Alternative Method for the Mass Rearing of Lutzomyia (Lutzomyia) cruzi (Diptera: Psychodidae) in a Laboratory Setting.

The understanding of the transmission dynamics of Leishmania spp. Ross as well as the epidemiology and spread of leishmaniasis is related to parasite-vector-host interactions. These interactions can be studied using specimens of a sand fly population reared in the laboratory, exposing individuals to experimental infection for the investigation of vector competence and parameters of the vectorial capacity of the species. The present study sought to describe an alternative method for the implantation of a Lutzomyia (Lutzomyia) cruzi colony with wild specimens captured in the municipality of Corumbá, Brazil. With Method 1, engorged females were individualized for oviposition. The eggs were transferred to an acrylic petri dish with a layer of plaster on the bottom, on which food was placed after hatching of the first larvae. With Method 2, females were kept in groups for oviposition in containers, in which soil and food were placed on their bottom for the larvae. In addition, the exposure time of the larvae to light was reduced in comparison with Method 1. With Method 2, a significantly greater number of specimens of Lu. cruzi was obtained. The ratio between the number of emerged adults and the females followed for oviposition was 0.42 with Method 1 and 2.75 with Method 2. The optimization of the rearing conditions for Lu. cruzi will enable the establishment of a colony providing a sufficient number of specimens to develop experimental infection by Leishmania as well as vectorial competence and some parameters of the vectorial capacity of this sand fly.

Ectoparasites and endoparasites of fish form networks with different structures.

Hosts and parasites interact with each other in a variety of ways, and this diversity of interactions is reflected in the networks they form. To test for differences in interaction patterns of ecto- and endoparasites we analysed subnetworks formed by each kind of parasites and their host fish species in fish-parasite networks for 22 localities. We assessed the proportion of parasite species per host species, the relationship between parasite fauna composition and host taxonomy, connectance, nestedness and modularity of each subnetwork (n = 44). Furthermore, we evaluated the similarity in host species composition among modules in ecto- and endoparasite subnetworks. We found several differences between subnetworks of fish ecto- and endoparasites. The association with a higher number of host species observed among endoparasites resulted in higher connectance and nestedness, and lower values of modularity in their subnetworks than in those of ectoparasites. Taxonomically related host species tended to share ecto- or endoparasites with the same interaction intensity, but the species composition of hosts tended to differ between modules formed by ecto- and endoparasites. Our results suggest that different evolutionary and ecological processes are responsible for organizing the networks formed by ecto- and endoparasites and fish.

Do educational methods affect students' ability to remove artificial carious dentine? A randomised controlled trial.

This trial aimed to evaluate the influence of two educational methods on students' ability to remove artificial carious dentine. Traditional lecture and lecture plus a live demonstration of artificial carious tissue removal were compared in a blind two-parallel-group design. Twenty-six students were randomly divided into two groups, and their skills were evaluated according to the following criteria: time spent on the dentine excavation procedure (in min), students' perceived confidence in conducting the procedure (graded assessed on a scale from 0 to 10), and the outcome of artificial carious tissue removal, evaluated by measuring the residual dyed artificial carious dentine layer (in µm). Statistical analyses were carried out using a t-test to compare the students' confidence and time spent on the procedure, and a two-way ANOVA was used to compare residual artificial decayed dentine with educational methods and tooth region (incisal, medium, and cervical thirds) as factors. There were no differences between the methods regarding excavation time (P = 0.898) and students' confidence (P = 0.382). The residual artificial carious dentine results showed that the educational method (P < 0.001) and cavity region (P < 0.001) were statistically significant, as was their interaction (P = 0.040). The lecture plus live demonstration group presented the best results for artificial caries removal. Although there were no differences between the two groups for the cervical region, the best results for the lecture plus live demonstration group was in the other two-thirds of the tooth.

Lipid metabolism of commercial layers fed diets containing aflatoxin, fumonisin, and a binder.

