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Surface antigens of Litomosoides carinii microfilariae: agglutinating antibodies react with sheath components of 40 and 120 kilo Dalton molecular mass.

This study was conducted to identify surface antigens of the microfilarial sheath of Litomosoides carinii which are accessible to antibodies. Rabbit antisera were raised against the soluble and insoluble fractions of purified sheaths by extracting them with a buffer containing 2-mercaptoethanol and sodium dodecylsulphate. These sera and rabbit hyperimmune sera directed against homogenates of total microfilariae, mature (i.e. microfilariae liberating) female parasites and excretory-secretory products of adult females were able to agglutinate live and formaldehyde-fixed microfilariae. When the antisera directed against sheath constituents were administered to patently infected Mastomys coucha, the microfilaraemia of these animals was rapidly reduced and remained low for a period of 2-3 weeks. Antibodies specifically binding to the microfilarial surface were immunoaffinity-purified on formaldehyde-fixed microfilariae. The antibodies react with sheath antigens of 40 and 120 kDa molecular mass which are produced by the epithelium of the distal uterus of the mature female, secreted and attached to the surface of the sheaths. A 120 kDa antigen recognized by anti-sheath surface antibodies was also detected in the excretory-secretory products of in vitro-cultured immature female L. carinii from day 30 post-infection onwards. In the excretory-secretory products of mature adult female parasites recovered on day 130 post-infection, this 120 kDa molecule was absent. However, material reacting with the antibody was detected in the stacking gel of SDS-polyacrylamide gels. This finding may indicate that the basic units forming the 120 kDa antigen of immature adults or microfilarial sheath surface antigens occur in a highly polymerized form in the excretory-secretory products of mature female parasites.

Agglutination Tests↗

Formation of basement membrane-like structure terminates the cellular encapsulation of microfilariae in the haemocoel of Anopheles quadrimaculatus.

The encapsulation of microfilariae in the haemocoels of mosquitoes combines both humoral and cellular reactions: the microfilariae are first encased in an acellular layer of melanin, followed by a cellular encapsulation by plasmatocytes. In this study, we demonstrated that cellular encapsulation of Brugia pahangi microfilariae in the haemocoel of the mosquito Anopheles quadrimaculatus was terminated by the formation of a basement membrane-like structure on the outermost surface of the cellular capsule. This structure occurred in the early stage of cellular encapsulation and was evident on the exterior surface of the plasmatocyte, when the active haemocytes were attaching to the already melanized microfilariae. The termination structure appears to be laid down by releasing the vesicle inclusions of haemocytes and has similarities in ultrastructure and cationic colloidal gold staining properties with that of mosquito basement membranes.

Animals↗

Effect of N-acetyl-D-glucosamine on the migration of Brugia pahangi microfilariae into the haemocoel of Aedes aegypti.

Two strains of Aedes aegypti (L.), differing in their susceptibility to Brugia pahangi (Buckley & Edeson), were examined with regard to the effect on the proportion of microfilariae migrating from the mid-gut, of specific carbohydrate supplements in the infecting bloodmeal. N-Acetyl-D-glucosamine (GlcNAc), a sugar also present on the microfilarial sheath, significantly increased the migration rate. This enhancement is greater for the refractory strain of Ae.aegypti. The use of sucrose as a control sugar results in no enhancement of microfilariae migration. It is postulated that the GlcNAc is acting by blocking endogenous gut/peritrophic membrane carbohydrate binding proteins, which would normally inhibit microfilariae migration. Furthermore, there is a significant correlation whereby increasing loads of microfilariae ingested result in decreasing proportions migrating across the mid-gut.

Acetylglucosamine↗

In vitro antibody-mediated macrophage activity on Breinlia macropi microfilariae. II. Ultrastructural and video recording investigations of adherence and cytotoxicity.

Video recordings and ultrastructural studies have revealed an intricate sequence of antibody-mediated cytotoxic activity by quokka peritoneal macrophages on Breinlia macropi microfilariae. The microfilaricidal activity was effected by at least two types of macrophages measuring 17 micrometers and 8 micrometers in diameter respectively. The relatively large macrophages were responsible for trapping, encircling and eventually degrading the highly motile microfilariae in a sequence of events in which the participating macrophages may interdigitate. The smaller macrophages adhered transiently to a number of adjacent sites on the surface of a trapped microfilaria, resulting in a series of damaged spots. This activity of the small macrophages was interpreted as responsible for killing the microfilaria. Thus the microfilaricidal activity was interpreted to be accomplished by the co-operative functions of the relatively large and small macrophages.

