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At least 397 records · Page 22Linked to original sources

The effect of infusions of microfilariae (L1 larvae) of Dirofilaria immitis in dogs.

Four pups were given three intravenous infusions of microfilariae over a 7-month period to determine if radiographic changes could be detected in the lungs while sensitivity to first stage microfilariae was being induced. Mild pathological changes occurred but these could not be detected on any of the radiographs. Radiographic changes described by others and 'Eosinophilic lungs' did not result from the immune response to the first stage larvae.

Animals↗

Successful cryopreservation of Wuchereria bancrofti microfilariae.

A fast freezing technique developed by James (1981) significantly improved the yield of schistosome larvae that could be stored in liquid nitrogen. This was adapted by Ham et al. (1981) to the freezing of microfilariae of Onchocerca spp. with excellent results. They found 70 to 80% of the cryopreserved larvae remained viable using this method and since then Ham (personal communication) has successfully applied the same technique to Brugia malayi microfilariae. The further application to Wuchereria bancrofti is described.

Animals↗

The concentration of Mansonella ozzardi microfilariae in skin capillaries.

The densities of microfilariae of Mansonella ozzardi in different regions of the body of infected individuals were studied. The density in venous blood closely approximated to that found in finger capillaries, but was significantly lower than the densities in capillaries of the scapular region and buttocks. A possible explanation for this concentration of microfilariae is the lack of glomi in these two sites, which consequently have a lower flow of blood than areas where glomi occur and where microfilarial densities are lower. No relationship was found between blood and skin densities in the buttocks and scapular region, probably due to the inadequacy of the skin snip method used for the detection of this species of filaria. It was concluded that the most acceptable detection method for M. ozzardi remains the sampling of finger capillary blood. It could not be decided, due to the paucity of data, whether any relationship exists between skin microfilarial densities and the biting sites of the vectors.

Adolescent↗

Comparison of the sensitivity of different geographical races of Onchocerca volvulus microfilariae to ivermectin: studies in vitro.

This study was designed to provide baseline information on the sensitivity of 4 geographical isolates of Onchocerca volvulus microfilariae (mf) (Ghana forest, Ghana savanna, Cameroon forest and Guatemala) to ivermectin, and to develop an in vitro system with which to examine parasites for the possible development of drug resistance. Drug effects were best visualized in the presence of monkey kidney (LLCMK2) feeder cells in the culture system (MEM medium+20% serum), since mf maintained in the absence of cells declined in condition rapidly. Incubation of Ghana forest mf (+cells) in ivermectin (10(-5)-10(-10) M) caused a decrease in motility index (MI) scores in a concentration-dependent fashion; drug effects could be observed as early as 6 h, but cultures maintained for up to 8 d showed greater differences between control and drug groups with increasing time. All 4 O. volvulus isolates and O. lienalis (bovine) were compared for their response to ivermectin (10(-7) M): O. lienalis mf were significantly more sensitive (78%) reduction in MI scores on day 8) than the O. volvulus isolates (33.4-47.7% reduction). O. volvulus microfilariae ex utero generally displayed lower levels of motility and were slightly less inhibited by ivermectin than were skin mf. The in vitro system described can distinguish between the populations of mf studied on the basis of differing MI responses to ivermectin and, when combined with assays to test the infectivity of mf to blackflies following exposure to drug, will provide methods with which to examine parasites for the possible development of resistance.

Animals↗

Antigenic peptides on the surface of Dirofilaria immitis microfilariae.

Various methods of radioiodination were employed to identify peptides on the surface of Dirofilaria immitis microfilariae. Optimum surface radiolabelling occurred with the lactoperoxidase-catalyzed reaction. Two major peptides of 16 and 14 kDa were labelled by this method. These peptides were soluble in Nonidet P-40, were not glycosylated, and showed no signs of disulfide linkages. These peptides were immunoprecipitated by sera from D. immitis-infected dogs, but not by sera from uninfected dogs or sera from dogs with potentially cross-reactive nematode infections. Analysis of the 14 and 16 kDa peptides by two-dimensional gel electrophoresis revealed that the 16 kDa peptide was a single unit with a pI of 5.25 whereas the 14 kDa band was composed of three individual peptides with pI values ranging from 5.6 to 6.1. Iodination by chloramine T resulted in the same panel of labelled peptides but suffered from poor efficiency of 125I incorporation. The viability of microfilariae labelled by the standard Bolton-Hunter method decreased by 50% following the reaction which resulted in the labelling of a variety of internal components.

