Interaction between primary and secondary sporocysts of Schistosoma mansoni and the internal defence system of Biomphalaria resistant and susceptible to the parasite

The outcome of the interaction between Biomphalaria and Schistosoma mansoni depends on the response of the host internal defence system (IDS) and the escape mechanisms of the parasite. The aim of this study was to evaluate the responsiveness of the IDS (haemocytes and soluble haemolymph factors) of resistant and susceptible Biomphalaria tenagophila lineages and Biomphalaria glabrata lineages in the presence of in vitro-transformed primary sporocysts and secondary sporocysts obtained from infected B. glabrata. To do this, we assayed the cellular adhesion index (CAI), analysed viability/mortality, used fluorescent markers to evaluate the tegumental damage and transplanted secondary sporocysts. B. tenagophila Taim was more effective against primary and secondary sporocystes than the susceptible lineage and B. glabrata. Compared with secondary sporocysts exposed to B. tenagophila, primary sporocysts showed a higher CAI, a greater percentage of dead sporocysts and were labelled by lectin from Glycine max and Alexa-Fluor 488 fluorescent probes at a higher rate than the secondary sporocysts. However, the two B. tenagophila lineages showed no cercarial shedding after inoculation with secondary sporocysts. Our hypothesis that secondary sporocysts can escape the B. tenagophila IDS cannot be confirmed by the transplantation experiments. These data suggest that there are additional mechanisms involved in the lower susceptibilty of B. tenagophila to S. mansoni infection.

Standardisation of Western blotting to detect HTLV-1 antibodies synthesised in the central nervous system of HAM/TSP patients

Intrathecal synthesis of human T-lymphotropic virus type 1 (HTLV-1) antibodies (Abs) represents conclusive evidence of a specific immune response in the central nervous system of HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients. Western blotting (WB) for HTLV Abs in serum is a confirmatory test for HTLV-1 infection. The aim of this study was to standardise the Western blot to demonstrate the intrathecal pattern of Abs against HTLV-1 proteins in HAM/TSP patients. Paired cerebrospinal fluid (CSF) and serum samples were selected from 20 patients with definite HAM/TSP, 19 HTLV-1 seronegative patients and two HTLV-1 patients without definite HAM/TSP. The presence of reactive bands of greater intensity in the CSF compared to serum (or bands in only the CSF) indicated the intrathecal synthesis of anti-HTLV-1 Abs. All definite HAM/TSP patients presented with an intrathecal synthesis of anti-HTLV-1 Abs; these Abs were not detected in the control patients. The most frequent intrathecal targets of anti-HTLV-1 Abs were GD21, rgp46-I and p24 and, to a lesser extent, p19, p26, p28, p32, p36, p53 gp21 and gp46. The intrathecal immune response against env (GD21 and rgp46-I) and gag (p24) proteins represents the most important humoral pattern in HAM/TSP. This response may be used as a diagnostic marker, considering the frequent association of intrathecal anti-HTLV-1 Ab synthesis with HAM/TSP and the pathogenesis of this neurological disease.

Construction and characterisation of a complete reverse genetics system of dengue virus type 3

Dengue virulence and fitness are important factors that determine disease outcome. However, dengue virus (DENV) molecular biology and pathogenesis are not completely elucidated. New insights on those mechanisms have been facilitated by the development of reverse genetic systems in the past decades. Unfortunately, instability of flavivirus genomes cloned in Escherichia coli has been a major problem in these systems. Here, we describe the development of a complete reverse genetics system, based on the construction of an infectious clone and replicon for a low passage DENV-3 genotype III of a clinical isolate. Both constructs were assembled into a newly designed yeast- E. coli shuttle vector by homologous recombination technique and propagated in yeast to prevent any possible genome instability in E. coli . RNA transcripts derived from the infectious clone are infectious upon transfection into BHK-21 cells even after repeated passages of the plasmid in yeast. Transcript-derived DENV-3 exhibited growth kinetics, focus formation size comparable to original DENV-3 in mosquito C6/36 cell culture. In vitro characterisation of DENV-3 replicon confirmed its identity and ability to replicate transiently in BHK-21 cells. The reverse genetics system reported here is a valuable tool that will facilitate further molecular studies in DENV replication, virus attenuation and pathogenesis.

Governmental surveillance system of healthcare-associated infection in Brazil


Objective: This study aimed to describe the structure of governmental surveillance systems for Healthcare Associated Infection (HAI) in the Brazilian Southeastern and Southern States. Method: A cross-sectional, descriptive and exploratory study, with data collection by means of two-phases: characterization of the healthcare structure and of the HAI surveillance system. Results: The governmental teams for prevention and control of HAI in each State ranged from one to six members, having at least one nurse. All States implemented their own surveillance system. The information systems were classified into chain (n=2), circle (n=4) or wheel (n=1). Conclusion: Were identified differences in the structure and information flow from governmental surveillance systems, possibly limiting a nationwide standardization. The present study points to the need for establishing minimum requirements in public policies, in order to guide the development of HAI surveillance systems.


