Importância do método de gota espessa de sangue no diagnóstico de filárias simpátricas no Amazonas, Brasil / The importance of thick blood film method for sympatric filarial diagnosis in Amazon basin

Esse trabalho relata os métodos de diagnósticos para filárias humanas no sangue, referindo-se a importância do método de gota espessa em relação aos métodos de filtração em membrana de policarbonato e Knott na diferenciação das espécies de microfilárias, principalmente em áreas de ocorrência de mais de uma espécie, como em regiões do Amazonas. Lâminas com microfilárias de Mansonella ozzardi foram montadas e fotografadas pelos diferentes métodos de diagnósticos. O método da gota espessa de sangue é o mais confiável, pois permite visualizar com nitidez o espaço cefálico e caudal, disposição dos núcleos caudais e formato da cauda, que são características morfológicas que diferenciam as espécies de microfilárias sanguíneas que ocorrem no Amazonas.

In this work, the visual diagnostic methods for human filarias in the blood are evaluated. The thick blood film method is more faithful to identify the microfilariae species if compared to policarbonate membrane filtration and Knott methods, especially in areas where more than one species occur, as in Amazon basin. Slides with Mansonella ozzardi microfilariae were mounted and photographed by different diagnosis methods. The thick blood film method is much easier to see the diagnostic morphological characters as cephalic and caudal space, pattern of nucleation of the tail and tail shape that can separate the blood Amazonian microfilariae species.

Evaluation of endocytic capacity and NADPH-oxidase activity from armadillo (Dasypus novemcinctus) eosinophils infected with microfilariae.

Endocytic activity of phagocytic cells from armadillos infected with viruses, parasites or bacteria is unknown. This report shows that eosinophils from armadillos infected with microfilaria act against these helmintic parasites but have deficiencies in their oxygen-dependent bacteriocidal mechanisms and also in endocytic capacity against yeast.

Blood parasites of two Costa Rican amphibians with comments on detection and microfilaria density associated with adult filarial worm intensity.

The 2 objectives of this study were: (1) to compare parasite detectability in blood smears obtained from toe-clips versus the heart from amphibian hosts; and (2) to test whether microfilariae density is correlated with adult filarial worm intensity. We examined blood parasites of 2 species of amphibians, Rana vaillanti (n = 45) and Eleutherodactylus fitzingeri (n = 36), from Costa Rica collected during the summer of 2003. Separate blood smears were obtained from toe-clips and the heart during necrospy. Eight species of blood parasites were identified from R. vaillanti and 1 from E. fitzingeri. Each parasite species was counted in a 2 x 2.2-cm2 area on each blood smear, and the density of host red blood cells (RBCs) was estimated using a sub-sampling approach, allowing parasite infections to be expressed as individuals per RBC. The detection failure rate for toe-cut smears ranged from 71-100% (x = 92.3%) and from 0-9% (x = 2.4%) for heart smears, depending on parasite species. The density of RBCs was significantly higher in smears produced from heart samples and may explain the differences in detectability. Foleyellides striatus microfilariae densities (per RBC) were significantly correlated with adult female worm intensity (R2 = 0.32, P = 0.011).

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.

Diagnosis of Wuchereria bancrofti infection by the polymerase chain reaction using urine and day blood samples from amicrofilaraemic patients.

A sensitive and specific polymerase chain reaction (PCR) based on a highly repeated deoxyribonucleic acid (DNA) sequence (188 bp; SspI repeat) was tested for the detection of Wuchereria bancrofti DNA in blood and urine samples collected during the day from individuals in Coque, Recife, Brazil, an endemic area for W. bancrofti. All microfilaraemic individuals were also positive by PCR, irrespective of the samples used. The PCR system was capable of detecting W. bancrofti DNA in amicrofilaraemic individuals: c. 93% were positive by PCR when day blood samples were used and 59.7% when urine samples collected at 07:00 were used. Thus, nocturnally periodic W. bancrofti infection can be detected in blood samples collected during the day, which is convenient for large-scale screening. In addition, non-invasive urine collection provided suitable samples for PCR, which is clearly advantageous for preliminary mass diagnosis.

