Sexual dimorphism in Kinosternon scorpioides (Linnaeus, 1766) from the Brazilian Amazon.

The sexual dimorphism of the Kinosternon scorpioides was evaluated using two different techniques (linear and geometric morphometry) from images and linear measurements of the carapace and plastron of adults (male and female). Linear morphometry indicated that the height and width of carapace and plastron are statistically different between sexes, with females being wider and taller. In the evaluation of geometric morphometry, ANOVA demonstrated variation in the size of the plastron and the shape of the carapace and plastron, expressing a tendency in shape for each sex. Sexual dimorphism, therefore, is verified for this species, notably by the plastron. This study indicates an additional tool for the phenotypic knowledge of animals, contributing to the study of threatened populations.

Addition of a Second Metal (Co) to Molybdenum Carbide: Effect of the Doping Route.

Molybdenum carbide is an interesting and versatile material, which has important applications in the metal matrix industry as a reinforcement material, as well as in the catalytic field. Though many papers suggest different methodologies for adding cobalt to the carbide structure aiming either to increase catalytic activity or enhancing mechanical proprieties such as ductility, etc. no straightforward evaluation is available. In the present paper two doping methodologies were studied: via solid state mixture of powders and via wet impregnation. Ammonium molybdate [(NH4)2MoO4] and cobalt nitrate [Co(NO3)2·6H2O] were used as starting materials and the doping process was carried out before carburization reaction. Those materials were characterized by FT-IR, SEM, XRF and XRD. The carbo-reduction products' were evaluated on XRD and XRF basis. Doped precursors' evaluation showed that the wet impregnated doped materials presented smaller particle sizes, were more homogeneous and retained more cobalt than the solid state doped ones. However, final products' assessment indicated that the solid state methodology was able to retain a greater dopant percentage according to XRF evaluation, and XRD data indicated a more intrinsic addition of the dopant to the carbide structure. In addition, no significant changes on particle size could be attributed to any of the methodologies, both producing Mo2C of approximately 30 nm.

Perfil clínico e microbiológico de cães com e sem otoacaríase / Clinical and microbiological profile of dogs with and without otoacariasis

Os objetivos do estudo foram identificar a presença de microrganismos nos condutos auditivos dos cães através dos exames citológico e microbiológico, assim como avaliar a associação destes à otoacaríase e seus sinais clínicos. O diagnóstico da infestação por Otodectes cynotis foi realizado através de otoscopia bilateral e pela coleta de material e visualização do parasito sob microscópio esterioscópico, constituindo o exame parasitológico. Outras coletas de secreção otológica de cada orelha foram realizadas para confecção de lâminas para a citologia e para o isolamento microbiológico, sendo este último material coletado através de "swab" estéril. Dos 250 animais examinados, em 15 (6%) foi identificado o ácaro O. cynotis.Observou-se associação entre a presença do parasito e a ocorrência de otite clínica externa caracterizada especialmente por eritema auricular (P<0,001) e a produção excessiva de secreção otológica (P=0,0016), assim como a observação, pelos proprietários, de prurido ótico nos cães (P<0,001). Sugere-se a possibilidade de essas associações serem efeito da infestação. Houve também a associação (P<0,01) entre a otoacaríase e a ocorrência de microrganismos nos dois condutos auditivos de cada animal, detectados através da citologia. No entanto, não houve associação (P=0,04 e p=0,07) entre a presença do ácaro O. cynotis e a ocorrência de microrganismos no exame microbiológico das orelhas direita e esquerda, respectivamente.

The goals of this study were to identify microorganisms in dogs' ear canals through cytological and microbiological examination, and evaluate their association with otoacariasis and its clinical signs. Otodectes cynotis infestation diagnosis was achieved by bilateral otoscopy, and by parasite viewing on cerumem under stereoscopic microscope, representing the parasitological exam. Ear discharge was also collected from each canal to fix cytology slides and for microbiological isolation, but this last sample was collected with sterile swab. Among the 250 studied animals, 15 (6%) had ear mites. There was an association between the parasite and external clinical otitis specially as established by auricular erythema (P<0,001) and excessive production of ear discharge (P=0,0016), as well as ear pruritus observed by the owners (P<0,001). It has been suggested that these associations can be an effect of the infestations. There was also an association (P<0,01) between otoacariasis and microorganisms detected by cytological exam in both ear canals of each dog. However, there was no association (P=0,04 e P=0,07) between the presence of ear mite and microorganisms detected by microbiological exam of right and left ears.