Characterization of the soil fertility and root system of restinga forests

The Restinga vegetation consists of a mosaic of plant communities, which are defined by the characteristics of the substrates, resulting from the type and age of the depositional processes. This mosaic complex of vegetation types comprises restinga forest in advanced (high restinga) and medium regeneration stages (low restinga), each with particular differentiating vegetation characteristics. The climate along the coast is tropical (Köppen). Of all ecosystems of the Atlantic Forest, Restinga is the most fragile and susceptible to anthropic disturbances. Plants respond to soil characteristics with physiological and morphological modifications, resulting in changes in the architecture (spatial configuration) of the root system. The purpose of this study was to characterize the soil fertility of high and low restinga forests, by chemical and physical parameters, and its relation to the root system distribution in the soil profile. Four locations were studied: (1) Ilha Anchieta State Park, Ubatuba; (2) two Ecological Stations of Jureia-Itatins and of Chauás, in the municipality of Iguape; (3) Vila de Pedrinhas in the municipality of Ilha Comprida; and (4) Ilha do Cardoso State Park, Cananeia. The soil fertility (chemical and physical properties) was analyzed in the layers 0-5, 0-10, 0-20, 20-40 and 40-60 cm. In addition, the distribution of the root system in the soil profile was evaluated, using digital images and the Spring program. It was concluded that the root system of all vegetation types studied is restricted to the surface layers, 0-10 and 10-20 cm, but occupies mainly the 0-10 cm layer (70 %); that soil fertility is low in all environments studied, with base saturation values below 16 %, since most exchange sites are occupied by aluminum; and that restinga vegetation is edaphic.

Nitrous oxide emissions from an intermittent aeration activated sludge system of an urban wastewater treatment plant

This study investigated the emission of N2O during the sequential aerated (60-min) and non-aerated (30-min) stages of an intermittent aeration cycle in an activated sludge wastewater treatment plant (WWTP). N2O emission occurred during both stages; however, emission was much higher during aeration. Air stripping is the major factor controlling transfer of N2O from the sewage to the atmosphere. The N2O emissions exclusively from the aeration tank represented 0.10% of the influent total nitrogen load and the per capita emission factor was almost 3 times higher than that suggested by the IPCC for inventories of N2O emission from WWTPs.

THE LOGICAL SYSTEM OF FREGE'S GRUNDGESTZE: A RATIONAL RECONSTRUCTION

This paper aims at clarifying the nature of Frege's system of logic, as presented in the first volume of the Grundgesetze . We undertake a rational reconstruction of this system, by distinguishing its propositional and predicate fragments. This allows us to emphasise the differences and similarities between this system and a modern system of classical second-order logic.

Effects of global climate changes on geographical distribution patterns of economically important plant species in cerrado

Different climate models, modeling methods and carbon emission scenarios were used in this paper to evaluate the effects of future climate changes on geographical distribution of species of economic and cultural importance across the Cerrado biome. As the results of several studies have shown, there are still many uncertainties associated with these projections, although bioclimatic models are still widely used and effective method to evaluate the consequences for biodiversity of these climate changes. In this article, it was found that 90% of these uncertainties are related to methods of modeling, although, regardless of the uncertainties, the results revealed that the studied species will reduce about 78% of their geographic distribution in Cerrado. For an effective work on the conservation of these species, many studies still need to be carried out, although it is already possible to observe that climate change will have a strong influence on the pattern of distribution of these species.

Annual evolution of global, direct and diffuse radiation and fractions in tilted surfaces

It was evaluated the annual evolution of global, direct and diffuse components of incident solar radiation on tilted surfaces to 12.85, 22.85 and 32.85º, facing north, in Botucatu, state of São Paulo, Brazil. The radiometric fractions were obtained for each component of the radiation in the aforementioned surfaces, through the ratio with the global and top of the atmosphere radiations. Seasonality was evaluated based on monthly averages of daily values. The measures occurred between 04/1998 and 07/2001 at 22.85º; 08/2001 and 02/2003 at 12.85º; and from 03/2003 to 12/2007 at 32.85º, with concomitant measures in the horizontal surface (reference). The levels of global and direct radiation on tilted surfaces were lower in summer and higher in the equinoxes when compared with the horizontal. The diffuse radiation on tilted surfaces was lower in most months, with losses of up to 65%. A trend of increasing differences occurred between horizontal and tilted surfaces with the increase of the angle in all the components and fractions of incident radiation. The annual evolution of rainfall and cloud cover ratio directly affected the atmospheric transmissivity of direct and diffuse components in the region.

Using an aerial system of remote sensing to detect different nutritional status in Brachiaria decumbens

The aim of this study was to use digital images acquired by cameras attached to a helium balloon to detect variation of the nutritional status in Brachiaria decumbens. The treatments consisted of five doses of nitrogen (0, 50, 100, 150 e 200kg ha-1) with six replications each, evaluated in a completely randomized statistical design. A remote sensing system composed of digital cameras and microcomputers was used for image acquisition, and a helium balloon lifted the cameras to the heights of 15, 20, 25 and 30m. A portable chlorophyll meter and analyses of leaf nitrogen content were used to make comparisons with data obtained by the remote sensing system. Data was acquired in two phases, in different climatic conditions. At the end of each phase, dry matter production was measured. Three vegetation indices were used to evaluate the detection of different nutritional status. The three indices were able to detect the effects of N doses. The indices constructed with the Green spectral band showed to be more efficient.

