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At least 217 records · Page 12Linked to original sources

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↗

[The epidemiological significance of neo-natal parasitism with microfilariae of Onchocerca volvulus (author's transl)].

Between April 1977 and June 1978, 214 babies born of 210 mothers infected with Onchocerciasis, were examined at the maternity of Po Hospital (Upper Volta). In four of them (1.9%) dermal microfilariae of Onchocerca volvulus were found during the first week after birth. In two other cases microfilariae were seen in the tissue of the umbilical cord. All specimens from placentas and amniotic fluid gave negative results. The infected babies were reexamined monthly monthly from 2 to 18 months, depending on the cases. One child remained positive until the 6th month follow-up. The base-line examination of 5.757 children less than 2 years old from different villages in the WHO Onchocerciasis Control Programme in the Volta River Basin Area showed that 1% of children less than one year old and 2% of children of 12 to 23 months old were carriers of dermal microfilariae. These results indicate that in meso- or hyperendemic areas children less than one year old and carriers of microfilariae of Onchocerca volbulus may have been infected in utero. The existence of such cases, in an area of vector control, does not prove a residual vector borne transmission.

Burkina Faso↗

Effect of immune serum on Brugia malayi microfilaria: ultra structural observations.

The in vitro effect of immune monkey serum on microfilariae of Brugia malayi was investigated using the electron microscope. The sheaths of microfilaria incubated with immune serum were seen to be covered with electron dense precipitates. After 24 hours of incubation, majority of microfilariae incubated with immune serum exhibited degenerative changes. Comparable changes were not seen in microfilariae incubated with normal serum. The significance of these observations are discussed.

Animals↗

[Correlation between the number of microfilaria ingested and the thickness of the peritrophic membrane of the vector in onchocercosis of the African savannah].

The fact, as previously established, that a great number of microfilariae remain trapped in the peritrophic membrane of S. damnosum, explains the low proportion of microfilariae passing into the hemocoel. However, it doesn't account for the strong "limitation phenomenon" observed in savannah. Histological analysis shows a growth of the peritrophic membrane proportionate to the number of ingested microfilariae. The microfilariae, exciting the digestive epithelium, provoke the liberation of greater quantities of peritrophic substance, thus reinforcing the peritrophic barrier.

Animals↗

Lethal damage to Simulium metallicum following high intakes of Onchocerca volvulus microfilariae in Guatemala.

High intakes of microfilariae of a Guatemalan strain of Onchocerca volvulus by Simulium metallicum proved lethal to a considerable proportion of the flies within 24 hours of taking an infecting blood-meal. The gross histopathological changes resulting from infection were compared in groups of moribund and of apparently healthy flies during this period. The marked morbidity of the flies could be related to certain events occurring in the alimentary tract and haemocoele of the fly soon after ingestion of the parasitized blood. The rapid death of the flies may be ascribed to one or more of the following reasons: (i) reverse migration of numerous microfilariae from the expanded- to the tubular-part of the mid-gut, where they cause serious injury and disintegration of the gut epithelium; (ii) abrasive damage to the stomach epithelial cells by the invading microfilariae with occasional release of the gut contents into the haemocoele; (iii) interruption of the formation of the peritrophic membrane, particularly at its anterior and posterior ends, with subsequent failure of the flies to digest the blood in the stomach; (iv) passage of large amounts of parasitized blood from the stomach backwards into the hind-gut, leading to its mechanical obstruction and (v) invasion and injury of various organs of the fly, among them the ventral nerve-cord, brain, optic nerve, eye, halteres, fat-body and flight musculature by excessive numbers of microfilariae.

Animals↗

Brugia malayi: acquired resistance to microfilariae in BALB/c mice correlates with local Th2 responses.

The role of specific Th subsets in the regulation of acquired resistance to the filarial parasite Brugia malayi is not known. We examined pathologic and cytokine responses in filarial antigen-sensitized BALB/c mice inoculated intraperitoneally with live microfilariae. Animals immunized three times with soluble microfilarial antigen demonstrated accelerated clearance of live parasites (12 +/- 5% of parasites recovered from the peritoneal cavity 4 days after inoculation vs 57 +/- 6% in controls, P < 0.001). Elimination of microfilariae by immunized mice was associated with local eosinophilia (1.5 x 10(7) eosinophils/ml peritoneal wash fluid compared with 2 x 10(5) eosinophils/ml in unimmunized animals), development of local eosinophil-containing granulomas, and elevated serum IgE levels (7.0 +/- 1.4 vs 2.1 +/- 0.9 micrograms/ml in controls, P < 0.01). CD4+ cells from the site of parasite challenge produced Th2-associated cytokines exclusively (IL-4 and IL-5, not IFN-gamma and IL-2) in response to Brugia antigen, whereas spleen and lymph node cells produced both Th1- and Th2-associated cytokines. Mice immunized a single time with microfilarial antigen did not clear parasites in this time, and peritoneal exudate cells from these animals produced IFN-gamma but not IL-5. These results indicate that acquired resistance to B. malayi microfilariae in mice is associated with induction of a Th2 and not a Th1 response at the site of parasite elimination.

