MICROORGANISMS IN ORGANIC AND NON ORGANIC HONEY SAMPLES OF AFRICANIZED HONEYBEES

This research was carried out to evaluate and compare 11 organic honey samples and six non organic honey samples, respectively, harvested from islands of the triple frontier (Sao Paulo, Parana and Mato Grosso do Sul states) and from the state of Parana, Brazil. The samples were studied for the presence of coliforms from 35 degrees C, to 45 degrees C and the enumeration of moulds and yeast, a minimum of 1.9 x 10(2) and a maximum of 1.1 x 10(3) CFU/g were observed in organic honey and a minimum of 1.8 x 10(1) and a maximum of 2.5 x 10(2) CFU/g were in non organic honey. In this studied region, the organic honey presented a microbiological quality inferior to the non organic honey.

Molecular detection and differentiation of Brazilian and African isolates of the entomopathogen Neozygites tanajoae (Entomophthorales: Neozygitaceae) with PCR using specific primers

Neozygites tanajoae is an entomopathogenic fungus which has been used for biocontrol of the cassava green mite (Mononychellus tanajoa, CGM) in Africa. Establishment and dispersal of Brazilian isolates which have been introduced into some African countries in recent years to improve CGM control was followed with specific PCR assays. Two primer pairs, NEOSSU_F/NEOSSU_R and 8DDC_F/8DDC_R, were used to differentiate isolates collected from several locations in Brazil and from three countries in Africa, Benin, Ghana and Tanzania. The first primer pair enabled the species-specific detection of Neozygites tanajoae, while the second differentiated the Brazilian isolates from those of other geographical origin. PCR assays were designed for detection of fungal DNA in the matrix of dead infested mites since N. tanajoae is difficult to isolate and culture on selective artificial media. Our results show that all isolates (Brazilian and African) that sporulated on mummified mites were amplified with the first primer pair confirming their Neozygites tanajoae identity. The second pair amplified DNA from all the Brazilian isolates, but did not amplify any DNA samples from the African isolates. None of the two primers showed amplification neither from any of the non-sporulating mite extracts nor from the dead uninfected mites used as negative controls. We confirmed that the two primer pairs tested are suitable for the detection and differential identification of N. tanajoae isolates from Brazil and Africa and that they are useful to monitor the establishment and spread of the Brazilian isolates of N. tanajoae introduced into Benin or into other African countries for improvement of CGM biocontrol.

Evaluation of damage caused on Coffea arabica by a population of Pratylenchus coffeae considered non-pathogenic on coffee

Evaluation of damage caused on Coffea arabica by a population of Pratylenchus coffeae considered non-pathogenic on coffee Two greenhouse experiments were carried out in order to evaluate the damage caused on Arabica coffee (Coffea arabica) by an M(2) population of Pratylenchus coffeae, apparently non-pathogenic to coffee. Experiment 1, with `Catua Vermelho` coffee at stage of two leaf pairs and with the initial nematode densities (Pi) of 0; 333; 1,000; 3.000; and 9,000 per plant, demonstrated that M(2) can damage young coffee plants, although it is unable to reproduce on coffee roots. Experiment 2, with the same coffee cultivar and nematode densities, but with plants at stage of six leaf pairs, showed that the M2 population was unable to cause damage. Therefore, it was established that M(2) is a population of P. coffeae without reproduction on Arabica coffee, which causes damage only in the first generation on young coffee below stages of six leaf pairs.