Aflatoxins (AF) and fumonisins (FU) are a major problem faced by poultry farmers, leading to huge economic losses. This experiment was conducted to determine the effects of AF (1 mg/kg of feed) and FU (25 mg/kg of feed), singly or in combination, on the lipid metabolism in commercial layers and investigate the efficacy of a commercial binder (2 kg/t of feed) on reducing the toxic effects of these mycotoxins. A total of 168 Hisex Brown layer hens, 37 wk of age, were randomized into a 3 × 2 + 1 factorial arrangement (3 diets with no binder containing AF, FU, and AF+FU; 3 diets with binder containing AF, FU, and AF+FU; and a control diet with no mycotoxins and binders), totaling 7 treatments. The hens contaminated with AF showed the characteristic effects of aflatoxicosis, such as a yellow liver, resulting from the accumulation of liver fat, lower values of plasma very low-density lipoprotein and triglycerides, and higher relative weight of the kidneys and liver. Hepatotoxic and nephrotoxic effects of FU were not observed in this study. On the other hand, the FU caused a reduction in small intestine length and an increase in abdominal fat deposition. The glucan-based binder prevented some of the deleterious effects of these mycotoxins, particularly the effects of AF on hepatic lipid metabolism, kidney relative weight, and FU in the small intestine.

Behavioral aspects of Lutzomyia longipalpis (Diptera: Psychodidae) in urban area endemic for visceral leishmaniasis.

The study of some of the behavioral aspects of the main vector of Leishmania infantum chagasi Cunha & Chagas in the Americas, Lutzomyia longipalpis (Lutz & Neiva), such as dispersion, population size, and vector survival rates, is important for the elucidation of the mechanisms of visceral leishmaniasis transmission. These parameters were studied by means of capture-mark-release-recapture experiments in an urban area of Campo Grande municipality, an endemic area of visceral leishmaniasis, situated in Mato Grosso do Sul state, Brazil. Six capture-mark-release-recapture experiments were undertaken between November 2009 and November 2010 and once in January 2012 with a view to assessing the population size and survival rate of Lu. longipalpis. The insects were released in a peridomicile surrounded by 13 residences. The recaptures were undertaken with automatic light traps for four consecutive weeks after release in the surrounding area. In total, 3,354 sand flies were captured, marked, and released. The overall recapture rate during the capture-mark-release-recapture experiments was 4.23%, of which 92.45% were recaptured at the release site, indicating limited dispersal. The greatest distance recorded from the release site was 165 m for males and 241 m for females. The male daily survival rate, calculated on the basis of regressions from the numbers of marked recaptured insects during the 15 successive days after release was 0.897. The estimated male population size measured by the Lincoln Index was 10,947.127. Though Lu. longipalpis presented a limited dispersion the physical barriers typical of urban environments did not prevent the sand flies from flying long distances.

Conventional caries removal and sealed caries in permanent teeth: a microbiological evaluation.

OBJECTIVES: The aim of this study was to compare microbiological infection after conventional carious dentine removal with incomplete carious dentine removal and sealing. METHODS: Eighty-seven patients (12-50 years of age) under treatment at the Dental Clinics of the Federal University of Rio Grande do Sul (UFRGS), Brazil, participated in the study. The patients presented 90 posterior permanent teeth with primary caries. The lesions were coronal, active, and reached at least the middle third of the dentine. None of the teeth exhibited spontaneous pain, sensitivity to percussion or apical pathology (detected through radiographic exams). Pulp sensibility was confirmed by the cold test. The lesions were divided into 2 experimental groups: complete caries removal (CCR) based on hardness criteria (n=60 lesions) and incomplete caries removal (ICR) and sealing (n=32 lesions). Microbiological samples were obtained from the initial demineralized dentine, after CCR and after ICR-Seal. RESULTS: The number of anaerobic and aerobic bacteria, lactobacilli, and mutans streptococci decreased at the end of treatment (p<0.05). Significantly less anaerobic bacteria (p<0.01), aerobic bacteria (p=0.02), and mutans streptococci (p<0.01) growth was observed after ICR-Seal compared to CCR. The difference in lactobacilli was insignificant (p=0.08). The amount of bacteria detected after conventional caries removal was higher than that which remained in sealed caries lesions. CONCLUSIONS: The results suggest it is not necessary to remove all carious dentine before the restoration is placed because over time, sealing of carious dentine results in lower levels of infection than traditional dentine caries removal. CLINICAL SIGNIFICANCE: The results of this study indicate that sealed carious dentine was less infected than the remaining dentine left after conventional caries removal and sealing. Our results support treatment of deep carious lesions in one session with incomplete removal of carious dentine.

