Onchocerca-Simulium complexes in Venezuela: can human onchocerciasis spread outside its present endemic areas?

The compatibility between sympatric and allopatric combinations of Onchocerca volvulus-anthropophilic species of Simulium was studied in the north-eastern focus of human onchocerciasis as well as in a densely populated locality of the Amazonas State in Venezuela. The objectives were to test the conjecture that local adaptation exists between the parasite and its vectors (the Onchocerca-Simulium complex hypothesis), and assess the possibility of the infection spreading from its present distributional range. For the homologous combination, O. volvulus-S. metallicum cytospecies E in Anzoátegui State (north-eastern focus), parasite yield was 45% in contrast to 1% for the heterologous, southern parasite-S. metallicum infection. This was significantly lower than the parasite yield (4-10%) expected after allowing for the effect of density-dependent limitation of infective larval output described in this paper for S. metallicum. The population of S. exiguum s.l. from southern Venezuela allowed no larval development beyond the L1 stage of either northern or southern parasites. Mechanisms for such refractoriness probably operate at the level of the thoracic muscles, not affecting microfilarial uptake or migration out of the bloodmeal. The parasite yield of southern O. volvulus in S. oyapockense s.l. flies biting man at Puerto Ayacucho (Amazonas) was about 1%, in agreement with the figures recorded for highly compatible sympatric combinations such as O. volvulus-S. ochraceum s.l. in Guatemala. No infective larval development of the northern parasite was observed in southern S. oyapockense. These results, together with considerations of typical worm burdens in the human host, presence/absence of armed cibaria in the simuliids, parasite-induced vector mortality, and fly biting rates, suggest a lower potential for onchocerciasis to spread between the northern and southern endemic areas of Venezuela than that between Amazonian hyperendemic locations and settlements outside this focus with high densities of S. oyapockense s.l.

Risk factors associated with human cystic echinococcosis in Florida, Uruguay: results of a mass screening study using ultrasound and serology.

Sonographic evidence of asymptomatic Echinococcus granulosus lesions in the liver was found in 156 of 9,515 persons in the Department of Florida, Uruguay. The sensitivity of ELISA and latex agglutination serology compared with ultrasound was 47.6% and 28.1%, respectively, and specificity was > 85%. There was a significant positive association between positive sonography and a personal history of previous but treated Echinococcus infection while those that were seropositive but ultrasound-negative were significantly more likely to have a personal history of infection or a history of infection in their family. Prevalence of infection increased significantly with age. There was no correlation between echinococcosis and dog ownership or home slaughter of sheep but offal disposal was important, with an increased prevalence of infection of 3.2%, 2.8%, and 3.1%, respectively, in persons feeding offal to dogs or burying or burning it compared with a prevalence of 0.8-1.5% in those using other methods of disposal. Almost half the population, when questioned, seemed to have sound knowledge about E. granulosus and described correct treatment of E. granulosus in dogs but this did not affect prevalence. There was a significant positive association between infection and the presence of a fenced fruit/vegetable garden and use of rural waters, particularly the cachimba (a small dam) and the aljibe (a cistern or tank) that collect rainwater from the ground surface and roofs, respectively.

Principles of the surgical approach in human liver cystic echinococcosis.

Ultrasonography (USG) has recently contributed much to the diagnostic of hepatic cystic echinococcosis (CE). The use of portable ultrasonograph allowed us to perform a community survey among 9482 people living in a high risk area for CE in the Florida Department (Uruguay). Positive USG results were found in 123 asymptomatic patients. 48 out of 51 USG positive cases were surgically confirmed and 3 were found to be false positive. The results of this survey allowed us to propose a new classification of the echographic imaging based on the parasite's various evolutive and involutive stages. The Echinococcus granulosus cyst size was compared with the parasite's evolutive stages. The cyst's segmentary topography and the related risk of CE is evaluated. The importance of cystic-biliary communication is pointed out and its rational surgical treatment described. Finally, an algorithm is presented facilitating the choice of a rational treatment.

Specific detection of human antibodies to Onchocerca volvulus.