TESA-blot for the diagnosis of Chagas disease in dogs from co-endemic regions for Trypanosoma cruzi, Trypanosoma evansi and Leishmania chagasi

We standardized serodiagnosis of dogs infected with Trypanosoma cruzi using TESA (trypomastigote excreted-secreted antigen)-blot developed for human Chagas disease. TESA-blot showed 100% sensitivity and specificity. In contrast, ELISA using TESA (TESA-ELISA) or epimastigotes (epi-ELISA) as antigen yielded 100% sensitivity but specificity of 94.1% and 49.4%, respectively. When used in field studies in an endemic region for Chagas disease, visceral leishmaniasis and Trypanosoma evansi (Mato Grosso do Sul state, Central Brazil), positivities were 9.3% for TESA-blot, 10.7% for TESA-ELISA and 32% for epi-ELISA. Dogs from a non-endemic region for these infections (Rondonia state, western Amazonia) where T cruzi is enzootic showed positivity of 4.5% for TESA-blot and epi-ELISA and 6.8% for TESA-ELISA. Sera from urban dogs from Santos, Sao Paulo, where these diseases are absent, yielded negative results. TESA-blot was the only method that distinguished dogs infected with T cruzi from those infected with Leishmania chagasi and/or Trypanosoma evansi. (C) 2009 Published by Elsevier B.V.

Identification of microsatellites markers associated with yield components and quality parameters in sugarcane

Marker assisted selection depends on the identification of tightly linked association between marker and the trait of interest. In the present work, functional (EST-SSRs) and genomic (gSSRs) microsatellite markers were used to detect putative QTLs for sugarcane yield components (stalk number, diameter and height) and as well as for quality parameters (Brix, Pol and fibre) in plant cane. The mapping population (200 individuals) was derived from a bi-parental cross (IACSP95-3018 x IACSP93-3046) from the IAC Sugarcane Breeding Program. As the map is under construction, single marker trait association analysis based on the likelihood ratio test was undertaken to detect the QTLs. Of the 215 single dose markers evaluated (1:1 and 3:1), 90 (42%) were associated with putative QTLs involving 43 microsatellite primers (18 gSSRs and 25 EST-SSRs). For the yield components, 41 marker/trait associations were found: 20 for height, 6 for diameter and 15 for stalk number. An EST-SSRs marker with homology to non-phototropic hypocotyls 4 (NPH4) protein was associated with a putative QTL with positive effect for diameter as also with a negative effect for stalk number. In relation to the quality parameters, 18 marker trait associations were found for Brix, 19 for Pol, and 12 for fibre. For fibre, 58% of the QTLs detected showed a negative effect on this trait. Some makers associated with QTLs with a negative effect for fibre showed a positive effect for Pol, reflecting the negative correlation generally observed between these traits.

Antioxidant enzyme activity in Acidithiobacillus ferrooxidans LR maintained in contact with chalcopyrite

The total protein content and activity of the enzymes glutathione reductase (GR), superoxide dismutase (SOD) and thioredoxin reductase (TrxR) were evaluated in Acidithiobacillus ferrooxidans LR cells maintained in contact with the metal sulfide chalcopyrite for 1 and 10 days. A significant decrease in total protein content was observed in cells maintained for 10 days in the presence of chalcopyrite, suggesting proteolytic breakdown clue to exposure to the metal sulfide. Following 10 clays of contact with chalcopyrite, increases in GR, SOD and TrxR activities were detected, suggesting the formation of reactive oxygen species. After ten clays, there was a fivefold increase in GR activity, of which, isoenzyme IV represented approximately 82% of the total. An increase in Fe-SOD activity following ten days exposure to chalcopyrite was also determined, as measured on non-denaturing polyacrylamide gels. Also, after 10 days. an approximately 31-fold increase was observed for TrxR activity. The presence of oxidative stress when A. ferrooxidans is in the presence of chalcopyrite could have a negative impact on bioleaching. (C) 2010 Elsevier Ltd. All rights reserved.