Trophic ecomorphology of Siluriformes (Pisces, Osteichthyes) from a tropical stream.

The present study analysed the relationship between morphology and trophic structure of Siluriformes (Pisces, Osteichthyes) from the Caracu Stream (22º 45' S and 53º 15' W), a tributary of the Paraná River (Brazil). Sampling was carried out at three sites using electrofishing, and two species of Loricariidae and four of Heptapteridae were obtained. A cluster analysis revealed the presence of three trophic guilds (detritivores, insectivores and omnivores). Principal components analysis demonstrated the segregation of two ecomorphotypes: at one extreme there were the detritivores (Loricariidae) with morphological structures that are fundamental in allowing them to fix themselves to substrates characterised by rushing torrents, thus permitting them to graze on the detritus and organic materials encrusted on the substrate; at the other extreme of the gradient there were the insectivores and omnivores (Heptapteridae), with morphological characteristics that promote superior performance in the exploitation of structurally complex habitats with low current velocity, colonised by insects and plants. Canonical discriminant analysis revealed an ecomorphological divergence between insectivores, which have morphological structures that permit them to capture prey in small spaces among rocks, and omnivores, which have a more compressed body and tend to explore food items deposited in marginal backwater zones. Mantel tests showed that trophic structure was significantly related to the body shape of a species, independently of the phylogenetic history, indicating that, in this case, there was an ecomorphotype for each trophic guild. Therefore, the present study demonstrated that the Siluriformes of the Caracu Stream were ecomorphologically structured and that morphology can be applied as an additional tool in predicting the trophic structure of this group.

Multiple opioid receptors mediate the hypotensive response induced by central 5-HT(3) receptor stimulation.

The aim of the present work was to investigate the role of brain µ, κ and δ opioid receptors in the central serotonergic mechanisms regulating blood pressure in rats. The data obtained show that: (1) pharmacological activation of central 5-HT(3) receptors yields a significant decrease in blood pressure; (2) the blockade of those receptors by a selective antagonist induces an acute hypertensive response; (3) the pharmacological blockade of central opioid receptors by three different opioid antagonists exhibiting variable degrees of selectivity to µ, κ and δ opioid receptors always suppressed the hypotensive response induced by central 5-HT(3) receptor stimulation; (4) the blockade of opioid receptors by the same opioid antagonists that impaired the hypotensive effect of central 5-HT(3) receptor stimulation failed to modify blood pressure in animals not submitted to pharmacological manipulations of central 5-HT(3) receptor function. It is shown that a 5-HT(3) receptor-dependent mechanism seems to be part of the brain serotonergic system that contributes to cardiovascular regulation since the hypertensive response observed after ondansetron administration indicates that central 5-HT(3) receptors exert a tonic inhibitory drive on blood pressure. Furthermore, the data obtained here clearly indicate that the hypotensive response observed after pharmacological stimulation of central 5-HT(3) receptors depends on the functional integrity of brain µ, κ and δ opioid receptors, suggesting that a functional interaction between serotonergic and opiatergic pathways in the brain is part of the complex, multifactorial system that regulates blood pressure in the central nervous system.

Searching for bioindicators of forest fragmentation: passerine birds in the Atlantic forest of southeastern Brazil.

Aiming to evaluate the potential of Passerine birds as bioindicators of forest fragmentation, we studied the avifauna in the mountain region of the state of Rio de Janeiro by mist-netting between 2001 and 2005. We sampled six sites, including four small fragments (from 4 to 64 ha) in an agricultural area (Teresópolis), one second-growth forest (440 ha - Miguel Pereira) and a continuous forest (10,600 ha, Serra dos Orgãos National Park - SONP). Indicator Species analysis and a Monte Carlo test were run to detect associations between species and sites, considering at least 30% of perfect indication and a significant value for the statistical test, only considering species with at least 10 captures. A total of 30 Passerine birds were sampled, and due to their association to the largest area (SONP), we considered Sclerurus scansor, Mionectes rufiventris, Chiroxiphia caudata and Habia rubica as the best indicators for this area. Five species were more captured not by chance in the 440 ha second-growth: Conopophaga melanops, Myiobius barbatus, Myrmeciza loricata, Philydor atricapillus and Schiffornis virescens and no species were related to any small fragment. This analysis has identified specialized Passerine species in many aspects (foraging substract, needs for nesting, rarity status), and it can be a valuable tool for detecting possible bioindicators.