Specific diagnosis of antibodies to Onchocerca was achieved through (1) the construction of direct and indirect ELISA systems, and (2) restricting ELISA assays to the IgG4 class. The direct ELISA was based on the isolation of a surface derived, low molecular weight surface antigen preparation containing two main antigens (M. wt. 16.2 and 12.8 kDA) as defined by Western blot analysis. The direct ELISA system detected antibodies in children of six years old, and may therefore be applicable to detecting reinvasion in OCP areas of Onchocerca volvulus control. The indirect ELISA system was a competitive binding ELISA-based assay using a monoclonal antibody recognising two Onchocerca components (M. wts. 15.6 and 25.9) on a Western blot. The direct and indirect ELISA systems were similarly specific and sensitive when evaluated in a preliminary survey. The direct ELISA system yielded a specificity and sensitivity of: 100% and 100% respectively, using Mexican endemic and Mexican intestinal nematode infection sera as positive and negative controls respectively: 91% and 96% respectively, using Venezuelan endemic and Venezuelan Mansonella ozzardi infection sera as positive and negative controls, respectively: 87% and 93% respectively, using African endemic and Papuan (New Guinea) Wuchereria bancrofti infection sera as positive and negative controls respectively: 93% and 93% respectively, using African endemic and Indian W. bancrofti infection sera as positive and negative controls respectively. Similar specificity and sensitivity levels were obtained when the same comparisons were made using the indirect (inhibition) ELISA assay. These values may be contrasted with the currently used PBS extract of O. volvulus which yielded specificities of less than 10% in all the above comparisons.(ABSTRACT TRUNCATED AT 250 WORDS)

Susceptibility of Plasmodium falciparum strains to chloroquine and mefloquine in the Amazonas Federal Territory of Venezuela.

The susceptibilities of 27 Plasmodium falciparum strains to chloroquine and mefloquine were studied in the area of Puerto Ayacucho, Amazonas Federal Territory of Venezuela, to determine their levels of resistance in vivo and in vitro. 50% of these strains showed chloroquine resistance in vivo. No grade III chloroquine resistance was found. 25% of the strains were resistant to chloroquine in vitro and 9% were resistant to mefloquine in vitro. Preliminary results suggest that strains resistant to Fansidar may also be found.

Sporotrichosis in the Orinoco river basin of Venezuela and Colombia.

Six cases of sporotrichosis from the Orinoco river basin of Venezuela and Colombia are described; two are of the localized cutaneous type and four are lymphocutaneous. Diagnosis was based on the patient's clinical history and mycological culture. Epidemiology and distinctive cultural habits of the patients are discussed in connection with disease etiology.

Análisis de antígenos de nematodos parásitos mediante el empleo de anticuerpos monoclonales / Analysis of the parasites nemotodal antigen by means of monoclonal antibody

En el presente trabajo se analizaron extractos de antígenos empleando anti-cuerpos monoclonales, producidos por O. volvulus. La búsqueda de antígeno se realizó en muestras de suero y leche a través de técnicas de ELISA. Esto reveló que la enzima es inmunogénica en hospederos naturales (hombre) y experimentales (ratón), estimulando la aparición de anticuerpos contra la enzima no dirigidos a su centro activo

The vectoral role of several blackfly species (Diptera: Simuliidae) in relation to human onchocerciasis in the Sierra Parima and Upper Orinoco regions of Venezuela.

Experimental and natural infections with Onchocerca volvulus were studied in several anthropophilic blackfly species present in the endemic area of the Upper Orinoco region of Venezuela. When fed on four different Yanomami volunteers in the Sierra Parima, the total infection rate was 31.4% for Simulium guianense in comparison with 7.5% for S. limbatum. The proportion of ingested microfilariae that entered the thorax during the first 24 hours and completed development to the infective stage was also much lower in the latter than in the former species. There was no larval development of O. volvulus in S. antillarum, but one female harboured in its head six infective larvae of a zoonotic filaria. When S. oyapokense s.l. was fed on an infected volunteer in the Upper Orinoco most flies ingested few microfilariae, and further development reached only the L1 stage. Natural infection and infectivity rates for S. guianense in two localities of the Sierra Parima were 0.2-4.0% and 1.3-10.2%, respectively, more than 50% of the L3 larvae being in the head. The corresponding data for S. oyapockense s.l. in the Upper Orinoco were 0.1-0.5%, although the only L3 larva found did not correspond to O. volvulus. It is suggested that S. guianense is the main vector of onchocerciasis in the Sierra Parima and that S. limbatum could play a secondary role. Simulium oyapockense s.l. replaces them in the Upper Orinoco and may maintain a degree of transmission, but its epidemiological importance remains to be assessed.

Morphological differences between Venezuelan and African microfilariae of Onchocerca volvulus.

