Quantitative method of viral pollution determination for large volume of water using ferric hydroxide gel impregnated on the surface of glassfibre cartridge

Quantitative method of viral pollution determination for large volume of water using ferric hydroxide gel impregnated on the surface of glassfibre cartridge filter. The use of ferric hydroxide gel, impregnated on the surface of glassfibre cartridge filter enable us to recover 62.5% of virus (Poliomylitis type I, Lsc strain) exsogeneously added to 400 liters of tap-water. The virus concentrator system consists of four cartridge filters, in which the three first one are clarifiers, where the contaminants are removed physically, without significant virus loss at this stage. The last cartridge filter is impregnated with ferric hydroxide gel, where the virus is adsorbed. After the required volume of water has been processed, the last filter is removed from the system and the viruses are recovered from the gel, using 1 liter of glycine/NaOH buffer, at pH 11. Immediately the eluate is clarified through series of cellulose acetate membranes mounted in a 142mm Millipore filter. For the second step of virus concentration, HC1 1N is added slowly to the eluate to achieve pH 3.5-4. MgC1, is added to give a final concentration of 0.05M and the viruses are readsorbed on a 0.45 , porosity (HA) cellulose acetate membrane, mounted in a 90 mm Millipore filter. The viruses are recovered using the same eluent plus 10% of fetal calf serum, to a final volume of 3 ml. In this way, it was possible to concentrate virus from 400 liters of tap-water, into 1 liter in the first stage of virus concentration and just to 3 ml of final volume in a second step. The efficiency, simplicity and low operational cost, provded by the method, make it feasible to study viral pollution of recreational and tap-water sources.

Theoretical estimates of consumable food and probability of acquiring food in larvae of Chrysomya putoria (Diptera: Calliphoridae)

An indirect estimate of consumable food and probability of acquiring food in a blowfly species, Chrysomya putoria, is presented. This alternative procedure combines three distinct models to estimate consumable food in the context of the exploitative competition experienced by immature individuals in blowfly populations. The relevant parameters are derived from data for pupal weight and survival and estimates of density-independent larval mortality in twenty different larval densities. As part of this procedure, the probability of acquiring food per unit of time and the time taken to exhaust the food supply are also calculated. The procedure employed here may be valuable for estimations in insects whose immature stages develop inside the food substrate, where it is difficult to partial out confounding effects such as separation of faeces. This procedure also has the advantage of taking into account the population dynamics of immatures living under crowded conditions, which are particularly characteristic of blowflies and other insects as well.

Quantitative phenetics and taxonomy of some phlebotomine taxa

Elucidating the evolution of Phlebotominae is important not only to revise their taxonomy, but also to help understand the origin of the genus Leishmania and its relationship with humans. Our study is a phenetic portrayal of this history based on the genetic relationships among some New Word and Old Word taxa. We used both multilocus enzyme electrophoresis and morphometry on 24 male specimens of the Old Word genus Phlebotomus (with three of its subgenera: Phlebotomus, Spelaeophlebotomus and Australophlebotomus), and on 67 male specimens of the three New World genera, Warileya, Brumptomyia and Lutzomyia, (with three subgenera of Lutzomyia: Lutzomyia, Oligodontomyia and Psychodopygus). Phenetic trees derived from both techniques were similar, but disclosed relationships that disagree with the present classification of sand flies. The need for a true evolutionary approach is stressed.

Quantitative morphological evidence for incipient species within Lutzomyia quinquefer (Diptera: Psychodidae)

Morphological variation among geographic populations of the New World sand fly Lutzomyia quinquefer (Diptera, Phlebotominae) was analyzed and patterns detected that are probably associated with species emergence. This was achieved by examining the relationships of size and shape components of morphological attributes, and their correlation with geographic parameters. Quantitative and qualitative morphological characters are described, showing in both sexes differences among local populations from four Departments of Bolivia. Four arguments are then developed to reject the hypothesis of environment as the unique source of morphological variation: (1) the persistence of differences after removing the allometric consequences of size variation, (2) the association of local metric properties with meristic and qualitative attributes, rather than with altitude, (3) the positive and significant correlation between metric and geographic distances, and (4) the absence of a significant correlation between altitude and general-size of the insects.