Searching for bioindicators of forest fragmentation: passerine birds in the Atlantic forest of southeastern Brazil / Busca por bioindicadores de fragmentação florestal: aves Passeriformes na Floresta Atlântica do Sudeste do Brasil

Aiming to evaluate the potential of Passerine birds as bioindicators of forest fragmentation, we studied the avifauna in the mountain region of the state of Rio de Janeiro by mist-netting between 2001 and 2005. We sampled six sites, including four small fragments (from 4 to 64 ha) in an agricultural area (Teresópolis), one second-growth forest (440 ha - Miguel Pereira) and a continuous forest (10,600 ha, Serra dos Órgãos National Park - SONP). Indicator Species analysis and a Monte Carlo test were run to detect associations between species and sites, considering at least 30 percent of perfect indication and a significant value for the statistical test, only considering species with at least 10 captures. A total of 30 Passerine birds were sampled, and due to their association to the largest area (SONP), we considered Sclerurus scansor, Mionectes rufiventris, Chiroxiphia caudata and Habia rubica as the best indicators for this area. Five species were more captured not by chance in the 440 ha second-growth: Conopophaga melanops, Myiobius barbatus, Myrmeciza loricata, Philydor atricapillus and Schiffornis virescens and no species were related to any small fragment. This analysis has identified specialized Passerine species in many aspects (foraging substract, needs for nesting, rarity status), and it can be a valuable tool for detecting possible bioindicators.

Para avaliar o potencial de Passeriformes como bioindicadores de fragmentação florestal na Mata Atlântica, estudamos a avifauna da região serrana do Rio de Janeiro, utilizando capturas com redes de neblina entre 2001 e 2005. Foram amostradas seis áreas, sendo quatro pequenos fragmentos (de 4 a 64 ha) em áreas agrícolas (Teresópolis), uma mata secundária com cerca de 40 anos (440 ha - Miguel Pereira) e uma área de mata contínua de 10600 hectares (Parque Nacional da Serra dos Órgãos - PARNASO). A análise de espécies indicadoras e o teste de Monte Carlo foram empregados para que se verificassem as espécies mais associadas a cada local amostrado, considerando-se um mínimo de 30 por cento de indicação e valor significativo para o teste mencionado, utilizando-se somente espécies com 10 ou mais capturas. Foi amostrado um total de 30 espécies e, pela sua associação com a maior área (PARNASO), consideramos Sclerurus scansor, Mionectes rufiventris, Chiroxiphia caudata e Habia rubica como as melhores indicadoras. Cinco espécies foram mais capturadas não ao acaso na área de 440 ha: Conopophaga melanops, Myiobius barbatus, Myrmeciza loricata, Philydor atricapillus e Schiffornis virescens. Nenhuma espécie foi associada aos pequenos fragmentos. Como esta análise identificou espécies de Passeriformes especialistas em diversos aspectos (substrato de forrageio, locais de nidificação, status de raridade), ela pode ser uma ferramenta útil na detecção de possíveis bioindicadores.

Deep caries lesions after incomplete dentine caries removal: 40-month follow-up study.

Radiographic changes after indirect pulp capping (32 teeth in 27 patients) were studied for up to 36-45 months. Radiolucent zone (RZ) depth and tertiary dentine formation were assessed qualitatively and changes in radiographic density (by image subtraction) in RZ and control areas (CA) were estimated. During follow-up there were 1 pulp necrosis, 1 pulp exposure, 3 fractures and 3 withdrawals. Twelve cases showed decreased RZ depth and 4 displayed tertiary dentine. No changes with time in density of CA or RZ, or in the difference between them, were observed. It is concluded that indirect pulp capping arrests lesion progression, suggesting that complete dentine caries removal is not essential for caries control.

The monitoring of deep caries lesions after incomplete dentine caries removal: results after 14-18 months.