Animals↗

Studies on a 14-kilodalton surface protein of Onchocerca microfilariae.

A 14-kDa antigen present on the surface of uterine microfilariae of Onchocerca spp. has been identified using monoclonal antibodies. The antigen was also found in skin microfilariae, but in a masked or cryptic form. A complementary DNA clone encoding the epitope recognised by one of the monoclonal antibodies was identified in a lambda gt11 library. Nucleotide sequencing revealed that the 233-bp cDNA fragment codes for the carboxy-terminus of the antigen. The deduced amino acid sequence consists of three hydrophobic domains with high potential for beta-sheet formation. The amino-terminal hydrophobic domain is followed by 4 positively charged residues (positions 22-25) which contribute to the rather basic character of the protein. Another interesting feature of the polypeptide is its richness in phenylalanine (12.7%). From the sequence information, a synthetic peptide was synthesised which was recognised by one of the monoclonal antibodies directed against the 14-kDa antigen and a small number of sera from patients with onchocerciasis. The relevance of this to vaccination is discussed.

Amino Acid Sequence↗

The gene coding for the major sheath protein of Litomosoides carinii microfilariae, gp22, is transcribed in oocytes and embryonic cells.

The transcription and translation of the gene encoding gp22, a major constituent of the microfilarial sheath of the filarial parasite Litomosoides carinii were studied by in situ hybridisation and immunohistology. Transcription of the gp22 gene is confined to oocytes and embryos in the reproductive organs of adult female worms. It starts in oocytes in the rhachis zone, is maximal in multicellular embryos and decreases slowly as the microfilariae develop. Blood microfilariae lack the gp22 transcript. The gp22 gene product is first detectable in parasites recovered on day 32 post infection. Expression of gp22 begins in multicellular embryos in the uteri of mature female worms and can be detected in all further developed intrauterine stages. The gp22 gene product appears to be exported by the embryonic cells and becomes integrated into the sheath where it may contribute to the flexibility of the latter structure.

Amino Acid Sequence↗

Comparative efficacy of moxidectin 2% equine oral gel and ivermectin 2% equine oral paste against Onchocerca cervicalis (Railliet and Henry, 1910) microfilariae in horses with naturally acquired infections in Formosa (Argentina).

On the basis of positive skin snips for Onchocerca cervicalis microfilariae (MF), 45 horses were chosen from 48 in a total of 257 screened on 12 locations in the northeast Province of Formosa (Argentina), and randomly assigned to two treatment groups of 20 horses each, and a nontreated control group of five horses. On Day 14 post-treatment (PT), skin snip samples in the ivermectin-treated (0.2 mg/kg) group were negative for normal viable microfilariae (MF), while horses in the control group maintained their pretreatment level of infection. On the same Day in the moxidectin-treated (0.4 mg/kg) group, 18 horses were negative for MF, but the remaining two had a total of 1 and 2 MF, respectively (equivalent to 10 and 20 MF/g of skin), but all three parasites showed marked cuticular and structural damage. Both horses were negative in a repeat biopsy on Day 21. From Day 3 PT, one ivermectin-treated horse (5%) evidenced an approximate 15 x 2 x 3 cm-sized, apparently nonpainful, oedematous swelling on the ventral midline, 20 cm in front of the navel, which remained unchanged on Day 14 PT. Adverse reactions were not observed in the moxidectin-treated group. Parasitaemia was found in 18.7% of sampled horses (48 of 257), and the number of MF varied between 10-1820/g of skin snip (mean 172). Similar prevalence and total counts had been described previously in 1985 and 1986 in cattle-farm horses in the same area of Argentina; in surveys in Texas (1974) and Louisiana (1995) in the USA, infection rates were also similar, but total counts much higher. It is concluded that moxidectin 2% equine oral gel and ivermectin 2% equine oral paste, were equally 100% effective in the control of O. cervicalis MF. Contrary to ivermectin, moxidectin did not cause post-treatment dermal reactions.

Administration, Oral↗

Surface carbohydrate changes on Onchocerca lienalis larvae as they develop from microfilariae to the infective third-stage in Simulium ornatum.