Phenotypic, functional, and quantitative characterization of canine peripheral blood monocyte-derived macrophages

The yield as well as phenotypic and functional parameters of canine peripheral blood monocyte-derived macrophages were analyzed. The cells that remained adherent to Teflon after 10 days of culture had high phagocytic activity when inoculated with Leishmania chagasi. Flow cytometric analysis demonstrated that more than 80% of cultured cells were positive for the monocyte/macrophage marker CD14.

Reliability of current estimates of schistosomiasis prevalence in the Rainforest Zone of the state of Pernambuco, Northeastern Brazil

The Program for Schistosomiasis Control within the Unified Health System (PCE-SUS) was implemented by 1999 in the Rainforest Zone or "Zona da Mata" of Pernambuco (ZMP) aiming to carry out biennial stool surveys of whole populations through municipal health organs followed by treatment of the positives through the local units of the Family Health Program (PSF). Yearly reports from the Health Department of Pernambuco State (SES/PE) from 2002 to 2004 on the PCE-SUS surveys were assessed to evaluate whether the current estimates of prevalence in the municipalities of the ZMP are based on reliable samples so as to allow considerations on the real situation of schistosomiasis in that area. The surveys carried out in that period did not follow the major principles underlying sampling design, thus posing problems in both precision and validity of the estimates. Only 12 out of 43 municipalities had minimally reliable estimates: five with moderate prevalence (10-50%) and seven with low prevalence (< 10%). Surveys with appropriate sampling procedures aimed either at representative target groups (school-aged children) or communities are recommended for the ZMP and other endemic areas not only to provide reliable information on the current situation of schistosomiasis but also to plan adequate control strategies.

Quantitative paleoparasitology applied to archaeological sediments

Three techniques to extract parasite remains from archaeological sediments were tested. The aim was to improve the sensibility of recommended paleoparasitological techniques applied in archaeological remains. Sediment collected from the pelvic girdle of a human body found in Cabo Vírgenes, Santa Cruz, Argentina, associated to a Spanish settlement founded in 1584 known as Nombre de Jesús, was used to search for parasites. Sediment close to the skull was used as control. The techniques recommended by Jones, Reinhard, and Dittmar and Teejen were used and compared with the modified technique presented here, developed to improve the sensibility to detect parasite remains. Positive results were obtained only with the modified technique, resulting in the finding of Trichuris trichiura eggs in the sediment.

Plasmid-based controls to detect rpoB mutations in Mycobacterium tuberculosis by quantitative polymerase chain reaction-high-resolution melting

Quantitative polymerase chain reaction-high-resolution melting (qPCR-HRM) analysis was used to screen for mutations related to drug resistance in Mycobacterium tuberculosis. We detected the C526T and C531T mutations in the rifampicin resistance-determining region (RRDR) of the rpoB gene with qPCR-HRM using plasmid-based controls. A segment of the RRDR region from M. tuberculosis H37Rv and from strains carrying C531T or C526T mutations in the rpoB were cloned into pGEM-T vector and these vectors were used as controls in the qPCR-HRM analysis of 54 M. tuberculosis strains. The results were confirmed by DNA sequencing and showed that recombinant plasmids can replace genomic DNA as controls in the qPCR-HRM assay. Plasmids can be handled outside of biosafety level 3 facilities, reducing the risk of contamination and the cost of the assay. Plasmids have a high stability, are normally maintained in Escherichia coli and can be extracted in large amounts.

Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation

Human herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance.

