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Large scale production of the vertebrate infective stage (L3) of Onchocerca volvulus (Filarioidea: Onchocercidae).

Laboratory-derived Simulium yahense and S. sanctipauli females were used to produce large numbers of Onchocerca volvulus infective stage larvae (L3) for use in assessing the possible chemoprophylactic potential of ivermectin in the chimpanzee model. Engorgement rates and subsequent post-prandial survival were correlated with the time at which adult flies were offered a carbohydrate source following emergence and the age of flies that fed on microfiladermic volunteers.

Animals↗

Occult filariasis in crows (Corvus brachyrhynchos brachyrhynchos Brehm) infected with Splendidofilaria caperata Hibler, 1964 (Nematoda: Filarioidea).

Adult Splendidofilaria caperata Hibler, 1964 were found in the tunica media of the pulmonary arteries of 21% of 341 crows (Corvus brachyrhynchos brachyrhynchos Brehm) wintering in southern Ontario, Canada. Microfilariae of S. caperata were not observed in the peripheral blood or in skin snips of infected crows. Microfilariae were rarely found in lung blood and then only in small numbers. Microfilariae were apparently overcome in the wall of the pulmonary artery; chronic inflammation was associated with their presence in arterial tissue. Crows apparently acquire S. caperata from insects that hae fed on birds in which a microfilaraemia does develop.

Animals↗

Pathology and epizootiology of Dirofilaria scapiceps (Leidy, 1886) (Nematoda: Filarioidea) in Sylvilagus floridanus (J.A. Allen) and Lepus americanus erxleben.

Dirofilaria scapiceps was found between the synovial sheath and tendons, i.e., within the tendon sheath, in the ankle region of eastern cottontail rabbits (Sylvilagus floridanus) and snowshoe hares (Lepus americanus). In cottontail rabbits, tendons and sheaths appeared normal and all worms were adults. Only one (4%) of 24 infected rabbits contained dead worms. All female worms were gravid in rabbits killed in late winter or early spring. Microfilaremias in rabbits were high (approximately 30-100 microfilariae/60 microliter blood) and of long duration (at least 8-28 mo), and rabbits were considered normal hosts of D. scapiceps. In some snowshoe hares, tendons and sheaths also appeared normal; however, in other hares a chronic proliferative tenosynovitis, characterized by fibrinous exudate, hyperplasia and hypertrophy of the intima and inflammatory cell (predominantly lymphocytes and plasma cells) infiltration of the intimal and fibrous layers of the synovial sheath led to encapsulation of worms. Dead subadult, dead adult, and live adult worms were found in the ankles of hares; 86 (46%) of 186 infected hares contained some or only dead worms. Fibrosis commonly occurred around dead worms. Dead subadults were also found in subcutaneous connective tissues over the trunk of the body. Degenerate embryos and amorphous material were observed in uteri of some female worms in hares killed in late winter or early spring. Few (1-5 microfilariae/60 microliter blood) or no microfilariae were observed in the peripheral blood of hares and microfilaremias were of short duration (less than 8 mo). Microfilariae in hares are probably trapped and destroyed in the chronic inflammatory lesions in the tendon sheaths since normal, degenerate, and calcified microfilariae were observed in the capsules around adult worms. Some microfilariae might also be destroyed in lymph nodes. Although D. scapiceps can be maintained within snowshoe hare populations, hares are considered abnormal hosts of D. scapiceps. Dirofilaria scapiceps may have spread from cottontail rabbits to snowshoe hares relatively recently.

Alberta↗

Antifilarial activity of Mallotus philippensis Lam. on Setaria cervie (Nematoda: Filarioidea) in vitro.