Use was made of seven FITC labelled lectins as tools to investigate the surface of Onchocerca lienalis larvae as they develop through to the infective third-stage in a natural vector, Simulium ornatum. The lectins were derived from Canavalia ensiformis (Con A), Lens culinaris (lentil), Triticum vulgaris (wheat germ), Arachis hypogaea (peanut), Helix pomatia, Phaseolus vulgaris (kidney bean) and Tetragonolobus purpureus (asparagus pea). Between 70 and 100 living parasites were examined for each developmental stage; i.e. skin microfilariae, late first-stages, second-stages, preinfective third-stages and infective third-stages isolated from the mouth parts of the flies. None of the lectins used bound to the surface of the microfilariae. However, progressive binding to the cuticle of the first- and second-stages was observed using Con. A, lentil lectin and wheat germ agglutinin (WGA). Following moulting to the third-stage, binding of these three lectins declined. Furthermore, as these lectins decreased, peanut and Helix pomatia lectins progressively increased in their binding, despite the fact that they showed little or no binding to the first- and second-stages; stages at which Con A, lentil and WGA were at their maximum. Asparagus pea and kidney bean lectins failed completely to bind to any of the larvae examined. Carbohydrate inhibition tests showed that the lectin was indeed binding specifically to glycoconjugates on the parasite surface. WGA binding was not inhibited by prior incubation with N-acetyl-D-glucosamine, even at high concentrations, but neuraminic acid did completely inhibit its binding. Judging from the patterns of binding on the nematodes themselves, the carbohydrates may not be vector in origin, but derive from the worms. The lectin specificities indicate that initially mannose/glucose type derivatives are present on the surface. Following moulting to the third-stage these are progressively replaced, or overlaid with galactosamine type derivatives, also present on the infective third-stage as it enters the bovine host. The availability of these surface glycoconjugates to attack mediated by natural insect lectins may be of importance in the parasite regulatory mechanisms of the blackfly. Variability in these surface carbohydrates, and in the response to them could well be a contributing factor in the cytospecific variation in S. damnosum susceptibility to geographical variants of O. volvulus.

Animals↗

A probable case of vertical transmission of Onchocerca volvulus microfilariae.

A probable case of vertical transmission of Onchocerca volvulus microfilariae in Dass Bauchi State, Nigeria is presented. A 28-week-old male child delivered by a 39-year-old Fulani woman suffering from onchocerciasis was positive for O. volvulus microfilariae based on skin snip examinations. The child had general body pruritic rash and intermittent fever. Although the child's skin snip on re-examination five weeks later was also positive, the fever had gradually subsided.

Adult↗

Isolation of microfilariae and eggs of Setaria digitata.

A simple procedure of gel-filtration was used to purify microfilariae of the cattle parasite, Setaria digitata from its eggs. The eggs of S. digitata were also isolated from the female worms by picking out the uterine canal. Gel-filtration gave a recovery of 70% for microfilariae.

Animals↗

Eosinophils are the major effector cells of immunity to microfilariae in a mouse model of onchocerciasis.

Mice inoculated with microfilariae of the filarial nematode Onchocerca lienalis clear their parasites over a period of 3-4 months and are highly resistant to re-infection. We have investigated the comparative roles of the eosinophil, macrophage and neutrophil in effecting this parasite clearance, employing agents specifically to perturb cell function in vivo. Using the anti-IL-5 monoclonal antibody TRFK-5, we show that eosinophils are of primary importance in effecting resistance to re-infection. Ablation of macrophages (with carbon) and neutrophils (with the monoclonal antibody NIMP-R14) had no effect on parasite clearance following re-infection. Neutralization of these 3 cell types during a primary infection showed that while the removal of both eosinophils and macrophages caused a small but significant delay in parasite clearance, the depletion of neutrophils had no effect. This report describes the first direct evidence for eosinophil-mediated killing of microfilariae in vivo, and is consistent with Th-2 cell responses previously described in this model.

Animals↗

The fate of ingested Brugia pahangi microfilariae in susceptible and refractory strains of Culex pipiens and Aedes aegypti.

The uptake and migration of microfilariae of Brugia pahangi in susceptible and refractory Culex pipiens and Aedes aegypti were compared after membrane-feeding the mosquitoes on infected blood and anti-coagulant. There was no difference between the strains within each species in the pattern of migration, but in C pipiens only 24% of the microfilariae reached the thorax whereas in A. aegypti 73-6% were successful. In both susceptible strains the filariae in the thorax developed normally to maturity, but in both refractory strains almost all filariae died within a few days.