Animals↗

Brugia pahangi: the effects of cecropins on microfilariae in vitro and in Aedes aegypti.

Synthetic cecropins, antibacterial peptides from insect haemolymph, have been tested for their ability to attenuate the motility of microfilariae of the filarial nematode Brugia pahangi in an in vitro assay. Fifty micromolar concentrations of these peptides, equivalent to physiological concentrations in immune-stimulated insects, cause significant attenuation of motility compared with untreated microfilariae. Similar results were obtained with cecropins A and B. This is the lowest concentration for which cecropin has been reported to be active against eukaryote organisms. Antiserum to the cecropin homologue sarcotoxin 1A successfully blocked the observed activity. When the same concentration of cecropin B was coinjected with B. pahangi microfilariae into adult females of the mosquito, Aedes aegypti, a significant reduction in the numbers of developing larvae was observed.

Aedes↗

Structure and function of a family of chitinase isozymes from Brugian microfilariae.

A family of chitinase isozymes, consisting of three proteins from the microfilariae of Brugia pahangi and two previously described chitinases from the microfilariae of B. malayi, has been characterized. The five members of this family display closely related chitin-degrading activities, characterized by strong endo- rather than exochitinase activity. All five proteins have highly conserved sequences at their amino-termini and appear to share a two-domain tertiary structure, as demonstrated by proteolysis of the native molecules. The amino-terminal domain appears to be responsible for the enzymatic activity and retains this activity when cleaved and separated from the remainder of the molecule(s). Glycosylation differences are apparent for the isozymes from the two different Brugian species. No representatives of this family could be detected in the microfilariae of another filarial species, Dirofilaria immitis, which differs in several aspects of its lifestyle from the Brugian filariae.

Amino Acid Sequence↗

Lectin binding studies on adult filariae, intrauterine developing stages and microfilariae of Brugia malayi and Litomosoides carinii.

Sections of macrofilariae of Brugia malayi and Litomosoides carinii revealed binding of the gold-labelled lectins WGA, DBA and PNA. Specificity of binding was controlled by competitive inhibition with the respective sugars. N-acetyl-glucosamine, N-acetylgalactosamine and galactose residues seem to be present in the respective tissues. The lectins were bound preferentially to parts of the reproductive organs and to the fluid contents of their lumina. The results of the chitosan test and binding experiments with WGA-gold conjugate suggest the presence of chitin in the sheath of oocytes or zygotes. Binding of WGA could not be inhibited with 0.5 M N-acetylglucosamine, but only with 10 mM triacetyl chitotriose. In older stages, binding of WGA to the sheath could be inhibited by 0.5 M N-acetylglucosamine. In mature microfilariae, the outer surface of the sheath did not show affinity for WGA, but small amounts were bound to the inner surface. Therefore, the sheath of later developmental stages and microfilariae does not contain chitin but only N-acetylglucosamine residues. The degradation of the chitin content might enable the elongation and flexibility of the sheath of microfilariae.

Animals↗

Setaria cervi: enzymes in microfilariae and in vitro action of antifilarials.

Microfilariae of bovine filarial parasite Setaria cervi are equipped with the enzymes of glycolysis, pentose phosphate and PEP-succinate pathways and thus resemble the adult form in its metabolic pattern. Malate dehydrogenase was the most active enzyme in microfilariae followed by lactic dehydrogenase and fumarase, while phosphoglucoisomerase, PEP-carboxykinase and FDP-aldolase were comparatively less active. The very low ratio of PK/PEPCK in S. cervi microfilariae indicates active fixation of CO2 into PEP to produce oxalacetate. Centperazine and diethylcarbamazine significantly inhibited PEP-carboxykinase, fumarate reductase and succinic dehydrogenase, suggesting that these antifilarials probably exert microfilaricidal action by blocking the PEP-succinate pathway.

Animals↗

[Suppression of parasitemia in rodent filariasis (Litomosoides carinii) by immunization with BCG and microfilaria. I. Intracutaneous inoculation of BCG].

After intracutaneous inoculation of BCG and challenge by subcutaneous injection of infective larvae of Litomosoides carinii, the parasitaemia of the filarial infection in cotton rats remains significantly lower when BCG and larvae are applied in the region of the same popliteal and ileal lymph nodes. However, when the infective larvae are directed to other regional lymph nodes (Ln cubitales and axillares), the depression of microfilaraemia is missed. The worm load (recovery rate) and the expulsion of microfilariae by the adult worms are not influenced by the BCG inoculation. Obviously BCG stimulates the lymphatic tissue unspecifically, and the infective larvae produce the first antigen contact, which is boostered by the microfilariae at the onset of patency. When the intracutaneous BCG inoculation is combined with specific antigen stimulation by simultaneous injection of blood microfilariae in the region of the same lymph nodes, the microfilaraemia of the challenge infection disappears completely or remains extremely low.

Animals↗

[Diaplacental transmission of microfilaria of the species, Brugia pahangi, in the cat].