The effect of aqueous and alcoholic extracts of the leaves of Mallotus philippensis (Lam.) Muell. Arg. was studied on the spontaneous movements of the whole worm and nerve-muscle (n.m.) preparation of Setaria cervi and on the survival of microfilariae in vitro. Both the extracts caused inhibition of spontaneous motility of whole worm and the n.m. preparation of S. Cervi characterized by initial stimulation followed by depression in amplitude. The tone and rate of contractions remained visibly unaffected. Aqueous extract at higher concentration showed immediate reduction in tone. The concentration required to inhibit the movements of n.m. preparation was 1/5th for aqueous and 1/11th for alcoholic extract compared to that for the whole worm, suggesting a cuticular permeability barrier. The stimulatory response of acetylcholine was blocked by aqueous extract on whole worm movements. On the microfilariae the LC50 and LC90 were 18 and 20 ng/ml for aqueous and 12 and 15 ng/ml for alcoholic extracts respectively.

Animals↗

[Deviation of the life cycle of Dipetalonema viteae (Filarioidea)].

Dipetalonema viteae is a filarial that can evoluate among hosts zoologically broadly apart (Ixodides and Argasides), but always gathered from meriones burrows. Its evolution is, on the contrary, blocked among most of the other ticks, particularly among Ornithodoros erraticus morphologically very similar to the normal vector. Our work concerns the experimental deviation of the cycle of Dipetalonema viteae, with its possible adaptability in the bosom of an intermediate of fowl tropism and a permanent host, different from a gnawing.

Animals↗

Tetrapetalonema (T.) panamensis (McCoy, 1936) comb. n. (Filarioidea: Onchocercidae) in Colombian primates, with a description of the adults.

Microfilaria panamensis McCoy, 1936 was identified in the blood of Cebus apella, C. capucinus, C. albifrous, and Saguinus oedipus from various regions in Colombia. Adult worms, recovered from the subcutaneous connective tissues of C. apella and S. oedipus, were identified with this microfilaria and are described as Tetrapetalonema (T.) panamensis (McCoy, 1936) comb. n. This nematode is smaller configuration of the female tail and in the appearance of the spicules, and T. barbascalensis which differs in the placement of the vulva relative to the base of the esophagus and in the structure of the right spicule. The spicular ratio of T. panamensis (1.6 to 2.1) is lower than that of any other member of the subgenus (2.6 to 10.8). The host induced and/or geographic variation of microfilariae and adults of T. panamensis is discussed.

Animals↗

Comparative susceptibility of species A, B and C of Anopheles quadrimaculatus complex to infection with subperiodic Brugia malayi and Brugia pahangi (Nematoda: Filarioidea).

Susceptibilities of natural populations of sibling species A, B and C of the Anopheles quadrimaculatus complex and the colonized strain A to subperiodic Brugia malayi and Brugia pahangi were compared. All 3 sibling species showed varying degrees of susceptibility to both B. pahangi and B. malayi, and they were considerably more susceptible to B. pahangi than to B. malayi. The rate and intensity of infection to B. pahangi were highest for species A (66.2% and 7.4 L3/female, respectively) and lowest for species B (21.3% and 1.7 L3/female). For B. malayi these values were higher for species A (29.7% and 1.84 L3/female) than for species B (13.3% and 0.86 L3/female) and C (12.6% and 0.75 L3/female). The colonized strain A of An. quadrimaculatus was significantly more susceptible to both Brugia species than the natural populations of sibling species A, B and C.

Animals↗

[New details on the morphology of Onchocerca lupi (Nematoda, Filarioidea)].

Basing on the fragments of typical specimens new details of morphology of O. lupi females are given. The presence of cuticular thickenings in the form of half rings, with the bifurcated or turned ends is stated. On the surface of cuticula of microfilariae there is very delicate regular transversal striation. The morphometrical features of O. lupi with descriptions of close species were compared.

Animals↗

[Indirect immunofluorescence in filariasis. II. Humoral response in a population living close to a patient with confirmed Wuchereria bancrofti (Nematoda: Filarioidea), Cuba, 1983].