Comparative morphological and biometric characteristics of microfilariae of Onchocerca gutturosa and O. volvulus from different geographical areas (Upper Orinoco, Venezuela; Togo; Liberia) were assessed. "Stepwise" discriminant analysis and Mahalanobis estimators were applied to measure distance between populations. The results indicate a strong similarity between the two strains from the Upper Orinoco (Venezuela) and the Togo strain, as well as a clear separation between these strains and that of O. gutturosa. The Liberian strain was easily distinguishable from microfilariae from Togo and Venezuela. Discriminant analysis showed the Liberian deme to be as different from the Venezuelan and Togo demes as these demes were from microfilariae of the reference species, O. gutturosa. Although it is necessary to confirm these data using formalin-fixed specimens obtained from the skin, the present findings suggest the existence of geographically-different strains of O. volvulus in America and Africa.

Parasite antigens are present in breast milk of women infected with Onchocerca volvulus.

We used a noncompetitive two-site ELISA with 5 monoclonal antibodies to determine whether parasite antigens are present in breast milk from women infected with Onchocerca volvulus. Seven out of 13 available milk samples contained significant amounts of filarial antigens. Antigen indices in milk correlated with levels of microfilarodermia (Rs = 0.74, P less than 0.005). Antigen-containing milk samples markedly inhibited mitogen-induced proliferation of human mononuclear cells and activated cells within this population that suppressed the proliferative response of autologous lymphocytes to mitogens and antigens. These findings indicate that parasite products are present in breast milk of O. volvulus-infected women and suggest that these may induce immune tolerance and/or suppression in infants born of infected mothers.

Studies on a filarial antigen with collagenase activity.

We examined the ability of two filarial species, Onchocerca volvulus and Brugia malayi, to solubilize collagen molecules from native collagen fibrils. Collagenolytic activity was detected in extracts of adult worms, in living microfilariae of O. volvulus and in live infective larvae and adult female worms of B. malayi. Excretion-secretion factors produced in vitro by infective larvae of B. malayi also contained large amounts of collagenase. Studies with enzyme inhibitors suggest that the latter may be a metallo-protease. Antibodies to filarial collagenase were present in sera from patients with onchocerciasis and brugian filariasis and from mice immunized with B. malayi. These antibodies and a monoclonal antibody raised against O. volvulus antigens immunoprecipitate filarial collagenase but appear not to be directed against the active site of the enzyme.

Experimental and natural infection of Simulium sanchezi by Mansonella ozzardi in the Middle Orinoco region of Venezuela.

Experimental and natural infections of Simulium sanchezi by Mansonella ozzardi were studied in the area of Síquita, Territorio Federal Amazonas, Venezuela. The microfilariae developed synchronously in the blackflies, reaching stage L3 in seven to eight days at temperatures between 23 degrees and 27 degrees C. Larvae in different stages of development, including infective forms, were found in 0.6% of 662 unfed wild-caught females. These results confirm that simuliids are the main vectors of M. ozzardi in the American continent.

Onchocerciasis in Venezuela: prevalence of microfilaraemia in Amerindians and morphological characteristics of the microfilariae from the Upper Orinoco Focus.

Data are presented on microfilaraemia in 191 Yanomami Amerindians from two areas of Venezuela's Upper Orinoco Basin, where an endemic focus of onchocerciasis has been recently detected. Onchocerca microfilariae were found in the blood of 12.6% of the persons examined. In the village with the higher number of examined individuals (N = 162), the prevalence of microfilaraemia in the age groups covaried with the prevalence of microfilariae in the skin and with the parasite load (mf/mg of skin). A positive correlation was found between age and these three variables. The prevalence of Onchocerca microfilaraemia in the Upper Orinoco focus is high, and may be related to the ability of the strain to invade the blood stream. Morphobiometric characteristics of the Onchocerca microfilariae isolated from human blood are similar to those of O. volvulus fixed in formalin from skin and from the uterus of female O. volvulus worms. All have a very short cephalic space. Comparisons with the entity from the Upper Caura river designated as Microfilaria bolivarensis were also made. It was concluded that there are no valid biometric differences in thick blood smears between microfilaria bolivarensis and microfilaria of O. volvulus from the Upper Orinoco deme. A more striking difference may be in the concentration of microfilariae observed in the blood, which in one case of mf. bolivarensis was more than forty times the highest number recorded for microfilaria O. volvulus in Parima.

Development of Onchocerca volvulus larvae in Simulium pintoi in the Amazonas region of Venezuela.

