Halipegus dubius Klein, 1905 (Trematoda, Hemiuridae): a redescription, with notes on the working of the ovarian complex

A redescription of Halipegus dubius Klein, 1905, of Brazilian frogs Leptodactylus pentadactylus and L. ocellatus, is presented. The parasite was found exclusively in the buccal cavity, in number varying from 1 (in most instances) to 3, and exceptionally 25 (one instance). Morphological data were based on 40 whole-mounted specimens and 4 serially sectioned ones. Larval development takes place in planorbid snails (Biomphalaria glabrata and B. tenagophila) and in an undetermined species of cyclops. Working of the ovarian complex was followed by microscopical observation of life specimens. Constrating with the similarity of the four American species of Halipegus currently recognized as valid, they can be easily separated by the characters of their cercariae.

Evaluation and additional recommendations for preparing a whole blood control material

OBJECTIVE: The assessment of an easy to prepare and low cost control material for Hematology, available for manual and automated methods. MATERIAL AND METHOD: Aliquots of stabilized whole blood were prepared by partial fixation with aldehydes; the stability at different temperatures (4. 20 and 37 °C) during periods of up to 8-9 weeks and aliquot variability with both methods were controlled. RESULTS: Aliquot variability with automated methods at day 1, expressed as CV% (coefficient of variation) was: white blood cells (WBC) 2.7, red blood cells (RBC) 0.7, hemoglobin (Hb) 0.6, hematocrit (Hct) 0.7, mean cell volume (MCV) 0.3, mean cell hemoglobin (MCH) 0.6, mean cell hemoglobin concentration (MCHC) 0.7, and platelets (PLT) 4.6. The CV (coefficient of variation) percentages obtained with manual methods in one of the batches were: WBC 23, Hct 2.8, Hb 4.5, MCHC 5.9, PLT 41. Samples stored at 4ºC and 20ºC showed good stability, only a very low initial hemolysis being observed, whereas those stored at 37ºC deteriobed a rapidly (metahemoglobin formation, aggregation of WBC and platelets, as well as alteration of erythrocyte indexes). CONCLUSIONS: It was confirmed that, as long as there is no exposure to high temperatures during distribution, this material is stable, allowing assessment, both esternal and internal, for control purposes, with acceptable reproductivity, both for manual and auttomatic methods.

Field evaluation of the whole blood immunochromatographic test for rapid bancroftian filariasis diagnosis in the northeast of Brazil

This study evaluated the whole blood immunochromatographic card test (ICT card test) in a survey performed in Northeastern Brazil. 625 people were examined by the thick blood film (TBF) and ICT card test. Residents of a non-endemic area were also tested by the whole blood card test and Og4C3. The sensitivity of the ICT card test was 94.7% overall, but lower in females than males, based on the reasonable assumption that TBF is 100% specific. However, since TBF and other methods have unknown sensitivity, the true specificity of the card test is unknown. Nevertheless, it is possible to estimate upper and lower limits for the specificity, and relate it to the prevalence of the disease. In the endemic area, the possible range of the specificity was from 72.4% to 100%. 29.6% of the card tests performed in the non-endemic area exhibited faint lines that were interpreted as positives. Characteristics of the method including high sensitivity, promptness and simplicity justify its use for screening of filariasis. However, detailed information about the correct interpretation in case of extremely faint lines is essential. Further studies designed to consider problems arising from imperfect standards are necessary, as is a sounder diagnostic definition for the card test.

Refractive index matching applied to fecal smear clearing

Thick smears of human feces can be made adequate for identification of helminth eggs by means of refractive index matching. Although this effect can be obtained by simply spreading a fleck of feces on a microscope slide, a glycerol solution has been routinely used to this end. Aiming at practicability, a new quantitative technique has been developed. To enhance both sharpness and contrast of the images, a sucrose solution (refractive index = 1.49) is used, which reduces the effect of light-scattering particulates. To each slide a template-measured (38.5 mm³) fecal sample is transferred. Thus, egg counts and sensitivity evaluations are easily made.

Evaluation of the human immunodeficiency virus type 1 and 2 antibodies detection in dried whole blood spots (dbs) samples

Human Immunodeficiency Vírus Type 1 and 2 antibodies detection was performed in 457 dried whole blood spots samples (S&S 903). Q-Preven HIV 1+2 was the screening test used. The results were compared with the gold standard serum tests by ELISA (Cobas Core e Axsym HIV1/2 gO) and imunofluorescence was the definitive confirmatory test. The samples were obtained from the Hospital Nossa Senhora da Conceição in Porto Alegre, RS - Brazil, through whole blood transfer to filter paper card and sent to Caxias do Sul, RS - Brazil where the tests were performed. The dried whole blood spot stability was evaluated with two different panels. The first one was composed of five negative and five positive samples stored at room temperature, 4 ºC, -20 ºC and -70 ºC, while the second was composed of two negative and three positive samples stored at 37 ºC (humidity <50%). Each sample was screened every week for six weeks. These measurement results didn't show variation during the study period. The detected sensibility was 100%, specificity was 99.6%, the positive predictive value was 99.5% and negative predictive values were 100%. The results demonstrated high performance characteristics, opening a new perspective of dried whole blood spot utilization in HIV screening diagnosis.