Aedes↗

The exsheathment and migration of Brugia pahangi microfilariae in mosquitoes of the Aedes scutellaris species complex.

Studies on the Aedes scutellaris complex of mosquitoes showed that refractory and susceptible species can be distinguished by reference to the proportion of Brugia pahangi microfilariae which reach the thorax within three hours of a feed. Migration ceased approximately 1 1/2 hours after feeding in refractory species, in which only 50% of the ingested worms reached the thorax, In susceptible species over 95% of the ingested microfilariae reached the thorax, and migration lasted for three hours after a feed. Differentiation could be achieved by studying the levels of exsheathment, but it is suggested that analysis of migration levels forms a reliable, accurate and quick method of distinguishing refractory from susceptible species.

Aedes↗

Differences in sensitivity of Schistosoma mansoni schistosomula, Dirofilaria immitis microfilariae, and Nematospiroides dubius third-stage larvae to damage by the polyamine oxidase-polyamine system.

The effect of the polyamine oxidase (PAO)-polyamine system on some helminths was examined in vitro. Both Schistosoma mansoni schistosomula and Dirofilaria immitis microfilariae were highly sensitive to this system, the latter more so than the former. In contrast, exsheathed third-stage larvae of Nematospiroides dubius were resistant to the effects of the PAO-polyamine system. After incubation of microfilariae with either spermine or spermidine in the presence of serum containing PAO (bovine serum or human retroplacental serum) or partially purified PAO, damage of worms occurred, compatible with our criteria for worm death. Similar results were obtained with schistosomula by using spermine. The damage seemed to be mediated by PAO products other than hydrogen peroxide because catalase did not protect either parasite. Our data demonstrate that helminths may be damaged by products of the PAO-polyamine system.

Acrolein↗

Canine filariosis caused by Dirofilaria immitis in Mozambique: a small survey based on the identification of microfilariae.

Dirofilaria immitis was diagnosed in 4 of 13 indigenous dogs from the Province of Zambézia, Mozambique, by acid phosphatase staining of microfilariae. The finding reconfirms the occurrence of the parasite in Mozambique after 3 decades and emphasises the need for extensive surveys. Additionally, in 1 of the infected dogs, microfilariae of Dipetalonema reconditum were detected, which is the 1st record of this parasite in Mozambique.

Acid Phosphatase↗

Diethylcarbamazine-mediated clearance of Brugia pahangi microfilariae in immunodeficient nude mice.

Congenitally athymic nude C3H/HeN mice, microfilaremic with Brugia pahangi, were treated with diethylcarbamazine citrate (DEC). A single oral dose (100 mg/Kg body weight) of DEC resulted in the rapid reduction of numbers of circulating microfilariae in nude, thymus-grafted nude and complement-depleted nude mice. Antibodies of the IgM and IgG isotypes were not detected in the serum of microfilaremic nudes or on the microfilarial surface. These results suggest that DEC-mediated clearance of microfilariae from the circulation of nude mice is probably independent of thymus-dependent immunological mechanisms.

Animals↗

Uptake and development of Wuchereria bancrofti in Culex quinquefasciatus that fed on Haitian carriers with different microfilaria densities.

In studies conducted between 1984 and 1986, the vector competency of Culex quinquefasciatus was assessed after bloodfeeding on 61 Haitian volunteers with different densities of Wuchereria bancrofti microfilariae (mf) and on 11 that were amicrofilaremic. Infected volunteers included persons previously given diethylcarbamazine citrate for 12 consecutive days and some that were untreated. Mosquitoes, derived from field-collected larvae, were released under bed nets and fed upon the volunteers while they slept. The mean mf uptake in mosquitoes that fed on 21 carriers with low to ultralow densities (1-28 mf/ml blood) was 0.2-4 mf/mosquito (mean = 1.7 mf). The observed infectivity rate of greater than 3,000 mosquitoes that fed on high (56-7,500 mf/ml blood; median = 525), low (11-49 mf/ml blood; median = 20), or ultralow (1-10 mf/ml blood; median = 3) density carriers was 4, 11, and 30 times higher, respectively, than the expected rate calculated from the estimated volume of the imbibed bloodmeal. These results indicate that "hidden" carriers (less than 50 mf/ml blood) may serve as a source of infection for mosquitoes, and support the increasing evidence that mosquitoes ingest more microfilariae than expected.

Animals↗