Biomass and abundance of termites in three remnant areas of Atlantic Forest in northeastern Brazil

Termite biomass and abundance were estimated in three remnant areas of Atlantic Forest in northeastern Brazil: Reserva Biológica de Pedra Talhada-RBPT (Alagoas State), Parque Estadual Dois Irmãos-PEDI (Pernambuco State), and Área de Proteção Ambiental Mata do Buraquinho-APMB (Paraíba State). A quantitative sampling of the termite assemblages present on trunks and leaf litter, in nests, and in the soil was undertaken during the months of June and July (rainy season) and November and December (dry season) of 2000. Thirty seven species of termites were encountered and quantified in the three remnant forest areas. The termite biomass varied from 10.09 to 11.30 g (fresh weight)/m² and abundance varied from 4911 to 5663 individuals/m². The subfamily Nasutitermitinae was dominant in terms of the number of species, biomass, and abundance. The soil-feeders were the most abundant at the three sites, while wood-feeders had the largest biomass at least at the PEDI and APMB sites. Among the microhabitats quantified, termites were most frequently encountered in the soil, being most heavily concentrated at depths of 0-20 cm. Two species stood out in terms of their biomass and abundance: Embiratermes parvirostris Constantino, on the soil, and Nasutitermes corniger (Motschulsky), on decomposing trunks and in nests. Estimates of biomass and abundance suggest that termites play an important role in the decomposition of the plant necromass in remnant forest areas of the Atlantic Forest in northeastern Brazil.

Nutritional ecology of blowflies (Diptera, Calliphoridae): estimates of critical larval weight for pupation on two different

Under natural environmental conditions, blowflies utilize discrete and ephemeral feeding resources such as decaying carcasses. Competition for food on such feeding substrates is usually very severe, and only the individuals that are capable of attaining the critical larval weight for pupation will be able to survive. This critical weight is hitherto unknown for several blowfly species; therefore, the current work is aimed at obtaining such a critical value for four blowfly species of the genera Chrysomya and Lucilia, deploying two types of feeding substrate, namely, artificial diet and macerated bovine meat. On the whole, the critical weights ranged from 30 to 35 mg. The lowest larval weight which permitted pupation was 30.0 mg for Chrysomya megacephala reared on macerated bovine meat. This species was also the best adapted to pupation at low larval weights in relation to the maximum larval weight for males. Regarding the pupation of females, the best-adapted individual was a C. albiceps specimen exhibiting a critical weight that was equal to 39.20 % of the maximum value obtained. Concerning all the species and diet types, the female individuals exhibited the lowest critical weights that produced viable pupae, probably representing an evolutionary strategy that favoured the survival of females, responsible for the egg formation, contributing to the establishment of future generations. Regarding the loss (in percentage) of adult biomass in relation to the third instar larvae, the females of C. megacephala lost less weight than males in both feeding substrates. On the other hand, such a loss of weight occurred in males of C. albiceps and L. cuprina.

The nitrogen sufficiency index underlying estimates of nitrogen fertilization requirements of common bean

Chlorophyll determination with a portable chlorophyll meter can indicate the period of highest N demand of plants and whether sidedressing is required or not. In this sense, defining the optimal timing of N application to common bean is fundamental to increase N use efficiency, increase yields and reduce the cost of fertilization. The objectives of this study were to evaluate the efficiency of N sufficiency index (NSI) calculated based on the relative chlorophyll index (RCI) in leaves, measured with a portable chlorophyll meter, as an indicator of time of N sidedressing fertilization and to verify which NSI (90 and 95 %) value is the most appropriate to indicate the moment of N fertilization of common bean cultivar Perola. The experiment was carried out in the rainy and dry growing seasons of the agricultural year 2009/10 on a dystroferric Red Nitosol, in Botucatu, São Paulo State, Brazil. The experiment was arranged in a randomized complete block design with five treatments, consisting of N managements (M1: 200 kg ha-1 N (40 kg at sowing + 80 kg 15 days after emergence (DAE) + 80 kg 30 DAE); M2: 100 kg ha-1 N (20 kg at sowing + 40 kg 15 DAE + 40 kg 30 DAE); M3: 20 kg ha-1 N at sowing + 30 kg ha-1 when chlorophyll meter readings indicated NSI < 95 %; M4: 20 kg ha-1 N at sowing + 30 kg ha-1 N when chlorophyll meter readings indicated NSI < 90 % and, M5: control (without N application)) and four replications. The variables RCI, aboveground dry matter, total leaf N concentration, production components, grain yield, relative yield, and N use efficiency were evaluated. The RCI correlated with leaf N concentrations. By monitoring the RCI with the chlorophyll meter, the period of N sidedressing of common bean could be defined, improving N use efficiency and avoiding unnecessary N supply to common bean. The NSI 90 % of the reference area was more efficient to define the moment of N sidedressing of common bean, to increase N use efficiency.