Descriptor selection for banana accessions based on univariate and multivariate analysis.

Our objective was to establish a minimum number of morphological descriptors for the characterization of banana germplasm and evaluate the efficiency of removal of redundant characters, based on univariate and multivariate statistical analyses. Phenotypic characterization was made of 77 accessions from Bahia, Brazil, using 92 descriptors. The selection of the descriptors was carried out by principal components analysis (quantitative) and by entropy (multi-category). Efficiency of elimination was analyzed by a comparative study between the clusters formed, taking into consideration all 92 descriptors and smaller groups. The selected descriptors were analyzed with the Ward-MLM procedure and a combined matrix formed by the Gower algorithm. We were able to reduce the number of descriptors used for characterizing the banana germplasm (42%). The correlation between the matrices considering the 92 descriptors and the selected ones was 0.82, showing that the reduction in the number of descriptors did not influence estimation of genetic variability between the banana accessions. We conclude that removing these descriptors caused no loss of information, considering the groups formed from pre-established criteria, including subgroup/subspecies.

Diversity of somatic coliphages in coastal regions with different levels of anthropogenic activity in São Paulo State, Brazil.

Bacteriophages are the most abundant and genetically diverse viruses on Earth, with complex ecology in both quantitative and qualitative terms. Somatic coliphages (SC) have been reported to be good indicators of fecal pollution in seawater. This study focused on determining the concentration of SC and their diversity by electron microscopy of seawater, plankton, and bivalve samples collected at three coastal regions in São Paulo, Brazil. The SC counts varied from <1 to 3.4 × 10(3) PFU/100 ml in seawater (73 samples tested), from <1 to 4.7 × 10(2) PFU/g in plankton (46 samples tested), and from <1 to 2.2 × 10(1) PFU/g in bivalves (11 samples tested). In seawater samples, a relationship between the thermotolerant coliforms and Escherichia coli and SC was observed at the three regions (P = 0.0001) according to the anthropogenic activities present at each region. However, SC were found in plankton samples from three regions: Baixada Santista (17/20), Canal de São Sebastião (6/14), and Ubatuba (3/12). In seawater samples collected from Baixada Santista, four morphotypes were observed: A1 (4.5%), B1 (50%), C1 (36.4%), and D1 (9.1%). One coliphage, Siphoviridae type T1, had the longest tail: between 939 and 995 nm. In plankton samples, Siphoviridae (65.8%), Podoviridae (15.8%), Microviridae (15.8%), and Myoviridae (2.6%) were found. In bivalves, only the morphotype B1 was observed. These SC were associated with enteric hosts: enterobacteria, E. coli, Proteus, Salmonella, and Yersinia. Baixada Santista is an area containing a high level of fecal pollution compared to those in the Canal de São Sebastião and Ubatuba. This is the first report of coliphage diversity in seawater, plankton, and bivalve samples collected from São Paulo coastal regions. A better characterization of SC diversity in coastal environments will help with the management and evaluation of the microbiological risks for recreation, seafood cultivation, and consumption.

Mutation in intron 5 of GTP cyclohydrolase 1 gene causes dopa-responsive dystonia (Segawa syndrome) in a Brazilian family.

Dopa-responsive dystonia (DRD), also known as Segawa syndrome or hereditary progressive dystonia with diurnal fluctuation, is clinically characterized by the occurrence of simultaneous or late Parkinsonism and by an excellent response to treatment with low doses of L-dopa. Diagnosis of DRD is essentially clinical. It is based on clinical history and the response to treatment with low doses of L-dopa. However, due to the low penetrance of the disease, asymptomatic carriers may exist. In these cases, mutational analysis of the GCH1 gene is an alternative to diagnose DRD. In the present study, we investigated a large DRD-carrier family in an attempt to identify the disease-causing mutation. The proband, a young woman diagnosed at the age of 13 years, is the daughter of a healthy non-consanguineous couple with history of several cases, on the maternal side of the family, of tip-toeing, disturbance of gait, Parkinsonism, rigidity and cramps in the lower limbs. Using single strand conformational polymorphism and DNA sequencing techniques to analyze DNA extracted from blood samples, we identified a mutation in the GCH1 gene, IVS5+3insT, which would preclude the formation of the active enzyme due to the formation of truncated peptides.