Production of cytokine and chemokines by human mononuclear cells and whole blood cells after infection with Trypanosoma cruzi

INTRODUCTION: The innate immune response is the first mechanism of protection against Trypanosoma cruzi, and the interaction of inflammatory cells with parasite molecules may activate this response and modulate the adaptive immune system. This study aimed to analyze the levels of cytokines and chemokines synthesized by the whole blood cells (WBC) and peripheral blood mononuclear cells (PBMC) of individuals seronegative for Chagas disease after interaction with live T. cruzi trypomastigotes. METHODS: IL-12, IL-10, TNF-α, TGF-β, CCL-5, CCL-2, CCL-3, and CXCL-9 were measured by ELISA. Nitrite was determined by the Griess method. RESULTS: IL-10 was produced at high levels by WBC compared with PBMC, even after incubation with live trypomastigotes. Production of TNF-α by both PBMC and WBC was significantly higher after stimulation with trypomastigotes. Only PBMC produced significantly higher levels of IL-12 after parasite stimulation. Stimulation of cultures with trypomastigotes induced an increase of CXCL-9 levels produced by WBC. Nitrite levels produced by PBMC increased after the addition of parasites to the culture. CONCLUSIONS: Surface molecules of T. cruzi may induce the production of cytokines and chemokines by cells of the innate immune system through the activation of specific receptors not evaluated in this experiment. The ability to induce IL-12 and TNF-α contributes to shift the adaptive response towards a Th1 profile.

Climatic aspects of the Amazonian Tropical Forest

The purpose of this study is to analyse the climatic aspects of the data collected in a forest site in comparison with conventional data obtained at different sites, such as clearing, rural an urban areas. The results showed that diverse climatic conditions do exist among the sites: the urban site showed higher temperature and lower relative humidity. In addition, evapotranspiration (potential and actual rates) was computed from the forest data set, using the classical Penman-Monteith's equation. The actual evapotranspiration is 30% of the potential value during dry period and seems to be almost constant during the whole year (tipically 2.0 to 2.5 mm day-1).

Clearing factor pancreático

Dosou-se o "clearing factor" (lipase lipoproteica), em pâncreas de ratos normais após serem submetidos a algumas condições experimentais. As seguintes conclusões podem ser extraídas: a) O pãncreas é o órgão mais rico nesta enzima (8 vezes mais que o tecido adiposo, até então considerado o mais rico em CF); b) O jejum total de 48 horas diminui significamente a atividade de CF; c) O complamin, o derivado do ácido nicotínico, agindo sobre a arteriosclerose experimental, não afeta a atividade da enzima; d) O ácido épsilon-aminocapróico (AEAC) potente antagonista da heparina, inibe a ativdade do Cf pancreático; e) Ratos com arteriosclerose experimental apresentam níveis diminuídos desta enzima no pâncreas.

Activity of Tabanids (Insecta: Diptera: Tabanidae) Attacking the Reptiles Caiman crocodilus (Linn.) (Alligatoridae) and Eunectes murinus (Linn.) (Boidae), in the Central Amazon, Brazil

Tabanid females are better known as hematophagous on man and other mammals, and linked to mechanical transmission of parasites. The association between tabanids and reptiles is poorly known, but has been gaining more corroboration through experiments and occasional observation in the tropics. The present study was conducted at a military base (CIGS/BI-2), situated 54 km from Manaus, Amazonas, in a small stream in a clearing (02°45'33"S; 59°51'03"W). Observations were made monthly, from April 1997 to March 1998, during two consecutive days. At the same time, other vertebrate animals were offered, including humans. However in this paper only data obtained on a common caiman, Caiman crocodilus (Linn.), and an anaconda, Eunectes murinus (Linn.), in diurnal observations from 05:30 a.m. to 18:30 p.m., will be discussed. A total of 254 tabanid specimens were collected, 40 from the anaconda and 214 from the caiman. Four tabanid species were recorded on these two reptiles: Stenotabanus cretatus Fairchild, S. bequaerti Rafael et al., Phaeotabanus nigriflavus (Kröber) and Tabanus occidentalis Linn. Diurnal activities showed species-specific patterns. The first three species occurred only in the dry season. T. occidentalis occurred during the whole observation period, and with increased frequency at the end of the dry season. We observed preferences for body area and related behavior of the host. Observations on the attack of tabanids on one dead caiman are also presented.