Bancroftian filariasis in Maceio, state of Alagoas, Brazil: Observations on Culex quinquefasciatus after blood feeding on individuals with different densities of microfilariae in the peripheral blood stream.

Females of laboratory-reared Culex quinquefasciatus were allowed to take blood meals on individuals with low (1-100 microfilariae/ml of peripheral blood), moderate (101-500 microfilariae/ml), and high (> 500 microfilariae/ml) microfilaremia. The mosquitoes ingested 1.39-3.80 microl of blood and infective third-stage (L3) larvae were first recorded 13-14 days after the infecting blood meal. The number of microfilariae ingested by mosquitoes was proportional to the density of microfilariae in the peripheral blood of the human subjects, but with a concentration factor of up to 6.5 times the expected number. Survival of mosquitoes was not influenced by the density of microfilariae in the peripheral blood of infected individuals. Infectivity indices were proportional to microfilaremia in human subjects. The number of L3 larvae/female (intensity of infection) was not influenced by individual microfilaremia. The highest vector efficiency and the best experimental infection index were recorded in mosquitoes that blood fed on individuals with moderate microfilaremia. The results are discussed in relation to the transmission on Bancroftian filariasis in the study area.

Randomised placebo-controlled comparison of ivermectin and albendazole alone and in combination for Wuchereria bancrofti microfilaraemia in Haitian children.

BACKGROUND: Lymphatic filariasis and intestinal helminth infections are important disorders in tropical areas. Periodic treatment with albendazole is now used in many school-based intestinal helminth-control programmes. However, few such programmes exist for lymphatic filariasis, despite evidence that single-dose treatment with ivermectin can greatly reduce the concentration of Wuchereria bancrofti microfilariae in the blood for months to years. We aimed to assess the potential for school-based control of lymphatic filariasis by investigating the efficacy and tolerability or combined ivermectin and albendazole in Haitian schoolchildren. METHODS: In January, 1996, we collected 832 20 microL capillary blood samples for inclusion in a randomised controlled study from children aged 5-11 years, and examined them by microscopy for W bancrofti microfilariae. Infected children were randomly assigned treatment with placebo (n = 29), a single 200-400 micrograms/kg dose of ivermectin (mean, 273 micrograms/kg, n = 28), 400 mg albendazole (n = 29), or a combination of 200-400 micrograms/kg ivermectin and 400 mg albendazole (n = 24). Children with high concentrations of microfilariae in the blood were admitted to hospital and adverse reactions were monitored for 3-5 days, otherwise children were examined at school or during a visit to their home. 4 months after treatment, we examined blood samples again for microfilariae. FINDINGS: 113 microfilaraemic children were enrolled (mean age 7.8 years). 4 months after treatment, the proportion of children who remained positive for microfilariae was significantly lower in the ivermectin plus albendazole group (four [17%]), but there were no significant changes in the other three groups (20 [69%] placebo, 22 [76%] albendazole alone, 17 [61%] ivermectin alone remained positive; p = 0.004). Geometric mean microfilarial concentration decreased from 9.3 to 5.3 per 20 microL blood among children who received placebo; from 15.5 to 1.5 per 20 microL blood among those who received ivermectin only (p = 0.032); from 14.1 to 5.1 per 20 microL blood among those who received albendazole alone; and from 13.7 to 0.3 per 20 microL blood among those who received both ivermectin and albendazole (p = 0.0001). Systemic adverse reactions did not differ significantly between children who received ivermectin alone and those who were treated with ivermectin and albendazole [corrected]. INTERPRETATION: For children with W bancrofti microfilaraemia, combined treatment with ivermectin and albendazole was more effective than treatment with ivermectin only, with no measurable increase in severity of adverse reactions.