Occurrence of Struthiopterolichus bicaudatus (Acari; Pterolichidae) in Southeastern Brazil

A ocorrência de Struthiopterolichus bicaudatus é assinalada em uma criação de avestruz no estado de Minas Gerais. Os principais sinais associados com a presença dos ácaros foram penas quebradas com a falta de barbas no vexilo, principalmente nas penas das asas, e prurido. Esses sinais foram observados em todas as aves do plantel (285 animais) com idade acima de 13 meses. As aves com idade abaixo de 13 meses (60 animais) não apresentaram qualquer sinal, tampouco ácaros.

Defect analysis in natural quartz from Brazilian sites for ionising radiation dosimetry.

The impurity-related point defects and the dosimetry properties of quartz irradiated with gamma- and X-rays doses were investigated for natural crystals taken from different geologies. The specimens were initially irradiated with gamma particles from (137)Cs to determine the sensitivity and repeatability of thermoluminescence (TL) emission at approximately 503 K. The dose response was investigated in the range 1-20 mGy. The energy dependence was considered in the range 16-65 keV with X rays and with gamma rays from 137Cs and 60Co. It was found that quartz material from two geologies exhibit a linear relationship between TL intensities and absorbed doses with high angular coefficients. Its sensitivities are higher than that found for LiF TLD-100 dosemeters irradiated in the same conditions. The results were discussed in relation to pre-existing impurity contents and the formation of Al-hole centres.

Populational structure of Schistosoma mansoni assessed by DNA microsatellites.

DNA microsatellites were used as molecular markers to analyse the population structure of the laboratory LE strain and of 10 field isolates of Schistosoma mansoni, the aetiologic agent of schistosomiasis. Out of 16,000 DNA sequences analysed in databases, 622 microsatellite loci were identified in 481 sequences (3.0%). The AT repetitions were the most frequent, followed by AAT and AC. Six loci showing perfect repetitions were selected and used in the polymerase chain reaction to evaluate polymorphisms in the number of repeats. Two groups of worms were studied. The first group consisted of 78 individuals, 39 of each sex, of the LE strain. The second group of worms consisted of 10 field isolates: seven from humans and three from snails. Four of the six loci were polymorphic, containing 11-17 alleles per locus. No linkage disequilibrium was observed among loci and none of the loci was sex linked. In both groups of worms, a significant deviation from Hardy-Weinberg equilibrium was observed. The observed heterozygosity was always lower than the expected one. The polymerase chain reaction primers were S. mansoni specific. The LE strain showed a lower total number of alleles or a lower average number of alleles/polymorphic locus than the field isolates, suggesting that 41 years of laboratory maintenance exerted selective pressure on the LE strain. The S. mansoni populations from the field were most genetically undifferentiated (R(ST)<0.027), suggesting a high gene flow among them. Our results showed the usefulness of microsatellites for population analysis of S. mansoni, offering a new alternative for a better understanding of schistosomiasis epidemiology.

Evolutionary relationships and biogeography of Biomphalaria (Gastropoda: Planorbidae) with implications regarding its role as host of the human bloodfluke, Schistosoma mansoni.

The wide geographic distribution of Schistosoma mansoni, a digenetic trematode and parasite of humans, is determined by the occurrence of its intermediate hosts, freshwater snails of the genus Biomphalaria (Preston 1910). We present phylogenetic analyses of 23 species of Biomphalaria, 16 Neotropical and seven African, including the most important schistosome hosts, using partial mitochondrial ribosomal 16S and complete nuclear ribosomal ITS1 and ITS2 nucleotide sequences. A dramatically better resolution was obtained by combining the data sets as opposed to analyzing each separately, indicating that there is additive congruent signal in each data set. Neotropical species are basal, and all African species are derived, suggesting an American origin for the genus. We confirm that a proto-Biomphalaria glabrata gave rise to all African species through a trans-Atlantic colonization of Africa. In addition, genetic distances among African species are smaller compared with those among Neotropical species, indicating a more recent origin. There are two species-rich clades, one African with B. glabrata as its base, and the other Neotropical. Within the African clade, a wide-ranging tropical savannah species, B. pfeifferi, and a Nilotic species complex, have both colonized Rift Valley lakes and produced endemic lacustrine forms. Within the Neotropical clade, two newly acquired natural hosts for S. mansoni (B. straminea and B. tenagophila) are not the closest relatives of each other, suggesting two separate acquisition events. Basal to these two species-rich clades are several Neotropical lineages with large genetic distances between them, indicating multiple lineages within the genus. Interesting patterns occur regarding schistosome susceptibility: (1) the most susceptible hosts belong to a single clade, comprising B. glabrata and the African species, (2) several susceptible Neotropical species are sister groups to apparently refractory species, and (3) some basal lineages are susceptible. These patterns suggest the existence of both inherent susceptibility and resistance, but also underscore the ability of S. mansoni to adapt to and acquire previously unsusceptible species as hosts. Biomphalaria schrammi appears to be distantly related to other Biomphalaria as well as to Helisoma, and may represent a separate or intermediate lineage.

