Lectin-Mediated Bacterial Modulation by the Intestinal Nematode Ascaris suum.

Ascariasis is a global health problem for humans and animals. Adult Ascaris nematodes are long-lived in the host intestine where they interact with host cells as well as members of the microbiota resulting in chronic infections. Nematode interactions with host cells and the microbial environment are prominently mediated by parasite-secreted proteins and peptides possessing immunomodulatory and antimicrobial activities. Previously, we discovered the C-type lectin protein AsCTL-42 in the secreted products of adult Ascaris worms. Here we tested recombinant AsCTL-42 for its ability to interact with bacterial and host cells. We found that AsCTL-42 lacks bactericidal activity but neutralized bacterial cells without killing them. Treatment of bacterial cells with AsCTL-42 reduced invasion of intestinal epithelial cells by Salmonella. Furthermore, AsCTL-42 interacted with host myeloid C-type lectin receptors. Thus, AsCTL-42 is a parasite protein involved in the triad relationship between Ascaris, host cells, and the microbiota.

Comparison of the effect of the chosen species of saprotrophic fungi on the development of Toxocara canis and Ascaris suum eggs.

The study aim was to compare the antagonistic interaction between saprotrophic soil fungi and embryonic development of geohelminths Toxocara canis and Ascaris suum. The experimental cultures were fertilized eggs of T.canis and A. suum incubated together with mycelium of strains: Fusarium culmorum, Metarhizium anisopliae,Paecilomyces fumosoroseus, Trichoderma viride and Trichothecium roseum. In the control cultures the eggs of both nematode species were incubated without fungi. The experiment was conducted at temp. 26°C for 60 days. Compared with the control, all of the tested species of fungi significantly extended the embryonic development of both T. canis and A. suum. Most inhibitory effect on the rate of embryonic development of T. canis and A. suum had three fungal species: P. fumosoreus, M. anisopliae and T. viride. Compared with the control, on the 60th day of incubation in the presence of each of the tested fungal species, a larger percentage (p<0.05) of morphological abnormalities was stated in developing embryos of T. canis (49­69%) than in A. suum (15.1­67.7%). Among the examined fungal species, only incubation with P. fumosoroseus resulted in significantly greater (p<0.05) incidence of embryonic malformations(embryopathies) in T. canis, as compared with A. suum. Also the percentage of dead larvae of T. canis in the control and in cultures with fungi (12% and 100%, respectively) was significantly higher in comparison with A. suum (0.5% and 10.3­36%, respectively). The highest percentage of non-viable larvae of A. suum was found in the presence of P.fumosoroseus, and the lowest in the presence of M. anisopliae. Findings may indicate that T. canis eggs are more sensitive to antagonistic interaction of the examined fungal strains than A. suum eggs.

In vitro evaluation of the ovistatic and ovicidal effect of the cosmopolitan filamentous fungi isolated from soil on Ascaris suum eggs.

The ovicidal activity of seven fungal strains: Acremonium alabamense, Alternaria chlamydospora, Cladosporium herbarum, Fusarium solani, Paecilomyces variotii, Paecilomyces viridis and Penicillium verruculosum isolated from urban soil samples from Poland was determined in vitro. The fungal mycelium was co-cultured with Ascaris suum eggs on plates with 2% water-agar for 28 days. Eggs exposed and unexposed (control) to fungal mycelium were observed weekly by light microscopy and the percentage of malformed eggs were determined. The eggs were classified according to following parameters: type 1 - biochemical and physiological effect without morphological damage to the eggshell; type 2 - lytic effect with morphological alteration of the eggshell and embryo; type 3 - lytic effect with morphological alteration of eggshell and embryo with hyphal penetration and internal egg colonization. All examined species of fungi extended embryogenesis, but the retardation of embryonic development was varied and depended on the species. A. alabamense, A. chlamydospora and P. verruculosum exhibited very high inhibitory activity on A. suum egg development. The fungus-exposed eggs revealed morphological alternations in all stages of embryogenesis. Isolates of F. solani, P. variotii and P. viridis showed hyphal penetration and internal colonization of A. suum eggs (type 3 effect). No appressoria were produced and simple hyphal penetrations were most commonly observed. A. alabamense and P. verruculosum demonstrated morphological destruction, with eggshell destruction. The remaining fungi showed type 1 effect. The results demonstrated that examined strains of F. solani, P. variotii and P. viridis may be considered to be potential limiting factors of parasitic geohelminth populations.

Biological interactions between soil saprotrophic fungi and Ascaris suum eggs.

The in vitro effect of saprotrophic soil fungi on the embryonic development of Ascaris suum was evaluated. The fungi tested were Aspergillus fumigatus, Aspergillus terreus, Penicillium citrinum, Penicillium expansum, Fusarium oxysporum and Trichothecium roseum, isolated from children's recreation areas in the city of Lodz (Poland). Each species was co-cultured with A. suum egg suspension (6 × 10(3)eggs/ml) at 25 ± 2°C for 60 days. Each day, 100 eggs were randomly collected and their developmental stage was classified macroscopically. Additionally, at days 4, 7, 14, 28, 42 and 60 of incubation, the viability and the percentage of eggs with morphological altered embryo/larva were determined in each sample. Microscopic examination revealed that exposure of eggs to the mycelium of examined fungi inhibited embryogenesis of A. suum. All control culture eggs reached L2 larval stage after 26 days of incubation, while the experimental cultures did so after 32-51 days, depending on the fungal species. Three species were found to exhibit very high inhibitory activity on A. suum egg development: A. terreus, P. expansum and F. oxysporum. Embryopathies and non-viable embryos/larvae were observed significantly more frequently in the eggs co-cultured with fungal species than in control cultures. The fungus-exposed eggs revealed morphological alternations in the early zygotic cleavage, blastula, gastrula and larval stages. After 60 days of incubation with mycelia of P. expansum, A. terreus and F. oxysporum, the mortality of the larvae reached 55.3-60.3%. P. expansum and F. oxysporum showed hyphal penetration and internal egg colonization of A. suum eggs.