Quantitative Assessment of Soil Physical Quality in Northern China Based on S-theory

ABSTRACT Quantitative assessment of soil physical quality is of great importance for eco-environmental pollution and soil quality studies. In this paper, based on the S-theory, data from 16 collection sites in the Haihe River Basin in northern China were used, and the effects of soil particle size distribution and bulk density on three important indices of theS-theory were investigated on a regional scale. The relationships between unsaturated hydraulic conductivityKi at the inflection point and S values (S/hi) were also studied using two different types of fitting equations. The results showed that the polynomial equation was better than the linear equation for describing the relationships between -log Ki and -logS, and -log Kiand -log (S/hi)2; and clay content was the most important factor affecting the soil physical quality index (S). The variation in the S index according to soil clay content was able to be fitted using a double-linear-line approach, with decrease in the S index being much faster for clay content less than 20 %. In contrast, the bulk density index was found to be less important than clay content. The average S index was 0.077, indicating that soil physical quality in the Haihe River Basin was good.

Albedo and estimates of net radiation for green beans under polyethylene cover and field conditions

This paper describes the albedo (r) and estimates of net radiation and global solar irradiance for green beans crop (Phaseolus vulgaris L.), cultivated in greenhouse with cover of polyethylene and field conditions, in Botucatu, SP, Brazil (22º 54' S; 48º 27' W; 850 m). The solar global irradiance (Rg) and solar reflected radiation (Rr) were used to estimate the albedo through the ratio between Rr and Rg. The diurnal curves of albedo were obtained for days with clear sky and partially cloudy conditions, for different phenological stages of the crop. The albedo ranged with the solar elevation, the environment and the phenological stages. The cloudiness range have almost no influence on the albedo diurnal amount. The estimation of radiation were made by linear regression, using the global solar irradiance (Rg) and net short-waves radiation (Rc) as independent variables. All estimates of radiation showed better adjustment for specific phenological periods compared to the entire crop growing cycle. The net radiation in the greenhouse has been estimated by the global solar irradiance measured at field conditions.

Mapping of quantitative trait loci for thermosensitive genic male sterility in indica rice

The objective of this work was to select and use microsatellite markers, to map genomic regions associated with the genetic control of thermosensitive genic male sterility (TGMS) in rice. An F2 population, derived from the cross between fertile and TGMS indica lines, was used to construct a microsatellite-based genetic map of rice. The TGMS phenotype showed a continuous variation in the segregant population. A low level of segregation distortion was detected in the F2 (14.65%), whose cause was found to be zygotic selection. There was no evidence suggesting a cause-effect relationship between zygotic selection and the control of TGMS in this cross. A linkage map comprising 1,213.3 cM was constructed based on the segregation data of the F2 population. Ninety-five out of 116 microsatellite polymorphic markers were assembled into 11 linkage groups, with an average of 12.77 cM between two adjacent marker loci. The phenotypic and genotypic data allowed for the identification of three new quantitative trait loci (QTL) for thermosensitive genic male sterility in indica rice. Two of the QTL were mapped on chromosomes that, so far, have not been associated with the genetic control of the TGMS trait (chromosomes 1 and 12). The third QTL was mapped on chromosome 7, where a TGMS locus (tms2) has recently been mapped. Allelic tests will have to be developed, in order to clarify if the two regions are the same or not.