This paper deals with the study by Knott technique, membrane filter and indirect immunofluorescence, of 256 individuals living in Pinar del Rio City, neighbors of a patient suffering elephantiasis produced by Wuchereria bancrofti, 111 donors of the blood bank in the same city, and 10 patients coming from endemic areas who were hospitalized at the "Pedro Kouri" Tropical Medicine Institute, with certainty diagnosis of bancroftiasis. Circulating microfilariae were not found in the neighbors of the positive case, however, a high antifilaria antibody titer (1:512) was observed in the neighbors, suggesting it that in any time this population could have been in contact with this parasite.

Animals↗

The use of cell-conditioned medium for the in vitro culture of Onchocerca spp. larvae (Nematoda: Filarioidea).

The effects of co-culture with monkey kidney cells (LLCMK2), cell-conditioned medium and decreased atmospheric oxygen on the in vitro molting and viability of infective stage larvae (L3) of Onchocerca lienalis and O. volvulus were examined. O. lienalis L3 were cultured in an RPMI 1640-based medium in the presence of an LLCMK2 cell monolayer or in medium which had been conditioned for three days by cells. In paired experiments cell conditioned medium alone in 95% air/5% CO2 produced molting levels of 54 +/- 14% which increased to 67 +/- 20% in treatments cultured under decreased oxygen; this value equalled the level of molting of worms cocultured with LLCMK2 cells. Worm survival in the three environments was similar. In seven additional experiments using O. lienalis (n = 186), overall levels of 74 +/- 12 percent molting and 75 +/- 7% viability on days 21-33 were obtained. Worms increased in length from 503 +/- 50 mu as L3 to 638 +/- 74 mu as L4 on day 21 (p = 0.0001, n = 42-44). Ultrastructural comparison of an in vitro derived L4, (39 days in culture) vs a vector-derived L3 revealed fewer annulations and decreased osmiophilia on the epicuticle of the L4 while the hypodermis showed increased morphogenetic definition. O. volvulus molted at an average rate of 74% (n = 40) with a mean viability on day 28 of 95%. L3 increased in length from a mean of 635 +/- 50 mu to 775 +/- 45 mu as L4 on day 28 (p = 0.0001). Larvae of both species were cultured under these conditions for periods of time exceeding 100 days.

Animals↗

Demonstration of anti-cuticular antibodies by immuno-electron microscopy in sera of mice immunized with cuticular extracts and isolated cuticles of adult Dipetalonema viteae (Filarioidea).

The immunogold technique was used for the ultrastructural localization of antibody-binding sites on thin sections of Lowicryl K4M embedded adult females, infective larvae and pieces of adult cuticles of Dipetalonema viteae insoluble in SDS-2-ME. The antisera used were either produced against SDS-2-ME extracts of cuticles or the insoluble pellet after SDS-2-ME extraction. With both types of antisera a labelling of epitopes on fibers was achieved in intact cuticles. In isolated cuticles the corresponding structures were absent. The same sera crossreacted with the larval and microfilarial cuticle as well as with somatic structures of all three stages. The only serum against isolated cuticle, which did not recognize cuticle fibers also did not crossreact with somatic structures. The recognition of the electron dense cortical layer insoluble in SDS-2-ME depended on the number of immunizations. A labelling of the filarial surface was never achieved. A dense labelling of the apical membrane enfoldings of the hypodermis pointed to an involvement in the synthesis of the nematode cuticle. The rough endoplasmic reticulum in the apex of the epithelial cells of the uterus, the content of the nutrient channels, and the substance between eggshell and microfilariae crossreacted with most of the antisera. This led to the conclusion that these substances are partly produced by the uterus epithelium.

Animals↗

Studies on Dipetalonema viteae (Filarioidea) I. Microfilaraemia in hamsters in relation to worm burden and humoral immune response.