Possible competitive displacement of planorbids by Melanoides tuberculata in Minas Gerais, Brazil.

Several species of snails, including Pomacea haustrum, Marisa cornuarietis and Helisoma duryi, have been identified as probable competitors and/or predators of planorbid intermediate hosts of Schistosoma. During the last few years, studies carried out in the Caribbean region have shown reductions and even disappearances of populations of Biomphalaria glabrata and B. straminea in breeding places where the snail Melanoides tuberculata was introduced. Observations made over a period of 10 years in two lakes close to Belo Horizonte, MG, showed that there were marked reductions in autochthonous populations of B. glabrata and B. straminea after the arrival of M. tuberculata, both Biomphalaria species disappearing completely after eight years.

Widespread heteroplasmy in schistosomes makes an mtVNTR marker "nearsighted".

Mitochondrial markers are often hailed as the preferred DNA elements for analyses of population subdivision. To this end we have employed a mitochondrial repeat element to examine the population structure in Schistosoma mansoni (human blood flukes). Schistosome isolates were collected from each of 21 different patients representing seven different areas of a Brazilian village. These parasite isolates demonstrate substantial genetic polymorphism, with an average of 10 genotypes infecting each patient, which is more readily detected because of high levels of heteroplasmy (i.e., 72.5% of the individual worms exhibit multiple versions of this repeat region with different numbers of repeats). Due to the high number of common haplotypes in the population, this repeat element from S. mansoni has a large proportion (47%) of its genetic variation described by differences among mitochondrial genomes within individual worms. However, when only rare haplotypes are considered, population structure can be detected. It seems that heteroplasmy in the schistosome population of Melquiades is both the source of plentiful genetic variation and a confounding factor in the analysis of that variation. Thus the schistosome population in Melquiades may actually be more strongly subdivided than we are able to detect using this mitochondrial marker.

The control of the schistosome-transmitting snail Biomphalaria glabrata by the plant Molluscicide Euphorbia splendens var. hislopii (syn milli Des. Moul): a longitudinal field study in an endemic area in Brazil.

Under laboratory conditions, latex from Euphorbia splendens has shown promise as a plant molluscicide for control of Biomphalaria species, intermediate hosts for Schistosoma mansoni. The purpose of this study was to evaluate its efficiency under field conditions. Application of filtered latex at 12 ppm to one stream in an endemic rural area in Minas Gerais state, Brazil, in September 1995, did result in a reduction in snail density as compared to an untreated stream but the snail population recovered quickly. However, two applications with a two-week interval of unfiltered E. splendens latex at 5 ppm in November 1996 in the same stream resulted in complete disappearance of B. glabrata and snails did not reappear until the 14th month after the applications. In the control stream, without treatment, the snails were found during all months. Laboratory studies confirmed that unfiltered latex is a more potent molluscicide than filtered latex. Considering the advantages of the latex such as its low toxicity to other aquatic animals and its photobiodegradability, as well as the simple method of application, this natural product is promising as an effective molluscicide.

Geographical distribution of Biomphalaria snails in the State of Minas Gerais, Brazil.

Published and unpublished observations on geographical distribution of Biomphalaria snails in the State of Minas Gerais, Brazil, were compiled. This work is aimed at knowing the present occurrence of Biomphalaria species in this region, and at contributing to the elaboration of the planorbid chart of Minas Gerais. In malacological surveys, performed by several researchers, the presence of seven species of this genus was recorded. Those planorbids were found in 12 mesoregions, in 283 (33.1%) municipalities out of 853 with the following distribution: B. glabrata (185 municipalities), B. straminea (125), B. tenagophila (58), B. peregrina (57), B. schrammi (26), B. intermedia (20) and B. occidentalis (2). B. glabrata and B. tenagophila are found naturally infected by Schistosoma mansoni in Minas Gerais. In 24 municipalities the three snail hosts of S. mansoni in Brazil, B. glabrata, B. tenagophila and B. straminea, are present.

Resistance of Biomphalaria occidentalis from Varzea das Flores dam, Minas Gerais, to Schistosoma mansoni infection detected by low stringency polymerase chain reaction.