Animals↗

Injection of microfilariae induces resistance to Brugia malayi infection in ferrets and accelerates development of lymphostatic disease.

Ferrets injected intravenously with living microfilariae or cutaneously with microfilariae followed by intravenous injection developed partial resistance to a challenge infection as measured by recovery of adult filariae from lymphatics. Following a challenge infection, the ferrets injected with microfilariae developed lymphatic pathology characteristic of a chronic infection or that observed following multiple infections. There was disruption of lymphatic drainage of the infected limb and lymphoedema. The results suggest that immune responses to antigens of microfilariae, presumably antigens shared with other developmental stages, effected both increased resistance and enhanced lymphatic pathology.

Animals↗

Microfilaria in a fine needle aspirate of primary solid malignant tumor of the maxillary antrum. A case report.

BACKGROUND: Lymphatic filarial disease is very common in tropical countries like India. The incidental finding of microfilariae in various benign cytologic smears is very common. Microfilariae in malignant effusions have also been reported, but their coexistence with solid malignant tumor at the primary site has not been reported so far. CASE: A 45-year-old male presented with a painful, rapidly growing swelling over the right maxilla that had been present for three months. On routine fine needle aspiration cytology, a microfilaria along with squamous carcinoma cells in a primary solid malignant tumor of the maxillary antrum was found. CONCLUSION: This is the first case reported in English of the incidental finding of a microfilaria in a primary tumor of the maxillary antrum.

Animals↗

Cytochemical localization of carbohydrate residues in microfilariae of Wuchereria bancrofti and Brugia malayi.

We investigated the localization of carbohydrate residues on the surface structures of microfilariae of Wuchereria bancrofti and Brugia malayi, using a panel of 10 different gold-labeled lectins and chitinase. The sheath, a structure that encloses the microfilariae, is not a homogeneous structure, presenting two clearly distinct layers. The outer layer is more electron dense and was not labeled with the lectins. The inner layer is less dense and was intensely labeled with lectins, especially those that recognize D-galactose and N-acetyl-D-galactosamine. Small differences were observed in the lectin labeling pattern of microfilariae of W. bancrofti and B. malayi. D-galactose and fucose were observed in the cuticle of both species. Chitin, as revealed with gold-labeled chitinase, was observed in the cuticle of microfilariae of W. bancrofti but not in B. malayi.

Acetylgalactosamine↗

Comparison of microfilaria concentration method for Setaria digitata infection in cattle and for Dirofilaria immitis infection in dogs.

Several peripheral blood microfilaria concentration methods that use Acetone (Acetone test), 2% formalin (modified Knott method), 5% Tween 20 solution, distilled water, 1% or 0.1% SDS were compared for their efficacy in detecting Setaria digitata microfilaria in cattle. The Acetone test was found to be more efficacious than the modified Knott method or the 5% Tween 20 solution test for detecting the S. digitata microfilaria in bovine blood. However, besides the Acetone test, the modified Knott method was also found to be suitable for Dirofilaria immitis microfilaria detection in dogs. SDS and distilled water were found not to be effective as hemolytic agent for the disruption of the red blood cell of both the cattle and dogs. Thus, the Acetone test is recommended for the primary screening of microfilaremia of S. digitata in cattle.

Acetone↗

Characterization of the high mannose asparagine-linked oligosaccharides synthesized by microfilariae of Dirofilaria immitis.

This report describes the structures of high-mannose-type N-linked oligosaccharides in glycoproteins synthesized by the microfilariae of Dirofilaria immitis. Microfilariae of D. immitis were incubated in vitro in media containing 2-[3H] mannose to allow metabolic radiolabeling of the oligosaccharide moieties of newly synthesized glycoproteins. Glycopeptides were prepared from the radiolabeled glycoproteins by digestion with pronase and fractionation by chromatography on concanavalin A-Sepharose. Thirty eight percent of 2-[3H] mannose incorporated into the microfilariae of D. immitis glycopeptides was recovered in high mannose-type asparagine-linked oligosaccharides which were bound to the immobilized lectin. Upon treatment of 2-[3H] mannose labeled glycopeptides with endo-beta-N-acetylglucosaminidase H, the high mannosetype chains were released and their structures were determined by high performance liquid chromatography and exoglycosidase digestion. The major species of high mannosetype chains synthesized by microfilariae of D. immitis have the composition Man5GlcNAc2, Man6ClcNAc2, Man7GlcNAc2, and Man8GlcNAc2. Structural analyses indicate that these oligosaccharides are similar to high mannose-type chains synthesized by vertebrates.