The course of a primary infection with Dipetalonema viteae was studied in one randomly bred and in one inbred strain of hamster. Worm recovery and the duration and intensity of the microfilaraemia were analyzed and related to the humoral immune response of the host by using the indirect immunofluorescent antibody test on frozen sections of female worms, on eggs and on intact microfilariae. The inbred strain showed a greater susceptibility to the parasite. This was evidenced by high worm recovery and prolonged microfilaraemia. The duration of microfilaraemia did not depend on the number of recovered female worms. Most of the randomly bred hamsters suppressed microfilaraemia by week 30 post infection whereas some hamsters of the inbred strain were still microfilaraemic. Splenectomy prior to infection did not affect the duration of microfilaraemia. Antibodies to the cuticle of microfilariae always appeared in the sera after immunity to circulating microfilariae had been built up.

Animals↗

Studies on Dipetalonema vitae (Filarioidea). II. Antibody dependent adhesion of peritoneal exudate cells to microfilariae in vitro.

Peritoneal exudate cells from normal uninfected hamsters adhered in vitro to microfilariae in the presence of 19S antibody fractions from hamsters which had suppressed or were going to suppress their microfilaremia. The adhering cells were predominantly mononuclear, although eosinophils were occasionally found. Experiments with sensitized microfilariae and peritoneal exudate cells indicated that the macrophage probably recognizes the microfilariae/antibody complex. Macrophage cytophilic antibodies did not seem to be involved. This adhesion reaction may initiate the trapping of microfilariae in vivo, thus contributing to the observed acquired immunity to circulating microfilariae in the hamster.

Animals↗

Preliminary data on the genetic differentiation of Onchocerca volvulus in Africa (Nematoda: Filarioidea).

Data are reported on the genetic structure of three Onchocerca volvulus populations, respectively from Mali (savanna), Ivory Coast (forest), and Zaire (forest gallery in savanna). Electrophoretic analysis, carried out on 25 gene-enzyme systems, has shown a remarkable genetic heterogeneity existing within O.volvulus. Zaire and West Africa populations appear chiefly differentiated at Mdh-1 and 6Pgdh loci, their average Nei's genetic distance being 0.11. In West Africa Nei's D found between the savanna and forest samples is 0.04. The savanna population from Zaire is more similar to the savanna one from Mali (D = 0.09) than to the forest one from Ivory Coast (D = 0.13). This appears mainly due to the loci Ldh and Hbdh (possibly linked), some alleles of which seem to be selected for in forest populations (Ldh110, Hbdh108), while others in the savanna ones (Ldh100, Hbdh100). The hypothesis that the discrepant epidemiological patterns of human onchocerciasis are related to intrinsic differences in the parasite seems supported by the obtained data. The differences in allele frequencies found at the reported loci appear strong enough to allow biochemical identification of O. volvulus populations from different geographic regions and different habitats.

Alleles↗

Immunity to Dipetalonema viteae (Filarioidea) infections in resistant and susceptible mice.

The course of infection with Dipetalonema viteae in mice shows marked genetically-determined strain variation. Subcutaneous implantation of 5 female D. viteae into C57BL/10 (B10) mice results in a short term, low level microfilaraemia compared with that seen in similar infections in BALB/c mice. Adult worm survival is similar, thus the different patterns of infections reflect responses directed against the microfilariae larvae (mf). A number of immunological parameters have been monitored during infection in an attempt to identify strain differences which may be correlated with levels of resistance. Blast cell activity in the spleen and lymph nodes showed little strain difference, peaking on day 10 and declining as mf disappeared from the circulation. Total serum IgG levels doubled in both strains during infection, the response being more rapid in B10 mice. Serum IgM levels increased threefold in BALB/c mice but fourteen-fold in B10. Radiosorbent assays identified comparable anti-adult antibody and anti-mf homogenate IgM antibody responses in both strains. Immunofluorescent assay showed that the appearance of IgM antibodies directed against the mf surface correlated with the clearance of mf from the blood of B10 mice, whereas similar antibodies were not detected in BALB/c mice. It is proposed that the efficient clearance of mf in B10 is mediated through an IgM-dependent mechanism and that the chronic microfilariaemia seen in BALB/c mice is facilitated by the absence of a specific IgM response to mf surface antigens.

Animals↗