Abraham Rocha

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 (χ2‐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.

Ultrasonographic assessment of the adulticidal efficacy of repeat high-dose ivermectin in bancroftian filariasis.

Since diethylcarbamazine, the drug recommended for treatment of lymphatic filariasis, seems only partially effective against the adult worm, intense interest persists in identifying a macrofilaricidal drug for this infection. To evaluate directly in vivo the macrofilaricidal activity of repeat high‐dose ivermectin, 15 men who had living adult Wucbereria bancrofti detected in the scrotal area by ultrasound were treated with 400 μg/kg of ivermectin at 2‐week intervals for 6 months (total dose, 4.8 mg/kg). Serial ultrasound examinations were performed before, during, and for 6 months after treatment. Profound suppression of microfilaraemia followed the first dose of ivermectin, but movements characteristic of the adult worm on ultrasound remained unchanged both in location and pattern. Even when given in total doses of 4.8 mg/kg, ivermectin appears to have no observable activity against adult W. bancrofti, although its ability to suppress microfilaraemia makes it potentially useful for the control of lymphatic filariasis.

Treatment of bancroftian filariasis in Recife, Brazil: a two-year comparative study of the efficacy of single treatments with ivermectin or diethylcarbamazine

The effectiveness of single oral doses of ivermectin (200 or 400 micrograms/kg) and diethylcarbamazine (DEC, 6 mg/kg), preceded 4 d earlier by either placebo or very small doses of these drugs, was compared, over a 2-year period, in a double-blind trial in 67 microfilaraemic Brazilian men with bancroftian filariasis. Regimens containing ivermectin alone decreased the number of microfilariae significantly faster and more effectively for the first month after treatment than regimens containing DEC alone, but the latter were significantly more effective throughout the second year after treatment (1.7-8.2% of pretreatment levels with DEC vs. 12.6-30.8% with ivermectin during that period); the higher ivermectin dose showed a tendency towards more effectiveness than the lower dose. Most effective was the combination of ivermectin (20 micrograms/kg) followed 4 d later by DEC (6 mg/kg), with reduction of microfilaraemia to 2.4% of pretreatment levels at 2 years. Adverse reactions were well tolerated with all regimens, the reactions being significantly more generalized (i.e., fever) following ivermectin and localized (i.e., scrotal inflammatory nodules around dying adult worms) following DEC. Further trials of single-dose combination therapy vs. single high doses of ivermectin or DEC should determine the ideal regimen for treatment and control of bancroftian filariasis.

Comparative efficacy of three different diethylcarbamazine regimens in lymphatic filariasis.

To assess the efficacy of diethylcarbamazine (DEC) in clearing Wuchereria bancrofti microfilariae (mf) from the circulation, we conducted a single blind hospital-based therapeutic trial of 3 DEC regimens. All patients were assessed by filtration of 1 mL of venous blood taken before and 1, 3, 6 and 12 months after DEC administration. The efficacy of a 12 d course of 6 mg/kg DEC once daily was identical to that of a similar course with 2 mg/kg given 3 times daily, indicating that split-dose treatment does not improve mf clearance over single daily drug administration. Microfilarial densities in patients treated only once with 6 mg/kg DEC remained significantly higher at 1, 3, and 6 months after treatment. However, all 3 treatment regimens proved equally effective in controlling microfilaraemia after 12 months, when 41, 42 and 40% of patients in the 3 treatment groups were amicrofilaraemic. These results suggest that a single DEC dose of 6 mg/kg administered annually or biannually may be a suitable regimen to control bancroftian filariasis in Recife, Brazil.

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 ˜ 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 μl 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 > I mf/20 or 60 μl blood smear at night could be identified during daytime hours by filtration of I ml venous blood

Histological evidence for adulticidal effect of low doses of diethylcarbamazine in bancroftian filariasis

The ability of diethylcarbamazine (DEC) to kill adult Wuchereria bancrofti worms was evaluated by examining lymphatic nodules formed after treatment with 4 different treatment schedules of 193 males living in the endemic area of Greater Recife, Brazil. Lymphatic nodules appeared in the spermatic cord or upper extremities in 43 of 138 microfilaraemic individuals, in 3 of 30 amicrofilaraemic patients with filarial disease manifestations, and in 1 of 25 asymptomatic amicrofilaraemic residents of the endemic area treated with DEC. Fourteen of these nodules were surgically removed 10-150 d after the start of treatment. Regardless of the DEC dosage and schedule used, all nodules contained damaged and degenerating adult worms. An exuberant granulomatous process with large numbers of eosinophils and progressive fibrosis gradually developed around the dead parasites. The mechanism(s) by which DEC killed adult W. bancrofti could not be determined.