Biomphalaria occidentalis Paraense, 1981 from Varzea das Flores dam, MG, Brazil, was exposed to infection with Schistosoma mansoni. Individual infection was performed with 140 B. occidentalis and 100 B. glabrata snails using LE and SJ strains. Two groups of B. occidentalis were killed after seven day-miracidia exposure to detect S. mansoni DNA, through the low stringency polymerase chain reaction (LS-PCR), and were negative. The infection rates were 69.2% (LE strain) and 96.7% (SJ strain) for B. glabrata and 0% for B. occidentalis. LS-PCR enabled early resistance diagnosis.

Biparental mitochondrial DNA inheritance in the parasitic trematode Schistosoma mansoni.

The maternal inheritance of mitochondrial DNA (mtDNA) in eukaryotic organisms occurs because of the selective destruction of paternal mtDNA molecules that may be present in the zygote. The elimination of sperm mtDNA is less efficient in interspecific crosses, and biparental inheritance of mtDNA has been observed in a variety of species. Because interspecific crosses are likely to be extremely rare in nature, parental inheritance of mtDNA has been deemed of little relevance to population genetics. The mtDNA of the parasitic trematode Schistosoma mansoni was examined for its utility in addressing epidemiological questions related to the transmission and spread of schistosomiasis. Prior to embarking on such experiments, we sought to confirm the mode of inheritance of this molecule using the highly polymorphic mtDNA minisatellite as a marker. In 3 separate crosses, mtDNA apparently identical to paternal DNA was observed in some individuals of the F2 and F3 generations. These observations thus suggest the intraspecific paternal inheritance of mtDNA across multiple generations in Schistosoma mansoni.

Susceptibility of Biomphalaria tenagophila and Biomphalaria straminea to Schistosoma mansoni infection detected by low stringency polymerase chain reaction.

In order to determine Schistosoma mansoni infection rates in Biomphalaria tenagophila and B. straminea, low stringency polymerase chain reaction (LS-PCR) technique was used as a complementary method to light exposure technique. LS-PCR has already been standardized in our laboratory to detect the trematode DNA in B. glabrata. Higher S. mansoni infection rates were detected using conventional method and LS-PCR. The parasite DNA profile was detected in both species after 7-day exposure to miracidia, using LS-PCR. This technique enables early detection of schistosomiasis transmission focuses, in endemic areas, before the beginning of cercariae shedding.

Mitotic histone H3 phosphorylation by the NIMA kinase in Aspergillus nidulans.

Phosphorylation of histone H3 serine 10 correlates with chromosome condensation and is required for normal chromosome segregation in Tetrahymena. This phosphorylation is dependent upon activation of the NIMA kinase in Aspergillus nidulans. NIMA expression also induces Ser-10 phosphorylation inappropriately in S phase-arrested cells and in the absence of NIMX(cdc2) activity. At mitosis, NIMA becomes enriched on chromatin and subsequently localizes to the mitotic spindle and spindle pole bodies. The chromatin-like localization of NIMA early in mitosis is tightly correlated with histone H3 phosphorylation. Finally, NIMA can phosphorylate histone H3 Ser-10 in vitro, suggesting that NIMA is a mitotic histone H3 kinase, perhaps helping to explain how NIMA promotes chromatin condensation in A. nidulans and when expressed in other eukaryotes.

Centrosomal and cytoplasmic Cdc2/cyclin B1 activation precedes nuclear mitotic events.

The activation of cdc2/cyclin B is the trigger for entry into mitosis. The mechanism of cdc2/cyclin B activation is complex, but the final step is the dephosphorylation of the Thr14 and Tyr15 residues on the cdc2 subunit, catalyzed by a member of the Cdc25 family of phosphatases. Cdc2/cyclin B1 accumulates at the centrosome in late G2 phase and has been implicated in the conversion of the centrosome from an interphase to a mitotic microtubule organizing center. Here we demonstrate biochemically that cdc2/cyclin B1 accumulates at the centrosome in late G2 as the inactive, phosphotyrosine 15 form and that the centrosomal cdc2/cyclin B1 can be activated in vitro by recombinant cdc25B. We provide evidence that a portion of the cdc2/cyclin B1 translocated into the nucleus in prophase is the inactive tyrosine-15-phosphorylated form. At this time the centrosomal and cytoplasmic cdc2/cyclin B1 is already active. This provides evidence that the activation of cdc2/cyclin B1 is initiated in the cytoplasm and that full activation of the translocated pool occurs in the nucleus.