Effects of laparoscopy on the cardiorespiratory system of brown brocket deer (Mazama gouazoubira) anesthetized with ketamine/ xylazine/ midazolam combination and isoflurane

Laparoscopy is not widely used as a tool to perform assisted reproduction techniques in South American cervids; thus, scarce information in literature is available regarding its effects and appropriate anesthetic protocols to perform it. This study evaluated the effect of laparoscopy on heart rate (HR), respiration rate (RR), saturation of oxyhemoglobin (SpO2) and rectal temperature (RT) of six female brown brocket deer (Mazama gouazoubira) anesthetized with ketamine (5mg/kg), xylazine (0.3mg/kg), midazolam (0.5mg/kg) combination i.v. and isoflurane. Twelve laparoscopies were performed and each animal was used twice with a 40-day interval. After anesthetized, the animals were placed in dorsal recumbency to perform laparoscopy procedure using abdominal CO2 insufflations (14.2 ± 2.39mmHg; M ± SE). The main events of the laparoscopy procedure were divided into three periods: animal without (P1) and with abdominal insufflation (P2) and abdominal insufflation with the hips raised at 45º (P3). As a control, the animals were anesthetized again 40 days after the last laparoscopy, and were maintained in a dorsal recumbency for the same average duration of the previous anesthesia and no laparoscopy procedure was conducted. The period of anesthesia for the controls was also divided into P1, P2, and P3 considering the average duration of these periods in previous laparoscopies performed. Data were analyzed through the (ANOVA) variance analysis followed by Tukey test and values at P<0.05 were considered significant. No significant differences were observed in the parameters evaluated at P1, P2 and P3 between the animals submitted to laparoscopy and control. However, the RR mean between P1 (38.8 ± 4.42) and P3 (32.7 ± 4.81); and the RT mean between the P1 (38.2ºC ± 0.17), P2 (37.6ºC ± 0.19) and P3 (37.0ºC ± 0.21) varied significantly, independent of the laparoscopy. These data indicated that laparoscopy didn't cause any significant alterations in the cardiorespiratory parameters evaluated, even though the anesthetic protocol used can cause a reduction in the RT contributing to development of hypothermia during anesthesia.

Histopathological, immunohistochemical, and molecular study of BHV-5 infection in the central nervous system of experimentally infected calves

Bovine meningoencephalitis caused by BHV-5, a double-stranded DNA enveloped virus that belongs to the family Herpesviridae and subfamily Alphaherpesvirinae, is an important differential diagnosis of central nervous diseases. The aim of this study was to describe the histological changes in the central nervous system of calves experimentally infected with BHV-5 and compare these changes with the PCR and IHC results. Formalin-fixed paraffin-embedded central nervous system samples from calves previously inoculated with BHV-5 were microscopically evaluated and tested using IHC and PCR. All the animals presented with nonsuppurative meningoencephalitis. From 18 evaluated areas of each calf, 32.41% and 35.19% were positive by IHC and PCR, respectively. The telencephalon presented more accentuated lesions and positive areas in the PCR than other encephalic areas and was the best sampling area for diagnostic purposes. Positive areas in the IHC and PCR were more injured than IHC and PCR negative areas. The animal with neurological signs showed more PCR- and IHC-positive areas than the other animals.

Glial cells of the central nervous system of Bothrops jararaca (Reptilia, Ofidae): an ultrastructural study

Abstract Although ultrastructural characteristics of mature neuroglia in the central nervous system (CNS) are very well described in mammals, much less is known in reptiles, especially serpents. In this context, two specimens of Bothrops jararaca were euthanized for morphological analysis of CNS glial cells. Samples from telencephalon, mesencephalon and spinal cord were collected and processed for light and transmission electron microscopy investigation. Astrocytes, oligodendrocytes, microglial cells and ependymal cells, as well as myelin sheaths, presented similar ultrastructural features to those already observed in mammals and tended to maintain their general aspect all over the distinct CNS regions observed. Morphological similarities between reptilian and mammalian glia are probably linked to their evolutionary conservation throughout vertebrate phylogeny.

Floral biology and breeding system of three Ipomoea weeds

The floral biology of three weeds, Ipomoea cairica, I. grandifolia and I. nil (Convolvulaceae), was studied in Botucatu and Jaboticabal, São Paulo, in southeastern Brazil. The three species are melittophilous, with a varied set of floral visitors, but with some overlapping. Cluster analysis using Jacquard similarity index indicated a greater similarity among different plant species in the same locality than among the populations at different places, in relation to floral visitor sets. The promiscuous and opportunistic features of the flowers were shown, with such type of adaptation to pollination being advantageous to weeds since pollinator availability is unpredictable at ruderal environments.