Preparation of sand fly (Diptera: Psychodidae: Phlebotominae) specimens for histological studies

Phlebotominae sand fly specimens were prepared for histological and physiological studies. Different fixatives were tested on sectioned and whole bodied adult females in order to obtain good fixation and provide satisfactory penetration of the embedding media. All fixed specimens were infiltrated (up to seven days under 5ºC) and embedded in hydroxyethyl metacrylate. Two-three µm sections were stained, mounted in Canada balsam and observed by light microscopy. Best results were achieved when whole bodied insects were double fixed in Bouin's and Carnoy's fluids (4 h/2 h) and stained in Hematoxilin/Eosin or fixed in calcium formaldehyde and stained in mercury bromophenol blue.

Whole blood assay to access T cell-immune responses to Mycobacterium tuberculosis antigens in healthy Brazilian individuals

The production of interferon gamma (IFNgamma) guarantees effective T cell-mediated immunity against Mycobacterium tuberculosis infection. In the present study, we simply compare the in vitro immune responses to Mycobacterium antigens in terms of IFNg production in a total of 10 healthy Brazilian volunteers. Whole blood and mononuclear cells were cultivated in parallel with PPD, Ag85B, and M. bovis hsp65, and five-days supernatants were harvested for cytokine detection by ELISA. The inter-assay result was that the overall profile of agreement in response to antigens was highly correlated (r² = 0.9266; p = 0.0102). Potential analysis is in current progress to dictate the usefulness of this method to access the immune responses also in tuberculosis patients and its contacts.

Four whole-istic aspects of schistosome granuloma biology: fractal arrangement, internal regulation, autopoietic component and closure

This paper centers on some whole-istic organizational and functional aspects of hepatic Schistosoma mansoni granuloma, which is an extremely complex system. First, it structurally develops a collagenic topology, originated bidirectionally from an inward and outward assembly of growth units. Inward growth appears to be originated from myofibroblasts derived from small portal vessel around intravascular entrapped eggs, while outward growth arises from hepatic stellate cells. The auto-assembly of the growth units defines the three-dimensional scaffold of the schistosome granulomas. The granuloma surface irregularity and its border presented fractal dimension equal to 1.58. Second, it is internally regulated by intricate networks of immuneneuroendocrine stimuli orchestrated by leptin and leptin receptors, substance P and Vasoactive intestinal peptide. Third, it can reach the population of ± 40,000 cells and presents an autopoietic component evidenced by internal proliferation (Ki-67+ Cells), and by expression of c-Kit+ Cells, leptin and leptin receptor (Ob-R), granulocyte-colony stimulating factor (G-CSF-R), and erythropoietin (Epo-R) receptors. Fourth, the granulomas cells are intimately connected by pan-cadherins, occludin and connexin-43, building a state of closing (granuloma closure). In conclusion, the granuloma is characterized by transitory stages in such a way that its organized structure emerges as a global property which is greater than the sum of actions of its individual cells and extracellular matrix components.

Evaluation of buffy coat 16S rRNA PCR, buffy coat culture and whole blood PCR for detection of bacteraemia

The use of Gram type-specific PCR on buffy coat from clinical specimens for the detection of bacteraemia was evaluated for the first time using whole blood culture as the gold standard. In addition, the established buffy coat culture and whole blood PCR were also compared. Gram-positive bacteria belonging to six species and Gram-negative bacteria from 10 species were isolated and identified by culture and detected using broad-range 16S rDNA primers and Gram-specific primers. Data from the three methods all conferred very high sensitivity, specificity, positive and negative predictive values when compared to whole blood culture. The Kappa coefficients of agreement were 0.9819 (buffy coat PCR), 0.9458 (whole blood PCR) and 1.0 (buffy coat culture), which establishes their validity as alternative methods to routine blood culture in detecting bacteraemia. In addition, results showed that there was a direct correlation of WBC counts greater than 12,000 cells per mm³ to the occurrence of bacteraemia as detected by the four methods (p < 0.05).

An indirect immunofluorescence antibody test employing whole eggs as the antigen for the diagnosis of abdominal angiostrongyliasis

Abdominal angiostrongyliasis is a potentially fatal zoonotic disease with a broad geographical distribution throughout Central and South America. This study assessed the performance of Angiostrongylus costaricensis eggs as the antigen in an indirect immunofluorescence assay for the determination of parasite-specific IgG and IgG1 antibodies. For prevalence studies, an IgG antibody titre > 16 was identified as the diagnostic threshold with the best performance, providing 93.7% sensitivity and 84.6% specificity. Cross reactivity was evaluated with 65 additional samples from patients with other known parasitic infections. Cross reactivity was observed only in samples from individuals infected with Strongyloides stercoralis. For clinical diagnosis, we recommend the determination of IgG only as a screening test. IgG1 determination may be used to increase the specificity of the results for patients with a positive screening test.

Whole-cell antigenicity data support sequence-based kinetoplastid taxonomy

Early immunological data, obtained by immunodiffusion and immunoelectrophoresis, on the whole-cell antigenicity of kinetoplastid protozoa were retrieved and used to construct a dendrogram of antigenic distances. Remarkably, they supported the same taxonomic conclusions as analyses based on DNA and protein sequence data.