Epidemiological study of bancroftian filariasis in Recife, Northeastern Brazil

Wuchereria bancrofti in Pernambuco was first documented in 1952 (Azevedo & Dobbin 1952), and since then it has been reported in surveys carried out in selected areas of Recife. Several surveys were carried out from 1981 to 1991 by SUCAM. In the 1985 SUCAMs report the disease is considered under control. The CPqAM Filariasis Research Program was established in 1985 and a filarial survey was carried out in the town of Olinda, Greater Recife. In order to verify the real epidemiological situation, a study was conducted in the city of Recife. 21/36 of the Special Zones of Social Interest (ZEIS), were randomly selected for the present study. From 10,664 persons screened, 683 were positive and the prevalence rate for microfilaraemia (mf) varied from 0.6% to 14.9%. A mean mf prevalence of 6.5%, showed that the infection occurs in a wide geographic distribution in Greater Recife and that the intensity of transmission is a real and potential threat to public health in affected communities. Mf rate among males and females differed significantly. Due to the rapid increase in population, unplanned urban settlements, poor sanitary facilities and the favorable geographical conditions to the development of the vector, filariasis may actually be increasing in Recife.

Comparison of tests for the detection of circulating filarial antigen (Og4C3-ELISA and AD12-ICT) and ultrasound in diagnosis of lymphatic filariasis in individuals with microfilariae

Significant advances were made in the diagnosis of filariasis in the 1990s with the emergence of three new alternative tools: ultrasound and tests to detect circulating antigen using two monoclonal antibodies, Og4C3 and AD12-ICT-card. This study aimed to identify which of these methods is the most sensitive for diagnosis of infection. A total of 256 individuals, all male and carrying microfilariae (1-15,679 MF/mL), diagnosed by nocturnal venous blood samples, were tested by all three techniques. The tests for circulating filarial antigen concurred 100% and correctly identified 246/256 (96.69%) of the positive individuals, while ultrasound detected only 186/256 (73.44%). Of the circulating antigen tests, ICT-card was the most convenient method for identification of Wuchereria bancrofti carriers. It was easy to perform, practical and quick.

Comparative studies on endemic areas of bancroftian filariasis in Greater Recife, Brazil

Two districts in Recife (Santo Amaro and Campo Grande) and two districts in Olinda (Sapucaia and Salgadinho), were selected for a comparative study of bancroftian filariasis in Greater Recife. Selection parameters included similar socio-economic, demographic, and endemic levels of lymphatic filariasis. In the districts studied, streets were chosen randomly. These clusters consisted of 110 people each. A population sample was stratified by sex and age: 0-4, 5-9, 10-14, 15-19, 20-29, 30-39, 40-49, 50-59 and 60 years old and over. The parasitological data were obtained by measuring thick blood smears (60 micro l), collected from 8:00 to 12:00 PM and processed and stained with hematoxylin. The data were described in tables, and logarithimic expression graphics were used to analyze parasitic densities. In general, Recife showed a prevalence of 13.5%, and Olinda 12.3%. Mean parasitic densities were 41 and 70 microfilariae/60 micro l (mf/60 micro l), respectively. Higher microfilaremic rates were observed in Recife for the 20-29-year age bracket and in Olinda in the 30-39-year bracket. The authors concluded that the disease has returned to former endemic levels, and the data described call for a reevaluation of control campaigns carried out by the Brazilian National Health Foundation, since the endemic has already reached serious proportions.

Circulating filarial antigen in the hydrocele fluid from individuals living in a bancroftian filariasis area - Recife, Brazil: detected by the monoclonal antibody Og4C3-assay

The purpose of this study was to examine the circulating filarial antigen (CFA) detected by the monoclonal antibody (mAb) Og4C3-ELISA in paired samples of serum and hydrocele fluid from 104 men with hydrocele, living in an endemic area of Wuchereria bancrofti. Nocturnal blood specimens were filtered and examined for microfilariae (MF) and ultrasound was used in order to identify the presence of adult worms (the filaria dance sign - FDS) in the lymphatic vessels of the scrotal area. Four groups were selected according to their parasitological status: group I - 71 MF- and FDS-; group II - 21 MF+ and FDS+; group III - 10 MF- and FDS+ and group IV- 2 MF+ and FDS-. CFA was identified simultaneously (fluid and serum) in 11 (15.5%), 21 (100%), 3 (30%), and 1 (50%) in groups I, II, III, and IV, respectively. In despite of high CFA+ level (antigen Og4C3) units/ml, the Geometrical Mean (GM) = 2696) in the sera of these 36/104 paired samples, when compared to the hydrocele fluid, (GM = 1079), showed a very good correlation between the CFA level in the serum and CFA level in the fluid (r = 0.731). CFA level in the serum of the 23 microfilaremics (groups II and IV) was extremely high (GM = 4189) and was correlated with MF density (r = 0.442). These findings report for the first time the potential alternative use of the hydrocele fluid to investigate CFA using the mAb Og4C3-ELISA.