Eight kittens born of two Brugia pahangi infected cats have been studied for transplacental passed microfilariae. In the peripheral blood microfilariae could not be demonstrated at any time. However, in the lung of a young cat killed two days post partum ca. 30 microfilariae have been found (microfilaremia of the mother 90 mf/20 mm3). Histological studies suggested two possibilities of transplacental passage--by blood and by secretion of the uterus glands.

Animals↗

A direct fluorescence technique for the rapid detection of sheathed microfilariae in blood smears.

Thick and thin blood smears containing microfilariae of Wuchereria bancrofti, Loa loa, Brugia malayi, Brugia pahangi, Brugia patei or Acanthocheilonema viteae were prepared from either cryopreserved blood samples or from freshly collected blood, fixed in methanol and treated with a fluoresceinated lectin wheat germ agglutinin. Sheathed microfilariae of W. bancrofti, L. loa, B. malayi, B. pahangi and B. patei in the blood smears could be easily detected and counted using a fluorescence assay. The unsheathed microfilaria of Acanthocheilonema viteae did not fluoresce. The possibility of adapting this technique, which does not require the use of parasite specific antibody for the sensitive, parasitological detection of filarial infections, is discussed.

Animals↗

Laboratory evaluation of a new technique for counting microfilariae in blood.

The technique involves taking a measured volume of blood (20-40 mm3) from a finger or ear prick on to a conventional 3 X 1 in microscope slide and lysing it with water. Microfilariae could either be counted immediately, whilst still alive, or the smear dried, fixed and stained. A comparison between counts of microfilariae in freshly prepared slides and stained slides showed that few microfilariae were lost in the staining process.

Animals↗

On the longevity and behaviour of microfilariae of Wuchereria bancrofti, Brugia pahangi and Dirofilaria immitis transfused to laboratory rodents.

Microfilariae of Wuchereria bancrofti and Brugia pahangi, which were inoculated into Meriones unguiculatus and Mastomys natalensis respectively, were found in the peripheral blood of the rodents until the ninth day after infection, but no microfilarial periodicity was observed. Microfilariae of W. bancrofti disappeared much faster from the peripheral blood of jirds after reinfection than after the first inoculation. Microfilariae of Dirofilaria immitis inoculated into M. natalensis and laboratory mice were recovered in the peripheral blood until the 17th day after infection and remained infective to mosquitoes. For the first few days the microfilarial periodicity observed in the rodents was similar to that observed in the donor dog, but then reversed.

Animals↗

Improved development of Brugia microfilariae following cryopreservation in liquid nitrogen using a technique suitable for field conditions.

A technique for improved cryopreservation at -196 degrees C of Brugia spp. microfilariae has been developed by modifications of a procedure previously used with Onchocerca spp. A double incubation in ethanediol (ED) solutions, firstly at 37 degrees C in 10% (v/v) ED for 15 min and secondly at 0 degrees C in 40% (v/v) ED for 45 sec followed by plunging into liquid nitrogen, resulted in over 90% of the microfilariae of B. malayi exhibiting normal motility. When used with B. pahangi microfilariae, followed by blood-feeding to Aedes aegypti (Bels New strain) a third-stage larvae yield of 4.04 per fly was obtained, compared with 5.16 in the unfrozen group. This represents an infectivity level of 78% compared with unfrozen controls. The technique also has the benefits of being suitable for field conditions, requiring no sophisticated equipment, merely some ice, ethanediol and liquid nitrogen.

Aedes↗

Discrete transcripts encode multiple chitinase isoforms in Brugian microfilariae.

The blood-borne microfilariae of the Brugian nematodes produce multiple isoforms of chitinase, whose expression is coincident with the onset of microfilarial infectivity for mosquitoes. A single cDNA sequence was previously obtained by screening a Brugia malayi microfilarial cDNA library, yet two chitinase isozymes are readily distinguished in this species. In this paper, we present evidence for the existence of multiple transcripts encoding Brugian microfilarial chitinases. Using primers based on the previously-sequenced cDNA clone, we amplified and sequenced two discrete products from B. malayi microfilarial RNA by RT-PCR. While the shorter fragment was nearly identical to the previously sequenced cDNA, the larger fragment contained an extra copy of a serine/threonine-rich repeat. RNAse protection assays were used to demonstrate that both sequences represent true transcripts, and not PCR artifacts. Using primers based on the B.malayi sequence, two novel sequences were generated by RT-PCR from B. pahangi microfilariae. Homologous and cross-species RNAse protection assays verified that multiple transcripts also encode chitinase isozymes in B. pahangi microfilariae.

Amino Acid Sequence↗

Reduced Loa loa microfilaria count ten to twelve months after a single dose of ivermectin.

The action of a single dose of ivermectin (200 micrograms/kg) on 71 Gabonese patients with Loa loa microfilariae in the peripheral blood, and living in areas highly endemic for loiasis, has been evaluated. Ten to 12 months after treatment, 43 patients (63%) had no circulating microfilaria and the geometric mean peripheral blood microfilaria count had decreased by 88.6% (P < 0.02). Thus, a single annual dose of ivermectin can markedly reduce loiasis transmission.

Adolescent↗