Molecular divergence in the timeless and cpr genes among three sympatric cryptic species of the Anopheles triannulatus complex

Anopheles triannulatus s.l. is a malaria vector with a wide geographic distribution, ranging from Argentina-Nicaragua and Trinidad. Here we analysed sequences of two genes, timeless and cpr, to assess the genetic variability and divergence among three sympatric cryptic species of this complex from Salobra, central-western Brazil. The timeless gene sequences did not conclusively differentiate Anopheles halophylus and An. triannulatus species "C". However, a partial separation has been observed between these species and An. triannulatus s.s. Importantly, the analysis of the cpr gene sequences revealed fixed differences, no shared polymorphisms and considerable genetic differentiation among the three species of the An. triannulatus complex. The results confirm that An. triannulatus s.s., An. halophylus and An. triannulatus species C are distinct taxa, with the latter two likely representing a more recent speciation event.

First detection of Rio Negro virus (Venezuelan equine encephalitis complex subtype VI) in Córdoba, Argentina

Rio Negro virus (RNV) (Venezuelan equine encephalitis subtype VI) circulates only in Argentina; in northern provinces, isolates have been obtained from mosquitoes and rodents since 1980 and have been associated with acute febrile illness in humans. However, no studies of RNV have been performed in the central area of the country. We carried out molecular and serological detection of RNV in Córdoba, a province of the central part of the country, in mosquitoes and humans, respectively. One mosquito pool tested positive for alphavirus RNA by reverse transcriptase-nested polymerase chain reaction (RT-nested PCR). Subsequent sequencing determined that this alphavirus grouped with RNV. Serological studies detected antibodies to RNV in one human serum sample, which was obtained during the same period that RNV was detected using the aforementioned molecular methods. This is the first report of RNV circulation in the central area of Argentina, indicating an expansion of its original distribution. These results highlight the importance of strengthening surveillance procedures in endemic areas, as well as in new regions where RNV may emerge.

Revalidation and redescription of Triatoma brasiliensis macromelasoma Galvão, 1956 and an identification key for the Triatoma brasiliensis complex (Hemiptera: Reduviidae: Triatominae)

Triatoma brasiliensis macromelasoma is revalidated based on the results of previous multidisciplinary studies on the Triatoma brasiliensis complex, consisting of crossing experiments and morphological, biological, ecological and molecular analyses. These taxonomic tools showed the closest relationship between T. b. macromelasoma and Triatoma brasiliensis brasiliensis. T. b. macromelasoma is redescribed based on specimens collected in the type locality and specimens from a F1 colony. The complex now comprises T. b. brasiliensis, T. b. macromelasoma, Triatoma melanica, Triatoma juazeirensis and Triatoma sherlocki. An identification key for all members of the complex is presented. This detailed comparative study of the morphological features of T. b. macromelasoma and the remaining members of the complex corroborates results from multidisciplinary analyses, suggesting that the subspecific status is applicable. This subspecies can be distinguished by the following combination of features: a pronotum with 1+1 narrow brownish-yellow stripes on the submedian carinae, not attaining its apex, hemelytra with membrane cells darkened on the central portion and legs with an incomplete brownish-yellow ring on the apical half of the femora. Because the T. brasiliensis complex is of distinct epidemiological importance throughout its geographic distribution, a precise identification of its five members is important for monitoring and controlling actions against Chagas disease transmission.

The identification and differentiation of the Candida parapsilosis complex species by polymerase chain reaction-restriction fragment length polymorphism of the internal transcribed spacer region of the rDNA

Currently, it is accepted that there are three species that were formerly grouped under Candida parapsilosis: C. para- psilosis sensu stricto, Candida orthopsilosis, andCandida metapsilosis. In fact, the antifungal susceptibility profiles and distinct virulence attributes demonstrate the differences in these nosocomial pathogens. An accurate, fast, and economical identification of fungal species has been the main goal in mycology. In the present study, we searched sequences that were available in the GenBank database in order to identify the complete sequence for the internal transcribed spacer (ITS)1-5.8S-ITS2 region, which is comprised of the forward and reverse primers ITS1 and ITS4. Subsequently, an in silico polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to differentiate the C. parapsilosis complex species. Ninety-eight clinical isolates from patients with fungaemia were submitted for analysis, where 59 isolates were identified as C. parapsilosis sensu stricto, 37 were identified as C. orthopsilosis, and two were identified as C. metapsilosis. PCR-RFLP quickly and accurately identified C. parapsilosis complex species, making this method an alternative and routine identification system for use in clinical mycology laboratories.

