUCTION Risk of schistosomiasis expansion to semi-arid northeastern Brazil under the influence of the Integration Project of the São Francisco River (IPSFR) was assessed. METHODS: Stool examinations of schoolchildren, epidemiological investigation, and survey of the local host snail Biomphalaria straminea were performed in five IPSFR municipalities. RESULTS Six of 4,770 examined schoolchildren were egg-positive for Schistosoma mansoni. Biomphalaria straminea was widespread, but not naturally infected with S. mansoni. Snails experimentally exposed to two laboratory S. mansoni strains yielded infection indices of 1-4.5%. CONCLUSIONS: There is evidence of active schistosomiasis transmission in the area; thus, intensive surveillance actions are required.

Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale

Malaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screening platform for pooled samples aimed at reducing both the time and the cost of these diagnoses. Positive and negative samples were analysed in individual and pooled experiments using real-time polymerase chain reaction (PCR), nested PCR and an immunochromatographic test. For the individual tests, 46/49 samples were positive by real-time PCR, 46/49 were positive by nested PCR and 32/46 were positive by immunochromatographic test. For the assays performed using pooled samples, 13/15 samples were positive by real-time PCR and nested PCR and 11/15 were positive by immunochromatographic test. These molecular methods demonstrated sensitivity and specificity for both the individual and pooled samples. Due to the advantages of the real-time PCR, such as the fast processing and the closed system, this method should be indicated as the first choice for use in large-scale diagnosis and the nested PCR should be used for species differentiation. However, additional field isolates should be tested to confirm the results achieved using cultured parasites and the serological test should only be adopted as a complementary method for malaria diagnosis.

Factor structure of Bech's version of the Brief Psychiatric Rating Scale in Brazilian patients

The objective of the present study was to evaluate the factor structure of Bech's version of the Brief Psychiatric Rating Scale (BPRS), translated into Portuguese. The BPRS was administered to a heterogeneous group of psychiatric inpatients (N = 98) and outpatients (N = 62) in a University Hospital. Each patient was evaluated from one to eight times. The interval between consecutive interviews was one week for the inpatients and one month for the outpatients. The results were submitted to factorial analysis. The internal consistency of the total scale and of each factor was also estimated. Factorial analysis followed by normalized orthogonal rotation (Varimax) yielded four factors: Withdrawal-Retardation, Thinking Disorder, Anxious-Depression and Activation. Internal consistency measured by Cronbach's alpha coefficient ranged from 0.766 to 0.879. The data show that the factor structure of the present instrument is similar to that of the American version of the BPRS which contains 18 items, except for the absence of the fifth factor of the latter scale, Hostile-Suspiciousness.

Cross-scale determinants of palm species distribution

We investigate palm species distribution, richness and abundance along the Mokoti, a seasonally-dry river of southeastern Amazon and compare it to the patterns observed at a large scale, comprising the entire Brazilian territory. A total of 694 palms belonging to 10 species were sampled at the Mokoti River basin. Although the species showed diverse distribution patterns, we found that local palm abundance, richness and tree basal area were significantly higher from the hills to the bottomlands of the study region, revealing a positive association of these measures with moisture. The analyses at the larger spatial scale also showed a strong influence of vapor pressure (a measure of moisture content of the air, in turn modulated by temperature) and seasonality in temperature: the richest regions were those where temperature and humidity were simultaneously high, and which also presented a lower degree of seasonality in temperature. These results indicate that the distribution of palms seems to be strongly associated with climatic variables, supporting the idea that, by 'putting all the eggs in one basket' (a consequence of survival depending on the preservation of a single irreplaceable bud), palms have become vulnerable to extreme environmental conditions. Hence, their distribution is concentrated in those tropical and sub-tropical regions with constant conditions of (mild to high) temperature and moisture all year round.