This paper aims to assess radiographic changes after incomplete carious dentine removal and tooth sealing. Thirty-two teeth with deep caries lesions were studied. The treatment consisted of incomplete excavation, application of a Ca(OH)2 layer, sealing temporarily for a 6- to 7-month period and then restoration. Standardised bitewing radiographs were taken immediately after the temporary sealing and at 6- to 7- and 14- to 18-month intervals. The digitised images were analysed blind by image subtraction. The quantitative analyses subtractions were performed in the radiolucent zone (RZ) beneath the restoration and in two adjacent control areas (CA). Two cases were lost during the 6- to 7-month period (one pulp necrosis and one pulp exposure during removal of the provisional sealing). No difference (p > 0.05) was observed in the radiographic density of the CA and the RZ in the two experimental periods. The mean and standard deviation (grey tonalities scale) were 129.42+/-5.83 and 127.65+/-4.67 (control areas) and 132.96+/-7.41 and 132.90+/-5.99 (RZ) for the first and second experimental periods, respectively. The radiographic density of the CA differed from the RZ (Tukey test, p < 0.001). Interference in environmental conditions by partial dentine caries removal and tooth sealing arrests lesion progression, suggesting that complete dentine caries removal is not essential to control caries progression.

Fish diversity along spatial gradients in the Itaipu Reservoir, Paraná, Brazil.

Local and turnover patterns of fish diversity in the Itaipu Reservoir were evaluated and related to a longitudinal or river-dam gradient (composed of riverine, transitional, and lacustrine zones) and to transversal or upstream-downstream gradients of the tributaries (composed of lotic and lentic stretches of tributaries and reservoir shores). Thirteen stations were sampled quarterly during 2 years. A total of 85 fish species were caught. Local (alpha) and turnover (beta) patterns of fish diversity showed significant differences in reservoir spatial gradients. Along the longitudinal gradient, total and alpha-diversity were the highest in the riverine and transitional zones of the reservoir and lowest in the lacustrine zone. Along the transversal gradient, total and alpha-diversity increased from the lotic stretches of the tributaries to the reservoir shores. The lotic and lentic stretches of the tributaries presented the highest beta-diversity values, indicating heterogeneity in species compositions among the sub-basins. We conclude with recommendations for reservoir management based on the results of this study.

Fish diversity along spatial gradients in the Itaipu Reservoir, Paraná, Brazil

Padrões locais e regionais de diversidade de peixes no reservatório de Itaipu foram avaliados e relacionados aos gradientes longitudinal ou rioûrepresa (dividido nas zonas fluvial, intermediária e lacustre) e transversal ou cabeceiraûfoz dos tributários (dividido em trechos lóticos e lênticos dos tributários e margens do reservatório). As coletas foram realizadas trimestralmente durante dois anos. Foram capturadas 85 espécies. Os padrões de diversidade local (a) e turnover (b) apresentaram diferenças significativas ao longo dos gradientes espaciais. Ao longo do gradiente longitudinal, as diversidades total e a foram maiores nas zonas fluvial e intermediária. Ao longo do gradiente transversal, as diversidades total e a aumentaram gradualmente do trecho lótico dos tributários às margens do reservatório. Os trechos lóticos e lênticos dos tributários apresentaram os maiores valores de diversidade b, indicando a heterogeneidade na composição de espécies das sub-bacias. Com base nos resultados deste estudo, recomendamos algumas ações de manejo para o reservatório.

The use of a muscle graft to repair a segmentary nerve defect. An experimental study using the sciatic nerve of rats as model.

The use of a devitalized skeletal muscle graft and conventional nerve graft to repair a 5mm long segmentary sciatic nerve lesion was studied in rats by means of functional, morphometric and spinal cord motor neuron cell response evaluation. Thirty-four rats were used and divided into four groups according to the procedure: (1) sham operation; (2) conventional nerve grafting; (3) muscle grafting; (4) unrepaired lesion. The sciatic functional index (SFI) was evaluated every fortnight up to the 105th postoperative day by measuring three parameters in the rats' footprint. The animals of Groups 2 and 3 presented initial complete functional loss, followed by slow but steady recovery, with final similar SFIs. The histologic and morphometric studies showed an increased small diameter/thin myelin sheath nerve fiber density distally to the lesion site for both types of graft. An increased population of motor neurons was observed in the anterior horn of the lumbar spinal cord segment with both types of grafts, but not in the control groups. The SFI, histologic and morphometric data did not differ significantly between the two types of graft, thus indicating a similar behavior. The authors conclude that a 5mm long skeletal muscle graft works as well as a conventional nerve graft.