Culturable endophytic filamentous fungi from leaves of transgenic imidazolinone-tolerant sugarcane and its non-transgenic isolines

The diversity of endophytic filamentous fungi from leaves of transgenic imidazolinone-tolerant sugarcane plants and its isoline was evaluated by cultivation followed by amplified rDNA restriction analysis (ARDRA) of randomly selected strains. Transgenic and non-transgenic cultivars and their crop management (herbicide application or manual weed control) were used to assess the possible non-target effects of genetically modified sugarcane on the fungal endophytic community. A total of 14 ARDRA haplotypes were identified in the endophytic community of sugarcane. Internal transcribed spacer (ITS) sequencing revealed a rich community represented by 12 different families from the Ascomycota phylum. Some isolates had a high sequence similarity with genera that are common endophytes in tropical climates, such as Cladosporium, Epicoccum, Fusarium, Guignardia, Pestalotiopsis and Xylaria. Analysis of molecular variance indicated that fluctuations in fungal population were related to both transgenic plants and herbicide application. While herbicide applications quickly induced transient changes in the fungal community, transgenic plants induced slower changes that were maintained over time. These results represent the first draft on composition of endophytic filamentous fungi associated with sugarcane plants. They are an important step in understanding the possible effects of transgenic plants and their crop management on the fungal endophytic community.

Performance of `Tahiti` lime on twelve rootstocks under irrigated and non-irrigated conditions

Faced with new challenges, such as emerging diseases, shortening of orchard longevity, and larger social and environmental demands from consumers, practices such as rootstock diversification, irrigation and high density plantings have become relevant for the Brazilian citrus industry. This research had the objective to evaluate the performance of irrigated and non-irrigated `Tahiti` lime trees grafted on 12 rootstocks and one interstock. Plots were distributed following a randomized block design, with four replicates and one plant per plot. Rootstocks influenced plant vigor, especially `Flying Dragon` trifoliate, which reduced tree height by approximately 47% compared to the `Rangpur lime. Trees that were budded on more vigorous rootstocks showed higher yield when grown without irrigation than with irrigation. The `1646` citradia and `Morton` citrange rootstocks performed particularly well. On the other hand, the plants on less vigorous rootstocks showed better performance in terms of yield under irrigation than the same combinations without irrigation, especially those grafted on the tetraploid `Carrizo` and `Troyer` citranges, `Swingle` citrumelo, `Davis A` trifoliate and `Flying Dragon` trifoliate. Plants budded on the `1708` citradia had high yields under irrigated and non-irrigated conditions. The effect of interstock on plant vigor was dependent of rootstock. Interstocked plants on `Davis A` trifoliate were higher than those without interstock. On the other hand, interstocked plants on Catania 2 `Volkamer` lemon were less vigorous than those without interstock. (C) 2011 Elsevier B.V. All rights reserved.

The bacterial diversity in a Brazilian non-disturbed mangrove sediment

The bacterial diversity present in sediments of a well-preserved mangrove in Ilha do Cardoso, located in the extreme south of So Paulo State coastline, Brazil, was assessed using culture-independent molecular approaches (denaturing gradient gel electrophoresis (DGGE) and analysis of 166 sequences from a clone library). The data revealed a bacterial community dominated by Alphaproteobacteria (40.36% of clones), Gammaproteobacteria (19.28% of clones) and Acidobacteria (27.71% of clones), while minor components of the assemblage were affiliated to Betaproteobacteria, Deltaproteobacteria, Firmicutes, Actinobacteria and Bacteroidetes. The clustering and redundancy analysis (RDA) based on DGGE were used to determine factors that modulate the diversity of bacterial communities in mangroves, such as depth, seasonal fluctuations, and locations over a transect area from the sea to the land. Profiles of specific DGGE gels showed that both dominant (`universal` Bacteria and Alphaproteobacteria) and low-density bacterial communities (Betaproteobacteria and Actinobacteria) are responsive to shifts in environmental factors. The location within the mangrove was determinant for all fractions of the community studied, whereas season was significant for Bacteria, Alphaproteobacteria, and Betaproteobacteria and sample depth determined the diversity of Alphaproteobacteria and Actinobacteria.