Animals↗

Microfilariae of Mansonella ozzardi in human skin biopsies.

Microfilariae of Mansonella ozzardi were seen in specimens of skin taken by biopsy from 4 of 6 subjects previously shown to be positive for microfilariae in the blood. While most of the microfilariae were confined to small blood vessels in the papillary and superficial reticular dermis, some were in perivascular spaces and a few were noted in the dermal interstitium. There was no microscopic evidence that the minimal cutaneous disease seen could be directly attributed to the microfilariae.

Adult↗

The BALB/c mouse as a model for immunological studies of microfilariae-induced pulmonary eosinophilia.

Tropical pulmonary eosinophilia (TPE) is believed to result from extreme immediate hypersensitivity to microfilariae localized in the pulmonary vasculature of some persons with lymphatic filariasis. Female BALB/c mice repeatedly immunized by ip injection of Brugia malayi microfilariae become amicrofilaremic within 24 hr of iv parasite challenge, whereas non-sensitized control animals remain patent for greater than 72 hr. Immunized, but not control mice, develop peripheral blood and pulmonary eosinophilia (2,000 cells/mm3 and 65,000 cells/bronchoalveolar lavage, respectively). Serum and bronchoalveolar lavage filarial-specific IgG antibodies are greater in sensitized mice than in controls (ELISA absorbance values 20- and 10-fold higher, respectively). Serum IgE antibody levels are also greater (P less than 0.01) in immunized parasite-challenged mice than in controls (mean cpm 125I-labeled anti-mouse IgE bound to B. malayi antigen-coated Sepharose beads: 7,852 vs. 1,741, respectively). This model exhibits several of the major features of human TPE: amicrofilaremia, elevated levels of serum IgG and IgE antibodies to microfilariae, and blood and pulmonary eosinophilia. This model may be useful in the examination of the role of filarial antigen-specific lymphoid cells and antibodies in regulating the pathologic responses to microfilariae trapped in the lung.

Animals↗

Release of prostaglandin E2 by microfilariae of Wuchereria bancrofti and Brugia malayi.

To elucidate the local release of immunomodulatory prostaglandins by intravascular filarial parasites, the formation of prostaglandin E2 (PGE2) was examined in individual microfilariae of Wuchereria bancrofti and Brugia malayi. Following incubation of living microfilariae immobilized in an agar matrix, prostaglandins released by the parasites were fixed by carbodiimide and localized by indirect immunofluorescence. Prostaglandin E2 was specifically detected around the entire surface of microfilariae with anti-PGE2 antiserum, but not with control nonimmune or PGE2 affinity-immunoadsorbed antiserum. These results provide direct evidence that individual microfilariae of W. bancrofti as well as B. malayi release prostaglandins into their microenvironment. The release of PGE2 by these intravascular parasites may modulate host leukocyte responses, and thereby contribute to the immune defects observed in infected humans with peripheral microfilaremia.

Animals↗

Onchocerciasis in Guatemala. III. Daytime periodicity of microfilariae in skin.

Periodicity of microfilariae of Onchocerca volvulus in the skin was not believed to exist until investigators in Africa recently demonstrated diurnal periodicity. Interestingly, this maximal density of microfilariae in the skin has been shown to coincide with the peak biting time of the Simulium vector. The current study was performed as part of other studies in Guatemala. Maximal density of microfilariae in the skin was at 1000 hours or shortly therafter. This peak is earlier than reported in Africa. The most active feeding period of Simulium ochraceum, believed to be the principal vector in Guatemala, had been previously shown to be from 0800 to 1000 hours. The current report lends support to other findings indicating that Onchocerca volvulus microlilariae have a diurnal periodicity. In addition, the occurrence of peak vector biting time at the same time as maximal numbers of microfilariae in the skin strengthens a hypothesis that these synchronous cycles have a biological significance.

Adolescent↗

[Observation on the susceptibility of Anopheles sinensis to periodic Brugia malayi microfilariae].