The intake and development of Onchocerca volvulus in Simulium pintoi from the Parima mountain region of the Federal Territory of Amazonas in Venezuela, were studied experimentally. When wild females fed on the lower half of the legs and lower third of the back of an onchocerciasis patient harboring 23 and 264 microfilariae per skin snip, respectively, at each site, an average (median) of 14 (range, 1-77) and 245 (range, 58-495) microfilariae was ingested. However, within 24 hours of microfilarial ingestion a mortality of 47% (16/34 flies) was observed in the group of flies which fed on the back, as compared with 2% (2/101 flies) in the other group which fed on the legs. At a temperature varying between 16 degrees C and 24 degrees C, the development of O. volvulus larvae in S. pintoi was synchronous and orderly; no abnormal nor deformed larvae were observed. Third-stage larvae were first seen in the head of flies dying between 8 and 9 days after microfilarial ingestion, and 98 of 100 larvae recovered from days 10-16 were in the third stage. The proportions of females harboring third-stage larvae among flies which lived through day 8 in the two groups which fed on the legs and back, respectively, were 55% (21/38 flies) and 63% (5/8 flies). Although only two of five positive flies in the latter group contained third-stage larvae in the head (1 and 12, respectively), 71% (15/21 positive flies) of the former group had an average of 2.7 third-stage larvae in the head (range, 1-10).(ABSTRACT TRUNCATED AT 250 WORDS)

Acid phosphatase patterns in microfilariae of Onchocerca volvulus s.l. from the Upper Orinoco Basin, Venezuela.

The patterns of acid phosphatase in strains of Onchocerca volvulus s.l. which parasitize an Amerindian population (Yanomami) in Venezuela's Upper Orinoco Basin were examined by using the naphthol AS-TR phosphate method. The study sample consisted of 40 Yanomami inhabiting a savannah area at 950 m above sea level and 21 Yanomami residents of a tropical rainforest area at an altitude of 250 m. Stained intrauterine microfilariae, still within the egg case, exhibited a diffuse distribution of the enzyme in the early stages of embryonic development and a negative reaction at a more developed stage. Four of the five enzyme staining patterns described by Omar (1978) were found in the 3157 microfilariae examined from skin snips. Their distribution was: Type I--17.2%, Type III--0.5%, Type IV--75.6% and Type V--6.6%. No examples of Type II were observed. The results indicate that acid phosphatase patterns of the Upper Orinoco Onchocerca strain most resemble those of strains from Guatemala and Yemen, and are different from the African strains found in Upper Volta and Liberia. The relative frequency of acid phosphatase patterns was modified by cryopreservation of microfilariae.

Onchocerca volvulus: detection of circulating antigen by monoclonal antibodies in human onchocerciasis.

A monoclonal antibody of the IgM class recognizing Onchocerca volvulus circulating antigen (COA) was obtained. This monoclonal antibody was used in a radioimmunoprecipitation-PEG assay (RIPEGA) to detect circulating antigen in onchocerciasis patients' sera. COA could be detected in 63 (80%) of the 79 African patient sera tested, and in 126 (76%) of the 164 Indian (Venezuela) sera studied. There was no direct correlation between the presence of COA detected in the patient serum and the level of microfilarodermia. The RIPEGA using this monoclonal antibody detected COA in 91% of children under 10 years old, whereas the microfilarodermia in this group was positive in only 52% of the cases. The specificity of this test is improved compared to the results obtained with polyclonal antibodies. Immunofluorescence studies suggest that the COA might be located in the microfilaria cuticle.

Delayed-type hypersensitivity and immunoglobulin E in American cutaneous leishmaniasis.

Delayed-type hypersensitivity (DTH) and both total and specific immunoglobulin E (IgE) antibody levels were studied during an outbreak of American cutaneous leishmaniasis. Direct correlations were detected between DTH reactivity and either age or the period of evolution of the infection, and a clear association with sex (strongest response in females) was observed. Extremely high, age-dependent, total serum IgE levels were measured in the study group, probably due to concurrent intestinal helminthiasis. A low proportion of the group also had detectable levels of specific anti-Leishmania IgE antibody. Total and specific IgE levels were also sex dependent (lowest in females), and an inverse correlation was found between these levels and DTH responsiveness, possibly reflecting the intervention of regulatory influences of T-lymphocyte activity.

T-cell dysfunction and hyperimmunoglobulinemia E in paracoccidioidomycosis.

Various aspects of T and B cell mediated immunity were investigated in 20 well documented cases of active (10) or inactive (10) paracoccidioidomycosis (Pcm), as well as in 8 healthy individuals living in the endemic area of the disease. The results confirm previous reports that active Pcm produces diverse grades of depression of T cell mediated immunity. Such T cell dysfunction is not associated with a reduction in the number of peripheral E rosette-forming cells, and the immunodepression is reversed by chemotherapy. Sera from Pcm (active or inactive) patients have significantly increased levels of total IgE, but the actual proportion of IgE antibodies against P. brasiliensis was very low (0.4-0.6%). The highest levels of total IgE were found in active patients with disease-related immune depression, suggesting that T cell dysfunction might contribute to the excessive IgE production.