Anopheles goeldii Rozeboom & Gabaldón (Diptera, Culicidae): a species of the Nuneztovari Complex of Anopheles Meigen

Anopheles (Nyssorhynchus) goeldiiRozeboom & Gabaldón, 1941, a species of the Nuneztovari Complex, was described based on morphological characteristics of the male, female, larva, pupa, and eggs. The type locality is Boa Vista (= Fordlândia), a district in the vicinity of Rio Tapajós, in the municipality of Aveiro, in the state of Pará, Brazil. Anopheles goeldii is redescribed based on morphological traits of the fourth instar larva, pupa, egg, and male and female. DNA sequences from the cytochrome oxidase subunit I (COI barcode region) of the mitochondrial genome were utilized for species characterization. Specimens of An. goeldii from the Pará, Amapá, and Amazonas states were employed to redescribe the species and to compare with morphologically similar taxa.

Comparison between detailed digital and conventional soil maps of an area with complex geology

Since different pedologists will draw different soil maps of a same area, it is important to compare the differences between mapping by specialists and mapping techniques, as for example currently intensively discussed Digital Soil Mapping. Four detailed soil maps (scale 1:10.000) of a 182-ha sugarcane farm in the county of Rafard, São Paulo State, Brazil, were compared. The area has a large variation of soil formation factors. The maps were drawn independently by four soil scientists and compared with a fifth map obtained by a digital soil mapping technique. All pedologists were given the same set of information. As many field expeditions and soil pits as required by each surveyor were provided to define the mapping units (MUs). For the Digital Soil Map (DSM), spectral data were extracted from Landsat 5 Thematic Mapper (TM) imagery as well as six terrain attributes from the topographic map of the area. These data were summarized by principal component analysis to generate the map designs of groups through Fuzzy K-means clustering. Field observations were made to identify the soils in the MUs and classify them according to the Brazilian Soil Classification System (BSCS). To compare the conventional and digital (DSM) soil maps, they were crossed pairwise to generate confusion matrices that were mapped. The categorical analysis at each classification level of the BSCS showed that the agreement between the maps decreased towards the lower levels of classification and the great influence of the surveyor on both the mapping and definition of MUs in the soil map. The average correspondence between the conventional and DSM maps was similar. Therefore, the method used to obtain the DSM yielded similar results to those obtained by the conventional technique, while providing additional information about the landscape of each soil, useful for applications in future surveys of similar areas.

Contaminação endógena por Aspergillus spp. em milho pós-colheita no Estado do Paraná

Sessenta amostras de milho pós-colheita foram avaliadas quanto à contaminação fúngica endógena e o potencial toxígeno de espécies do gênero Aspergillus e seus teleomorfos. Quarenta grãos aparentemente sadios de cada amostra foram desinfestados em NaClO e incubados em câmara úmida a 25±1ºC para exteriorização dos fungos, que posteriormente foram isolados em ágar Czapek-Dox. Foram identificadas as espécies Aspergillus flavus, A. parasiticus, Eurotium amstelodami e E. chevalieri. O potencial toxígeno dos fungos A. flavus e A. parasiticus foi avaliado quanto à síntese de aflatoxinas em meio ágar-coco. Espécies do gênero Eurotium foram avaliadas quanto à síntese de esterigmatocistina, nos meios ágar-amendoim e trigo triturado. A porcentagem de grãos contaminados variou entre 0 e 100%, prevalecendo os gêneros Aspergillus, Penicillium e Fusarium. A espécie predominante foi a A. flavus (64%), seguida por E. amstelodami (19%), E. chevalieri (10%) e A. parasiticus (7%). A partir de 109 isolados de A. flavus, evidenciou-se que 73 isolados sintetizaram aflatoxinas B1 e B2, 20 sintetizaram B1, sete sintetizaram B1 e G1, três sintetizaram B1, B2 e G1 e em seis não foi detectada a síntese de aflatoxina. A síntese de esterigmatocistina pelas espécies E. amstelodami e E. chevalieri não foi detectada.

