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.

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.

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.

Studies on plating efficiency and estimation of viability of suspensions of Paracoccidioides brasiliensis yeast cells.

Mild sonication was used to obtain single cell suspensions of Paracoccidioides brasiliensis. These cells were intact by microscopic criteria. Direct cell counts in a given inoculum and colony formation on various media were used to determine plating efficiency. Sonicated and nonsonicated cell suspensions were used to study plating efficiency and to estimated viability by means of vital dyes. Methylene blue Erythrosin B, and Janus green were unreliable when used with P. brasiliensis, but vital dyes were accurate when tested with Candida albicans. Acridine orange gave more meaningful results of viability. Estimates of viability, however, changed significantly as a result of relatively minor alterations in the composition of the suspending medium. In initial experiments, the plating efficiency of P. brasiliensis was dismally low. It descended abruptly with increasing dilution of inoculum. Efficiency was much improved if horse serum was added to brain heart infusion plates or if glucose glycine yeast extract (GGY) plates were incubated at room temperature and mycelial colonies were counted. With the technique we report, current plating efficiency of sonicated suspensions is of the order of 25%. Our results and procedures have an important bearing upon those studies concerned with in vitro killing of P. brasiliensis in suspensions or with isolating this fungus from clinical or environmental specimens.

New method for estimating digestion of Paracoccidioides brasiliensis by phagocytic cells in vitro.

We describe a method by which phagocytosis and digestion of Paracoccidioides brasiliensis yeast cells by polymorphonuclear leukocytes or other phagocytic cells may be estimated. Suspensions of P. brasiliensis in its yeastlike phase were sonicated, counted, and incubated with known numbers of peripheral blood polymorphonuclear leukocytes. At given intervals, cytocentrifuge droplets were stained by a variation of Papanicolaou's method. Stained preparations were examined with phase-contrast optics. Digested organisms showed total or partial disappearance of protoplasm. Green-stained cell walls resisted digestion. The proportion of digested cells as a function of time was estimated.

Identification and purification of the specific antigen of Paracoccidioides brasiliensis responsible for immunoelectrophoretic band E.

A new purified antigen (E2) of Paracoccidioides brasiliensis mycelial growth phase was isolated by immunoadsorption from a crude metabolic soluble extract of the fungus. The antiserum prepared in a rabbit by inoculation of E2 antigen developed only one immunodiffusion line with the crude metabolic extract. Findings on immunological analysis showed that E2 antigen is the antigenic component of immunoelectrophoretic band E. The isolated antigens did not possess detectable alkaline phosphatase activity. It reacted in immunodiffusion tests with all the sera (14/14) from P. brasiliensis infected patients containing precipitating antibodies.

Isolation of a specific antigen with alkaline phosphatase activity from soluble extracts of Paracoccidioides brasiliensis.

A specific antigen of Paracoccidioides brasiliensis was isolated from a metabolic extract of the fungus. The extraction was made by specific adsorption to and subsequent elution from a column containing a cross linked polymer to which the antibodies of a monospecific rabbit serum had been covalently attached. The purity of the final product was demonstrated by immunodiffusion analysis of the eluate using immune serum produced in a sensitized rabbit. The purified antigen was shown to have cationic electrophoretic mobility and alkaline phosphatase activity.