Evaluation of the Og4C3 ELISA in Wuchereria bancrofti infection: infected persons with undetectable or ultra-low microfilarial densities.

The recently developed Og4C3 ELISA, which detects circulating Wuchereria bancrofti antigen, appears promising for use in epidemiological surveys, but its sensitivity is unknown in persons with ultra-low microfilarial densities. We used the Og4C3 to test the sera of 282 persons who were microfilaria-positive in 1-16 ml of blood, 18 persons who were microfilaria-negative but who had ultrasonographic or biopsy evidence of adult W. bancrofti infection, and 63 lifelong residents of a non-endemic area of Brazil. A total of 276 (97.9%) persons with detectable microfilaraemia tested positive (optical density > 0.033). At microfilarial densities of < 1, 1-30, and > 30 microfilariae per ml of blood, the sensitivity of the Og4C3 was 72.2, 97.6 and 100%, respectively (chi 2-test for trend, P < 10(-6)). The assay was positive in 66.7% of amicrofilaraemic persons with evidence of adult worm infection and in one (1.6%) of 63 residents of the non-endemic area (specificity, 98.4%). Our findings support the increasingly widespread use of the Og4C3 for field investigations and epidemiological assessments. However, the sensitivity of the assay may be low in persons who are microfilaria-negative or with densities of < 1 microfilaria per ml.

Studies on the periodicity and intravascular distribution of Wuchereria bancrofti microfilariae in paired samples of capillary and venous blood from Recife, Brazil.

We examined the periodicity and intravascular distribution of Wuchereria bancrofti microfilariae (mf) and determined the effect of these parasite properties on the accuracy of blood filming and filtration methods for diagnosis of bancroftian filariasis in the endemic area of Recife, Brazil. Microfilariae in both venous and capillary blood exhibited a nocturnal periodicity pattern with a relatively high amplitude. Overall, capillary blood contained approximately 1.25 times the number of mf present at the same time in the same volume of venous blood. However, the ratio of mf present in capillary and venous blood varied over a 24-hour period, so that the fewest mf were present in the capillary bed of the skin at the time when biting activity of the local Culex vector is the lowest. Twenty or 60 microliters blood films did not reliably detect carriers with fewer than 100 or 60 mf/ml venous blood, respectively, and were thus inadequate for the identification of low density mf carriers. In contrast, all carriers with > 1 mf/20 or 60 microliters blood smear at night could be identified during daytime hours by filtration of 1 micromilligram venous blood.

Blood parasites in birds from Monteverde, Costa Rica.

In a survey of avian blood parasites in Costa Rica, 51 (11%) of 479 birds sampled were infected by at least one species of hematozoan. Fourteen of the 60 species of birds in the survey were examined for the first time. Infections were most common in ramphastids and emberizids and infrequent in other taxa. Among resident species, infections were more commonly detected during the wet season when most birds breed than during the dry season when few birds breed. Infections caused by Haemoproteus sp. were most common, while Plasmodium sp., Leucocytozoon sp., Trypanosoma sp., and microfilarial infections were rare. The intensity of the 40 Haemoproteus infections in adult birds was low, with a mean +/- SE of 12.5 +/- 3.7 infected cells per 10,000. Haemoproteus infections did not undergo seasonal changes in intensity.

Antifilarial cellular responses detected in a Haitian pediatric population by use of a microblastogenesis assay.

Previous reports have demonstrated age-related shifts in antifilarial humoral immune responses in 6- to 10-year-old Haitian children; the responses consisted of elevated parasite-specific IgG2 and IgG3 in amicrofilaremic children and elevated IgG4 in microfilaremic children. In this study, the cell-mediated immune responses to soluble adult and microfilarial extracts of Brugia pahangi, determined by use of a microblastogenesis assay, were examined. Capillary blood samples were collected by finger prick from 176 Haitian children in an area with endemic Wuchereria bancrofti. Antigen-specific cellular responsiveness varied as a function of infection status but not age or sex; amicrofilaremic children had significantly greater responses to adult antigens than did microfilaremic children. Significant responses were detected in children less than 2 years of age; thus, correlations observed between filarial antigen-specific responses and infection status are established early in life.