Inter and intra-guild interactions in egg parasitoid species of the soybean stink bug complex

The objective of this research was to evaluate the parasitism behavior of Telenomus podisi Ashmead, Trissolcus basalis (Wollaston) e Trissolcus urichi Crawford (Hymenoptera: Scelionidae) on eggs of Nezara viridula L., Euschistus heros F., Piezodorus guildinii Westwood and Acrosternum aseadum Rolston (Heteroptera: Pentatomidae), in no choice and multiple choice experiments. For all parasitoid species, the results demonstrated the existence of a main host species that maximizes the reproductive success. The competitive interactions among the parasitoid species were investigated in experiments of sequential and simultaneous release of different combinations of parasitoid pairs on the hosts N. viridula, E. heros and A. aseadum. Exploitative competition was observed for egg batches at the genus level (Telenomus vs. Trissolcus) and interference competition at the species level (T. basalis vs. T. urichi). Trissolcus urichi was the most aggressive species, interfering with the parasitism of T. basalis. Generally, T. basalis showed an opportunistic behavior trying to parasitise eggs after T. urichi had abandoned the egg batch. The selection of parasitoid species for use in augmentative biological control programs should take into account the diversity of pentatomids present in soybean in addition to the interactions among the different species of parasitoids.

New species emergence via recombination among isolates of the Brazilian tomato infecting Begomovirus complex

Partial nucleotide sequences of five tomato infecting Begomovirus isolates were determined from DNA-A fragments, corresponding to the 5' region of the replication associated protein gene, the intergenic region and the 5' region of the coat protein gene. Isolate DFM shared 95% identity with Tomato mottle leaf curl virus (TMoLCV), isolates 34, PA-05, and Ta4 were 88% identical to Tomato yellow vein streak virus and isolate DF-BR3 shared 77% identity with TMoLCV. Recombination analysis indicated that isolate DF-BR3 was a chimaera, and it provided evidence that there is a complex and actively recombining population of tomato infecting begomoviruses in Brazil.

Utilização de penas de galinha para produção de queratinase por Aspergillus carbonarius

O objetivo deste trabalho foi estudar a produção de queratinase por Aspergillus carbonarius URM 1546, tendo-se como substrato penas de galinha, por meio de planejamento fatorial completo 2³. Todos os parâmetros estudados e as interações de segunda ordem foram estatisticamente significativos. A maior atividade queratinolítica (48,9 U mL-1) foi obtida com 120 rpm, 0,5% (p/v) de penas de galinha e sete dias de cultivo.

Variáveis que influenciam a produção de celulases e xilanase por espécies de Aspergillus

O objetivo deste trabalho foi determinar as variáveis que influenciam a produção simultânea de celulases e xilanase por Aspergillus aculeatus URM 4953 e A. phoenicis URM 4924, com uso de bagaço de cana-de-açúcar como substrato. As variáveis avaliadas foram pH inicial, tempo de cultivo, concentração do substrato, agitação, concentração do inóculo, temperatura, tipo de bagaço de cana-de-açúcar e espécie de Aspergillus. Foi utilizado o planejamento fatorial fracionário 2(8-4), com três repetições no ponto central. Todas as variáveis analisadas influenciaram a produção de pelo menos uma das enzimas. As maiores atividades enzimáticas observadas foram: celulases totais, 0,45 UI mL-1; endoglucanase, 0,60 UI mL-1; exoglucanase, 0,17 U mL-1 ; celobiase 6,42 UI mL-1; e xilanase, 30,05 U mL-1. As melhores condições para produção das enzimas foram: pH 6,0; tempo de cultivo de 168 horas; bagaço de cana-de-açúcar residual como substrato; e A. aculeatus URM 4953, que produziu as celulases e a xilanase simultaneamente. As variáveis que influenciam a produção simultânea das celulases e xilanase são pH, tempo de cultivo, tipo de bagaço de cana-de-açúcar e espécie de Aspergillus.