AIM: To explore the susceptibility of Anopheles sinensis to Brugia malayi microfilariae of different densities. METHODS: The survival rate of mosquitoes, the rate of microfilariae penetrating through midgut, the mature rate of larvae into infective larvae (L3) and the mean number of L3 in the survived mosquitoes were observed. RESULTS: With the microfilariae densities of 32.5 mf/microliter and 141.5 mf/microliter, the rate of larvae matured into L3 was 36.2% and 8.7%, respectively, the mean number of L3 recovered from infected mosquitoes was 8.24 and 0.30, respectively. CONCLUSION: An apparently higher susceptibility of An. sinensis to B. malayi was found by using B. malayi microfilariae density of 32.5-66.4 mf/microliter.

Animals↗

Surface area calculation for Onchocerca volvulus microfilariae.

The pathogenesis of tissue damage in filarial infections involves the generation of inflammatory mediators by interaction of parasite surfaces with the host immune response. In the presence of a heavy parasite burden, the tremendous potential for development of adverse local or systemic inflammatory responses may be appreciated by knowledge of the cumulative surface area represented by circulating microfilariae. The approximate surface area of a single microfilariae of Onchocerca volvulus was calculated using mathematical formulae for standard geometrical shapes and values derived from serial measurements of electron micrographs. Numerical values for the surface area represented by large numbers of microfilariae are given and compared to surface areas of the host which can harbor many millions of microfilariae.

Animals↗

Emergence of Onchocerca volvulus microfilariae from skin snips before and after treatment of patients with ivermectin.

As ivermectin is supposed to have a paralyzing effect, the emergence of Onchocerca volvulus microfilariae from skin snips before and after treatment was examined. Before treatment 79.8% of the microfilariae emerged from the skin snips. Three days after treatment the microfilarial skin density had dropped to 83% (placebo), 28% (100 microgram/kg ivermectin), 19% (150 micrograms/kg) and 14% (200 micrograms/kg) of the pretreatment values but the microfilarial emergence was not altered distinctly. The majority of the remaining microfilariae were mobile and able to leave the biopsy. Only a slight, but significant, dose-dependent trend of reduced emergence could be shown. From our results no direct support is derived for the hypothesis that O. volvulus microfilariae are paralyzed or immobilized by ivermectin.

Analysis of Variance↗

Chemotherapy of onchocerciasis with high doses of diethylcarbamazine or a single dose of ivermectin: microfilaria levels and side effects.

Fifty adult male subjects with moderate to heavy onchocerciasis from the Liberian rain forest were selected for a double-blind placebo-controlled chemotherapy study. The effects of high doses of diethylcarbamazine (DEC) - 30 mg/kg/d - over one week preceded by a one week initial treatment with normal oral doses of DEC or DEC lotion were compared with a single dose of ivermectin (150 micrograms/kg) and placebo. During the initial treatment DEC tablets or lotion caused distinctly more frequent and severe reactions than did invermectin. The reactions to ivermectin did not differ from those of the placebo patients. High doses of DEC caused, in about half of the patients, headache, dizziness, nausea or vomiting. DEC markedly increased the number of corneal microfilariae and of corneal opacities compared to ivermectin. All changes resolved with a return to pretreatment findings two months after treatment. The three treatment groups showed no differences at the ten months follow-up. In all treated patients skin microfilaria counts fell almost to zero by the end of the two week therapy. In the ivermectin group microfilaria counts remained significantly lower than in the DEC patients at the two and ten months examinations. In summary, ivermectin was much better tolerated than DEC and had a longer lasting effect on the microfilariae in the skin. Since high doses of DEC were less effective and caused more frequent and severe side effects, this approach cannot be recommended for treatment of onchocerciasis.

Adult↗

Cytologic detection of Wuchereria bancrofti microfilariae in urine collected during a routine workup for hematuria.

A 23-year-old Guyanese man experienced intermittent, total, painless, gross hematuria for a month for which he sought medical attention at the Kings County Hospital Center in Brooklyn. Hematuria was accompanied by weakness but not by frequency of urination or burning on urination. Catheterized urine at the time of cystoscopy and each of two subsequent voided specimens examined cytologically contained sheathed microfilariae. Distinguishing features of the microfilariae were well demonstrated with the Papanicolaou stain. The well-stained nuclei, which did not extend into the clear zone, and the distinct, pale-stained sheath led to the positive identification of the microfilariae as Wuchereria bancrofti. The Papanicolaou stain may well be the stain of choice for the identification of microfilariae in the blood. The excellent detail obtained with this routine cytologic stain is as good as that with Giemsa, which does not stain the sheath.

Adult↗