Correlation between functional index and morphometry to evaluate recovery of the rat sciatic nerve following crush injury: experimental study.

An experimental study on the correlation between functional and morphologic recovery of crushed sciatic nerves was carried out in rats. The sciatic nerve of 33 rats, divided into three groups, was submitted to controlled crushing injury on a 5-mm long segment, in a universal testing machine for 10 min with three different loads (100, 500, and 15,000 g, respectively). Functional recovery was evaluated, using a modified sciatic functional index (SFI) at weekly intervals up to the 60th postoperative day, at which time, the animals were sacrificed for histologic and morphometric studies of the nerves. Results were compared with those of normal untouched nerves and nerves submitted to segmentary resection without repair. Initial loss of function was observed in all animals with crush injury, but recovery to a nearly normal SFI occurred after progressively longer intervals (25, 39, and 53 days), as a function of load. Nerve-fiber density was increased in the groups submitted to lower loads, but statistically significantly decreased in the animals submitted to the 15,000-g crush. The authors conclude that the SFI is directly correlated with nerve-fiber density and, therefore, is an adequate tool for evaluating sciatic functional deficiency in the rat.

[Physical examination in children: a nurse's guide]. / Exame físico na criança: um guia para o enfermeiro.

The physical exam is constituted inside in an important stage of the evaluation process to the child, through which the nurse enriches the information obtained during the interview and it bases the nursing care. At the present time, by means of the nurses growing interest in applying the nursing process in whole its stages, the physical exam has been occupying prominence place, for allowing the nurses to know the its customers' needs, in what says respect to its physical and physiologic aspects, allowing the identification of the nursing diagnoses, besides serving as resource for evaluation effective of the nursing interventions. This study, of bibliographical character, objectified to elaborate a guide for accomplishment of the physical exam in the child, to be used by nurses, during the evaluation process to the child. In him we presented the physical exam in a systematic way, using the sequence cephalic-flow, presenting the possible normal and abnormal discoveries to the we examine the child.

Isolation of an aspartic proteinase precursor from the egg of a hard tick, Boophilus microplus.

An aspartic proteinase precursor, herein named BYC (Boophilus Yolk pro-Cathepsin) was isolated from eggs of the hard tick, Boophilus microplus. As judged by electrophoresis on sodium dodecyl sulfate polyacrylamide slab gel (SDS-PAGE), purified BYC presented 2 bands of 54 and 49 kDa, bearing the same NH2-terminal amino acid sequence. By Western blot analysis, BYC was also found in the haemolymph, indicating an extraovarian site of synthesis. Several organs were incubated in culture medium with [35S]methionine, and only the gut and fat body showed synthesis of BYC polypeptides. Protein sequencing of both the NH2-terminal and an internal sequence obtained after cyanogen bromide (CNBr) cleavage of BYC revealed homology with several aspartic proteinase precursors. Incubation at pH 3.5 resulted in autoproteolysis of BYC, which produced the mature form of the enzyme, that displayed pepstatin-sensitive hydrolytic activity against haemoglobin. Western blot analysis using anti-BYC monoclonal antibodies showed proteolytic processing of BYC during embryogenesis and suggested activation of the enzyme during development. A role of BYC in degradation of vitellin, the major yolk protein of tick eggs, is discussed.

Sequence and phylogenetic analysis of glycoprotein 120 of an HIV type 1 variant (GWGR) prevalent in Brazil.

PIP: Findings are reported from a study conducted to fully sequence the gp120 gene from a Brazilian HIV-1 isolate containing the GWGR motif and compare it to the Brazilian B and F sequences already described. Genomic DNA isolated from six patients in an ongoing HIV cohort study was screened for the presence of the viral V3 loop GWGR motif. Sequence analysis revealed that BZ(GWGR)1 is closely related to the North American MN prototype strain, with 80.1% amino acid identity and 89.1% nucleic acid similarity, and with 6 deletions and 11 insertions. Large differences were, however, observed when the V1 and V2 regions of MN and BZ(GWGR)1 were compared. Tree analysis based upon amino acid sequences and the four Brazilian isolates introduced in the analysis indicate that BZ(GWGR)1 belongs to the HIV-1 B subtypes. Several features of BZ(GWGR)1 suggest that some biological advantage may be derived from the differences between that variant and the American/European prototype strain.^ieng