Diversity and structure of microcrustacean assemblages (Cladocera and Copepoda) and limnological variability in perennial and intermittent pools in a semi-arid region, Bahia, Brazil

Temporary wetlands undergo recurrent drought due to the scarcity of water, which disrupts the hydrological connectivity with adjacent aquatic systems. However, some environments retain water for longer periods, allowing greater persistence of the community. The current study evaluated differences in the microcrustacean assemblages and limnological variability between perennial and intermittent pools in a semi-arid region of Brazil. The abiotic features (water temperature, pH, total alkalinity, electrical conductivity and depth) of intermittent pools were affected more than perennial pools due to loss of water volume. This may have contributed to a higher average richness and diversity index in some intermittent pools and differences in the structure of the assemblages. The lowest species richness and diversity were recorded where physical factors, such as a large quantity of suspended solids and variability in the electrical conductivity of the water and pH, make the environment unsuitable for these organisms. These results suggest that community development in intermittent pools is interrupted by the dry season; when the water returns, due to rainfall or rising groundwater, each pond undergoes a different process of colonization. In these circumstances, the biological importance of temporary aquatic environments is clear, since such pools provide shelters and have an important role in the maintenance of the regional diversity of aquatic environments.

Structure of the salivary glands of the unfed female tick Amblyomma cajennense (Fabricius) (Acarina: Ixodidae)

Acini in in the salivary glands of female tick specimens of Amblyomma ajennense unfed at both postnymphal and adult phases, were studied. The salivary glands are consisted by three acini, one agranular and two granular. The agranular acini are directly attached to the anterior portion of the main salivary duct, consisting of cells without valve. A relatively large, clear, central cell occupies most of the alveolar midsection. The central cell is in contact with the acini lumen. Granular acini consist of approximately seven to fourteen cells (type II acini) or seven to sixteen (type III acini). The type II acini have three types of granular cells ("a", "b" and "c") and valve; the type III acini have another three types of granular cells ("d", "e" and "f") also presenting a valve.

Structure of the salivary glands of the unfed male tick Amblyomma cajennense (Fabricius) (Acarina: Ixodidae)

Acini in the salivary glands of unfed male Amblyomma cajennense of different ages, were studied. The salivary glands consist of one agranular and three granular acini types. The agranular acini are directly attached to the medial and anterior portion of the main salivary duct, and to some branches of the secondary ducts. A large, clear, central cell occupies the centre and this cell is in contact with the acinar lumen. There is no valve to the lumen. Granular acini consist of approximately six to fourteen cells (type II acini) or eight to thirteen (type III acini). The type II acini have three types of granular cells ("a", "b" and "c") and a valve: the type III acini have three types of granular cells ("d", "e" and "f" and a valve.

Derris (Lonchocarpus) urucu (Leguminosae) Extract Modifies the Peritrophic Matrix Structure of Aedes aegypti (Diptera:Culicidae)

Aqueous suspension of ethanol extracts of Derris (Lonchocarpus) urucu (Leguminosae), collected in the state of Amazonas, Brazil, were tested for larvicidal activity against the mosquito Aedes aegypti (Diptera:Culicidae). The aim of this study was to observe the alterations of peritrophic matrix in Ae. aegypti larvae treated with an aqueous suspension of D. urucu extract. Different concentrations of D. urucu root extract were tested against fourth instar larvae. One hundred percent mortality was observed at 150 µg/ml (LC50 17.6 µg/ml) 24 h following treatment. In response to D. urucu feeding, larvae excreted a large amount of amorphous feces, while control larvae did not produce feces during the assay period. Ultrastructural studies showed that larvae fed with 150 µg/ml of D. urucu extract for 4 h have an imperfect peritrophic matrix and extensive damage of the midgut epithelium. Data indicate a protective role for the peritrophic matrix. The structural modification of the peritrophic matrix is intrinsically associated with larval mortality.