Produção de celulases por Aspergillus niger por fermentação em estado sólido

O objetivo deste trabalho foi avaliar a produção enzimática de celulases pelo fungo filamentoso Aspergillus niger por fermentação em estado sólido de diferentes substratos. Foram avaliados os substratos sólidos bagaço de cana-de-açúcar, farelo de soja, farelo de trigo e misturas entre os substratos. Em substrato com 90% de bagaço e 10% de farelo de soja, avaliaram-se os efeitos do conteúdo de umidade (60, 70 e 80%, base úmida) e da suplementação com os meios indutores da atividade enzimática: sacarose, Mandels & Weber básico, Mandels & Weber modificado, com acréscimo de carboximetilcelulose, e Czapeck Dox. As maiores atividades de celulase total e endoglucanase, em farelo de trigo, foram obtidas após 72 horas de cultivo: 0,4 e 21,0 UI g-1, respectivamente. Observou-se expressivo aumento nas atividades enzimáticas na medida em que se aumentou a proporção de farelos no substrato, em comparação à fermentação com bagaço de cana apenas. O conteúdo de umidade de 50% foi insuficiente para conseguir completa hidratação do bagaço de cana, e a umidade ideal varia de acordo com o meio utilizado para suplementação e encontra-se entre 70 e 80%. O meio de Mandels & Weber modificado apresenta o melhor resultado como indutor da atividade enzimática.

Improvement of productivity and polysaccharide-protein complex in Agaricus blazei

The objective of this work was to assess the productivity and polysaccharide-protein complex content of Agaricus blazei on rice straw medium, in comparison to conventional sawdust, using four casing soils. The A. blazei strain used was BCRC36814T, purchased from the Food Industry Research and Development Institute, Hsin-Chu, Taiwan. The two media were evaluated as to A. blazei productivity, harvesting time, and production costs. The experimental design used was a randomized complete block, with four replicates. Three local casing soils - Typic Paleudult (CCe), Typic Udorthent (Tq) and Oxyaquic Paleudult (TSp) - were compared to imported peat soil (PS, Saprists, Histosols), used as the control. The productivity of A. blazei using Tq and TSp soil was significantly higher. The TSp casing treatment resulted in earlier harvest by at least 14 to 27 days, when compared to the other treatments. The polysaccharide content in CCe (13.2%) and Tq soils (13.2%) did not differ significantly from the PS (13.4%) and TSp (10.6%) treatments. Local casing soils decreased the production costs of A. blazei cultivation. Composted rice straw can substitute sawdust as the culture medium for A. blazei production with increased yield.

Assessment of intima-media complex in carotid, femoral and right subclavian arteries for early investigation of atherosclerosis in HIV-infected patients

Objective To compare automatic and manual measurements of intima-media complex (IMC) in common carotid, common femoral and right subclavian arteries of HIV-infected patients in relation to a control group, taking into consideration the classical risk factors for atherosclerosis. Materials and Methods The study sample comprised 70 HIV-infected patients and 70 non-HIV-infected controls paired according sex and age. Automatic (gold standard) and manual measurements of IMC were performed in the carotid arteries. Manual measurements were also performed in common femoral and right subclavian arteries. Bland-Altman graphs were utilized in the comparison and the adopted level significance was 5%. Results Intima-media complex alterations were not observed in any of the individuals as the mean automatic measurement in the right common carotid (RCC) artery was considered as the gold standard. As the gold standard was compared with the manual measurements (mean, maximum and minimum), no clinically significant alteration was observed. As the gold standard was compared with other sites, the difference was statistically and clinically significant at the origin of right subclavian artery (RCC: 0.51 mm vs. 0.91 mm) (p < 0.001). Conclusion HIV-infected individuals are not at higher risk for atherosclerosis than the control population.