Macrofilaricidal and microfilaricidal effects of Neurolaena lobata, a Guatemalan medicinal plant, on Brugia pahangi.

Twelve extracts of 11 Guatemalan medicinal plants were initially screened in vitro for potential macrofilaricidal activity against Brugia pahangi, a lymphatic dwelling filarial worm, using concentrations from 125 to 1000 microg ml(-1) of each extract that could be dissolved in the culture medium. Of 12 extracts used, the ethanol extract of leaves of Neurolaena lobata showed the strongest activity against the motility of adult worms. Subsequently, the extract of N. lobata was extensively examined in vitro for macro- and micro-filaricidal effects using a series of concentrations of 500, 250, 100, 50 and 10 microg ml(-1). The effects were assessed by worm motility, microfilarial release by female worms and a MTT assay. The effect on the motility of adult worms was observed in a concentration- and time-dependent manner. The time required to stop motility of both sexes of adult worms was 6 h at 500 microg ml(-1), 24 h at 250 microg ml(-1), and 3 days for females and 4 days for males at 100 microg ml(-1). The movement of females ceased at 4 days at a concentration of 50 microg ml(-1) whereas the motility of males was only reduced. The loss of worm's viability was confirmed by the MTT assay and was similar to the motility results. These concentrations, including 10 microg ml(-1), prevented microfilarial release by females in a concentration- and time-dependent manner. Concentrations higher than 100 microg ml(-1) even induced mortality of the microfilariae. The present study suggested that the ethanol extract of Neurolaena lobata has potential macro- and micro-filaricidal activities.

The distribution of microfilariae in the skin of Guatemalan onchocerciasis patients: an evaluation of diagnostic potentials.

Quantitative examinations for the distribution and density of microfilariae were carried out on male Guatemalan onchocerciasis patients by taking 6 or 13 skin biopsies each from 74 subjects. We found that microfilariae were distributed most frequently on the iliac crest and secondarily in the scapular region; higher detection rates and greater microfilarial densities were seen in these areas. From these results we recommend the taking of biopsies from both of these anatomical regions so as to avoid false negative cases. The infected patients were classified into 3 groups (light, moderate and heavy infections) on the basis of the numbers of negative skin snips out of the 13 biopsies taken. In heavy infections, a higher densities of microfilariae were detected in the head and neck regions, as compared with the densities in the lower extremities (calf). The high concentration of microfilariae in these regions is of importance in relation to the development of eye lesions and/or transmission. Of patients who had microfilariae in the anterior segment of the eye some had negative skin biopsies frm the outer canthus, retroauricular region and neck.

Dermatological survey of onchocerciasis in Guatemala.

A comprehensive survey of onchocerciasis was performed on 1259 inhabitants in San Vicente Pacaya and the surrounding area. The total number of inhabitants examined was 1259, 763 males and 496 females. An examination of microfilarial density was carried out by the skin snip method. There were 610 microfilarial positives (50.4%) out of 1211 persons, 448 males (60.9%), and 162 females (34.1%). Nodules were examined by palpation, and there were 404 (32.8%) positives for nodule out of 1232 persons, 299 males (40.3%), and 105 females (21.4%). Eczematous dermatitis was seen in 149 persons, the generalized type in 13, and other types in 136. Depigmentation on the lower extremities was seen in 290 persons, the severity was graded from 0 to 4, and grade 1 depigmentation was seen in 91; grade 2, in 143; grade 3, in 52 and grade 4 in 4 persons out of 1159 individuals examined. The positives for lymphadenopathy (above grade 2) were 464 (41.2%) out of 1127 individuals. Itching was seen in 129 (18.3%) of 705 individuals examined by the inquiry card method. The correlation between skin changes and onchocerciasis was also analyzed. Those individuals who were microfilarial positive in the skin snip and eyes, and who had palpable nodules were defined as being onchocerciasis patients. There was a high frequency of eczematous dermatitis, depigmentation on the lower extremities and lymphadenopathy in the onchocerciasis infected group, compared with the non-infected group, but there was no difference in the rate of itching between them. Hanging groin, elephantiasis and "erisipela de la costa" were not seen in this endemic area, and the degree of skin changes was slight, compared with that reported in Africa thus far.

A sero-epidemiological study of onchocerciasis with the indirect hemagglutination test.

A sero-epidemiological study of onchocerciasis was carried out with the IHA test in Guatemala. In the endemic area, 94.8% of 191 subjects with microfilariae, 82.1% of 172 cases with onchocercal nodules, and 22.5% of 236 cases without either microfilariae or nodules gave positive reactions in the IHA test. On the other hand, the positive rate was 3.4% and 2.0% in each of 2 nonendemic populations. A follow-up study of reactors in the IHA with neither microfilariae nor nodules showed that 11 (42%) of 26 cases were microfilaria positive 6--7 mo after the first examination. A clear correlation was found between the results of the IHA test and skin biopsy in surveyed communities. The age distribution curve of the IHA test was closely associated with skin biopsy although the former was higher. IHA titers rose in proportion to microfilarial density. A sex-related difference was evident in the IHA positive rate in the subjects from medium and low endemic areas, but little difference was shown in individuals from a higher endemic area.

Onchocerciasis in San Vicente Pacaya, Guatemala.

An epidemiological survey for onchocerciasis was carried out in the San Vicente Pacaya area of Guatemala. A total of 2,153 inhabitants were examined by a single skin snip, and 664 (30.8%) were positive for microfilariae. Slit lamp examination of the anterior chamber of the eye revealed microfilariae in 6.2%; the positive rate for microfilariae in the anterior chamber was proportional to the microfilarial density in the skin. The altitude-dependent character of onchocercal infection was shown in this mountainous area, suggesting that transmission occurs principally between 600 and 1,300 m. Of, 1,217 persons examined simultaneously by skin snip and by palpation for nodules, 587 were positive by one or both methods; 101 infections were detected by onchocercal nodules only and these were usually in children or persons living in areas of low endemicity. It was concluded that both the skin snip and a search for nodules are required for accurately determining the prevalence of onchocerciasis in Guatemala.

The indirect hemagglutination test for onchocerciasis performed with blood collected on filter paper.

In Guatemala, an indirect hemagglutination (IHA) test for onchocerciasis, performed with a crude saline extract of Onchocerca volvulus and blood samples taken on filter paper, showed a high level of sensitivity and specificity. IHA titers of blood samples from the ear lobe taken on filter paper and of sera obtained by venipuncture showed a high correlation.