Population genetic structure of the major malaria vector Anopheles darlingi (Diptera: Culicidae) from the Brazilian Amazon, using microsatellite markers

The population genetic structure of Anopheles darlingi, the major human malaria vector in the Neotropics, was examined using seven microsatellite loci from nine localities in central and western Amazonian Brazil. High levels of genetic variability were detected (5-25 alleles per locus; H E = 0.519-0.949). There was deviation from Hardy-Weinberg Equilibrium for 59.79% of the tests due to heterozygote deficits, while the analysis of linkage disequilibrium was significant for only two of 189 (1.05%) tests, most likely caused by null alleles. Genetic differentiation (F ST = 0.001-0.095; Nm = 4.7-363.8) indicates that gene flow is extensive among locations < 152 km apart (with two exceptions) and reduced, but not absent, at a larger geographic scale. Genetic and geographic distances were significantly correlated (R² = 0.893, P < 0.0002), supporting the isolation by distance (IBD) model. The overall estimate of Ne was 202.4 individuals under the linkage disequilibrium model, and 8 under the heterozygote excess model. Analysis of molecular variance showed that nearly all variation (~ 94%) was within sample locations. The UPGMA phenogram clustered the samples geographically, with one branch including 5/6 of the state of Amazonas localities and the other branch the Acre, Rondônia, and remaining Amazonas localities. Taken together, these data suggest little genetic structure for An. darlingi from central and western Amazonian Brazil. These findings also imply that the IBD model explains nearly all of the differentiation detected. In practical terms, populations of An. darlingi at distances < 152 km should respond similarly to vector control measures, because of high gene flow.

Fine structure of Henneguya hemiodopsis sp. n. (Myxozoa), a parasite of the gills of the Brazilian teleostean fish Hemiodopsis microlepes (Hemiodontidae)

A fish-infecting myxosporean, Henneguya hemiodopsis sp. n., found infecting the gills of Hemiodopsis microlepis and collected from the Poty River near the city of Teresina, Brazil, was described based on ultrastructural studies. The parasite occurred within large whitish polysporic plasmodia (up to 200 μm in diameter) containing asynchronous developmental sporogonic stages, mainly mature spores. The spores measured 19.7 ± 0.9 μm in total length (n = 30) and the ellipsoidal spore body was 10.8 ± 0.5 μm long, 3.3 ± 0.4 μm wide and 2.5 ± 0.5 μm thick. The spores were composed of two equal shell valves adhering together along the straight suture line, with each valve having equal-sized caudal tapering tails measuring 8.7 ± 0.6 μm in length. The spores were surrounded by a thin anastomosed network of microfibrils, more evident on the tails. There were two symmetric elongated bottle-like polar capsules 3.5 ± 0.3 μm long and 1.0 ± 0.2 μm wide, each with a polar filament with five to six coils. Given the morphological and ultrastructural differences from previously described parasites and the specificity of the host species, we propose a new species, named H. hemiodopsis sp. n.

Large-scale patterns in morphological diversity and species assemblages in Neotropical Triatominae (Heteroptera: Reduviidae)

We analysed the spatial variation in morphological diversity (MDiv) and species richness (SR) for 91 species of Neotropical Triatominae to determine the ecological relationships between SR and MDiv and to explore the roles that climate, productivity, environmental heterogeneity and the presence of biomes and rivers may play in the structuring of species assemblages. For each 110 km x 110 km-cell on a grid map of America, we determined the number of species (SR) and estimated the mean Gower index (MDiv) based on 12 morphological attributes. We performed bootstrapping analyses of species assemblages to identify whether those assemblages were more similar or dissimilar in their morphology than expected by chance. We applied a multi-model selection procedure and spatial explicit analyses to account for the association of diversity-environment relationships. MDiv and SR both showed a latitudinal gradient, although each peaked at different locations and were thus not strictly spatially congruent. SR decreased with temperature variability and MDiv increased with mean temperature, suggesting a predominant role for ambient energy in determining Triatominae diversity. Species that were more similar than expected by chance co-occurred near the limits of the Triatominae distribution in association with changes in environmental variables. Environmental filtering may underlie the structuring of species assemblages near their distributional limits.