Microbial activity and community composition in saline and non-saline soils exposed to multiple drying and rewetting events

The effects of drying and rewetting (DRW) have been studied extensively in non-saline soils, but little is known about the impact of DRW in saline soils. An incubation experiment was conducted to determine the impact of 1-3 drying and re-wetting events on soil microbial activity and community composition at different levels of electrical conductivity in the saturated soil extract (ECe) (ECe 0.7, 9.3, 17.6 dS m(-1)). A non-saline sandy loam was amended with NaCl to achieve the three EC levels 21 days prior to the first DRW; wheat straw was added 7 days prior to the first DRW. Each DRW event consisted of 1 week drying and 1 week moist (50% of water holding capacity, WHC). After the last DRW, the soils were maintained moist until the end of the incubation period (63 days after addition of the wheat straw). A control was kept moist (50% of WHC) throughout the incubation period. Respiration rates on the day after rewetting were similar after the first and the second DRW, but significantly lower after the third DRW. After the first and second DRW, respiration rates were lower at EC17.6 compared to the lower EC levels, whereas salinity had little effect on respiration rates after the third DRW or at the end of the experiment when respiration rates were low. Compared to the continuously moist treatment, respiration rates were about 50% higher on day 15 (d15) and d29. On d44, respiration rates were about 50% higher at EC9.7 than at the other two EC levels. Cumulative respiration was increased by DRW only in the treatment with one DRW and only at the two lower EC levels. Salinity affected microbial biomass and community composition in the moist soils but not in the DRW treatments. At all EC levels and all sampling dates, the community composition in the continuously moist treatment differed from that in the DRW treatments, but there were no differences among the DRW treatments. Microbes in moderately saline soils may be able to utilise substrates released after multiple DRW events better than microbes in non-saline soil. However, at high EC (EC17.6), the low osmotic potential reduced microbial activity to such an extent that the microbes were not able to utilise substrate released after rewetting of dry soil.

Genomics of pyrrolnitrin biosynthetic loci: evidence for conservation and whole-operon mobility within Gram-negative bacteria

Pyrrolnitrin (PRN) is a tryptophan-derived secondary metabolite produced by a narrow range of Gram-negative bacteria. The PRN biosynthesis by rhizobacteria presumably has a key role in their life strategies and in the biocontrol of plant diseases. The biosynthetic operon that encodes the pathway that converts tryptophan to PRN is composed of four genes, prnA through D, whose diversity, genomic context and spread over bacterial genomes are poorly understood. Therefore, we launched an endeavour aimed at retrieving, by in vitro and in silico means, diverse bacteria carrying the prnABCD biosynthetic loci in their genomes. Analysis of polymorphisms of the prnD gene sequences revealed a high level of conservation between Burkholderia, Pseudomonas and Serratia spp. derived sequences. Whole-operon- and prnD-based phylogeny resulted in tree topologies that are incongruent with the taxonomic status of the evaluated strains as predicted by 16S rRNA gene phylogeny. The genomic composition of c. 20 kb DNA fragments containg the PRN operon varied in different strains. Highly conserved and distinct transposase-encoding genes surrounding the PRN biosynthetic operons of Burkholderia pseudomallei strains were found. A prnABCD-deprived genomic region in B. pseudomallei strain K96243 contained the same gene composition as, and shared high homology with, the flanking regions of the PRN operon in B. pseudomallei strains 668, 1106a and 1710b. Our results strongly suggest that the PRN biosynthetic operon is mobile. The extent, frequency and promiscuity of this mobility remain to be understood.

The effect of storage on the solubilization pattern of bean hull non-starch polysaccharides

The storage of Carioca bean at 30 C and 75% relative humidity for eight months altered the solubilization pattern of hulls non-starch polysaccharides The polysaccharide physicochemical pattern changed resulting in a shift in the composition of water-soluble and water-insoluble polysaccharides caused by the insolubilization of galacturonans and xyloglucan Hulls make up 10% of whole beans which showed an increase of about 5% in water-insoluble polysaccharides and a decrease of about 1% in water-soluble polysaccharides with aging These values suggest that cotyledons and hulls together account for an increase of about 2 g of water-insoluble polysaccharides and a decrease of 1 5 g of water-soluble polysaccharides per 100 g of beans This change in the polysaccharide composition may produce a considerable difference in the dietary fiber profile The alterations observed in bean hull non-starch polysaccharide composition were similar to those previously observed in the cotyledon (C) 2010 Elsevier Ltd All rights reserved

Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits from Brazil

The bioactive compounds and antioxidant capacities of polyphenolic extracts of 18 fresh and dry native non-traditional fruits from Brazil were determined using ABTS, DDPH, FRAP and beta-carotene bleaching methods. The study provides an adaptation of these methods, along with an evaluation of the compounds related to antioxidant potential. The results show promising perspectives for the exploitation of non-traditional tropical fruit species with considerable levels of nutrients and antioxidant capacity. Although evaluation methods and results reported have not yet been sufficiently standardised, making comparisons difficult, our data add valuable information to current knowledge of the nutritional properties of tropical fruits, such as the considerable antioxidant capacity found for acerola - Malpighia emarginata and camu-camu - Myrciaria dubia (ABTS, DPPH and FRAP) and for puca-preto - Mouriri pusa (all methods). (C) 2010 Elsevier Ltd. All rights reserved.

Effect of cooking on non-starch polysaccharides of hard-to-cook beans

Experiments carried out to study changes induced by hard-to-cook (HTC) phenomenon in the non-starch polysaccharides of beans stored at 30 degrees C and 75% RH for 8 months showed that the development of HTC did not affect the amounts of soluble and insoluble fibre in cooked seeds but changed their carbohydrates physical properties. Aged beans non-starch polysaccharides presented lower water-solubility and underwent lower degradation of galacturonans and arabinose-rich polysaccharides when submitted to cooking. The decrease in non-starch polysaccharides water-solubility produced a shift in the polymers fractionation profile which resulted in an increase of weak and middle-alkali soluble polymers bulk as well as in their arabinose and uronic acid contents. Uronic acid contents were higher in polymers released by 1 M NaOH and in the cellulose-rich residues while the arabinose contents were higher in the mild-alkali soluble polymers of aged seeds. Methylation analysis showed no evident alterations in the xyloglucans and arabinans branching degree with beans ageing. However, both, the molecular mass of water-soluble pectins and CDTA-soluble pectins, increased. Even though changes in the non-starch polysaccharide solubility were not related to the decrease in the arabinan and xyloglucan degree of branching they may be related to the formation of new chemical interactions other than hydrogen bond. There was a correlation between acidic and neutral polysaccharides insolubilisation in beans ageing and probably in beans hardening. After processing, aged seeds present higher amounts of insoluble fibre when compared to normal beans. (C) 2008 Elsevier Ltd. All rights reserved.

Non-starch polysaccharide composition of two cultivars of banana (Musa acuminata L.: cvs Mysore and Nanicao)

Fruits represent a rich source of soluble and insoluble fibre, and the pectin is the most common and known soluble fraction from the cell wall solubilization occurring during fruit ripening. Banana fruit, for example, is one of the most consumed fruits in the world, but its non-starch polysaccharide composition is almost unknown. Despite few works have been carried out about the enzymes concerning cell wall loosening focusing banana ripening, there is no knowledge about the composition of the banana cell wall. Moreover, there is no information about the influence of the cultivar in that composition. Nanicao and Mysore cultivars were chosen for this work because of their differential accumulation of both starch during development and amounts of total fibre in the ripe fruit. Nanicao and Mysore had their fibres subfractioned and their composition analysed. Results showed that the cultivars are distinct not only in terms of starch and soluble sugars accumulation, but also in non-starch polysaccharides amounts and composition. Non-starch polysaccharides are similar in total amounts in both banana cultivars (similar to 3.5), but substantially different in the content of CDTA and NaOH-4M soluble fractions and also in the molecular mass distribution of WSP and CDTA. Nanicao has more calcium-linked pectin than Mysore, which in turn is richer in hemicellulose-like polysaccharides. Both cultivars likewise cereals polysaccharides seem to be composed of galacturonans and arabinoxylans.(c) 2007 Elsevier Ltd. All rights reserved.