[Lymphatic filariasis in Haiti. Results of treatment with diethylcarbamazine in the town of Limbe]. / Filariose lymphatique en Haiti. Résultats du traitement par le diéthylcarbamazine dans la ville du Limbe.

In the town of Limbe, in Haiti, 16,8% of the inhabitants show W. bancrofti microfilariae in their blood taken by finger prick between 8.00 and 10.00 p.m. From those positive patients, 208 have received a cure of DEC at the dosage of 6 mg/kg/day during ten days. The control of the therapeutic efficacity among 190 patients, after three and a half months of this cure, shows a negativity of 72%. The positives patients show a very low parasitic charge (MfD50 = 1.9). If the secondary effects are frequent (83.7%), they are often benign and temporary.

Filariasis en Colombia: presencia de Dipetalonema perstans en la Comisaria del Guainia / Filariasis in Colombia: presence of Dipetalonema perstans in the Comisaria del Guainia

Al examinar 75 muestras de sangre (metodo de Knott) colectadas en Puerto Inirida, Coco y Pajuil en la Comisaria del Guainia, Colombia, se descubrieron 26 portadores de micofilarias. En 18 personas habia unicamente Mansonella ozzardi; 3 tenian infeccion con M. ozzardi; 3 tenian infeccion con M. ozzardi y Dipetalonema perstans; y cinco solo con D. perstans. La infeccion por M. ozzardi se observo en blancos y en indigenas de las tribus curripacos, puinaves, tukanos, guananos y salivas, pero D. perstans se encontro solamente entre los curripacos. El numero de microfilarias (mf) circulantes fue bajo, 73% de los portadores tenian menos de 200 mf/ml de sangre; las personas que presentaron unicamente D. perstans tenian menos de 310 mf/ml. Estos resultados confirman la presencia de D. perstans en Colombia y sugieren que su prevalencia y distribucion en la Comisaria del Guainia y areas cercanas pueden ser mas altas de lo que se ha sospechado hasta el momento

Microfilaria bolivarensis: a new species of filaria from man in Venezuela.

An unsheathed microfilaria, unlike any other already described from man, was found in thick blood smears and Knott samples taken from Amerinds living along the upper Caura River in Bolivar State, Venezuela. The mirofilaria, which measures 256 x 7-8 micrometers in thick blood smears and 300 x 8 micrometers in Knott samples, is easily differentiated morphologically from Mansonella ozzardi with which it often occurs concomitantly. It is readily differentiated, on morphological criteria, from all other species of filariae which infect man, as well. The parasite has been named Microfilaria bolivarensis after the geographical locality in which it was discovered.

[The use of the Knott technic in an endemic area of filariasis in Puerto Limon, Costa Rica]. / El uso de la técnica de Knott en una zona endémica de filariasis en Puerto Limón, Costa Rica.

One hundred and nineteen people living in the Roosevelt county of Puerto Limón were submitted to the Knott technique for diagnosing microfilaremia bancrofti. This was the first time the technique was applied in Costa Rica. A 17.6% positivity was obtained. Some considerations on filariasis spreading to other regions of the country with the possibility of outbreaks of tropical eosinophilia or filariasis without microfilaremia are made.

The association of blood groups, ABH secretion, haptoglobins and hemoglobins with filariasis.

Results on 605 patients and 597 controls carefully matched in relation to sex, age, marital status and profession are reported. A higher frequency of nonsecretors, as well as an excess of B and lack of A2 were observed among the filarial patients. Since these differences do not occur in other series, however, their significance is doubtful.