Biological control of Ascaris suum eggs by Pochonia chlamydosporia fungus.

Ascaris suum is a gastrointestinal nematode parasite of swines. The aim of this study was to observe Pochonia chlamydosporia fungus on biological control of A. suum eggs after fungus passage through swines gastrointestinal tract. Eighteen pigs, previously dewormed, were randomly divided into three groups: group 1, treated with the fungus isolate VC4; group 2, treated with the fungus isolate VC1 and group 3 did not receive fungus (control). In the treated groups, each animal received a 9 g single dose of mycelium mass containing P. chlamydosporia (VC1 or VC4). Thereafter, animal fecal samples were collected at the following intervals: 8, 12, 24, 36, 48, 72 and 96 h after treatment beginning and these were poured in Petri dishes containing 2% water-agar culture medium. Then, 1,000 A. suum eggs were poured into each dish and kept in an incubator at 26 °C and in the dark for 30 days. After this period, approximately 100 eggs were removed from each Petri dish and morphologically analyzed under light microscopy following the ovicidal activity parameters. The higher percentage observed for isolated VC4 eggs destruction was 57.5% (36 h) after fungus administration and for isolate VC1 this percentage was 45.8% (24 h and 72 h) (p > 0.01). P. chlamydosporia remained viable after passing through the gastrointestinal tract of swines, maintaining its ability of destroying A. suum eggs.

Ovicidal activity of seven Pochonia chlamydosporia fungal isolates on Ascaris suum eggs.

The ovicidal effect of the nematophagous fungus Pochonia chlamydosporia on eggs of Ascaris suum was tested under laboratory conditions. A. suum eggs were plated on 2% water-agar with seven fungal isolates (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) and control without fungus. After 5, 7, 10, 14, 15 and 21 days of incubation, approximately 100 eggs were removed from the plates and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo showing hyphal penetration and internal egg colonization. The isolates effectively destroyed A. suum eggs and all types of effects were observed during the experiment. There was no variation in ovicidal capacity (type 3 effect) among the isolates (p>0.05) throughout the experiment. After 21 days, isolate 5 showed the highest percentages of type 3 effect (58.33%). The results indicated that P. chlamydosporia (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) can destroy A. suum eggs and is, therefore, a potential biological control agent of nematodes.

In vitro evaluation of the effect of the nematophagous fungi Duddingtonia flagrans, Monacrosporium sinense, and Pochonia chlamydosporia on Ascaris suum eggs.

The in vitro effect of four isolates of the nematophagous fungi Duddingtonia flagrans (AC 001), Monacrosporium sinense (SF 53), and Pochonia chlamydosporia (VC 1 and VC 4) on eggs of Ascaris suum was evaluated. One hundred thousand A. suum eggs were plated on 2% water-agar with the grown isolates and control without fungus. After 7, 14, and 21 days, 100 eggs were removed and classified according to the following parameters: type 1, lytic effect without morphological damage to eggshell; type 2, lytic effect with morphological alteration of embryo and eggshell; and type 3, lytic effect with morphological alteration of embryo and eggshell, besides hyphal penetration and internal egg colonization. P. chlamydosporia showed ovicidal activity (p < 0.01), mainly of the type 3 effect, on A. suum eggs in the studied intervals of 13.3% (isolate VC 1) and 17.3% (isolate VC 4), 13.9% (VC 1) and 17.7% (VC 4), and 19% (VC 1) and 20% (VC4), respectively, at 7, 14, and 21 days. The other fungi showed no type 3 effect. P. chlamydosporia is a potential biological control agent of A. suum eggs.

[The influence of the Metarhizium fungi on the embryonic development of Ascaris suum]. / Wplyw grzybów z rodzaju Metarhizium na rozwój embrionalny Ascaris suum.

PURPOSE: The present study focused on the effect of two fungal species Metarhizium flavoviride and Metarhizium anisopliae on the embryonic development and viability of Ascaris suum larvae. RESULTS: Microscope examination revealed that the presence of the mycelium inhibited the egg development, compared to control. The fungus-exposed eggs featured zygote vacuolisation, irregular blastomere cleavage, and morphological disturbances in the stages of blastula, gastrula, and larva. The embryopathies were significantly more frequent in the eggs incubated with M. flavoviride. Also the mortality of the developed larvae was significantly higher after 60-day culture with M. flavoviride.

[Limitation in the population of parasitic geohelminths by saprotrophic soil fungi and their secretions]. / Badania nad ograniczeniem liczebnosci pasozytniczych geohelmintów za pomoca saprotroficznych grzybów glebowych i ich wydzielin.

INTRODUCTION: The aim of the study was to examine the effect of mould fungi isolated from soil and of two of their metabolites, aflatoxin G1 and ochratoxin A, on the embryonic development of eggs and mortality of larvae of Ascaris suum. An attempt was made to demonstrate synergism between fungi species in the action on embryogenesis of Ascaris suum. MATERIAL AND METHODS: Fungi were isolated from soil and cultured on the standard Czapek-Dox agar medium at 26 degrees C for 60 days. Fertilized eggs of Ascaris suum were incubated in PBS solution with the mycelium of each species or without fungi in the control culture. To demonstrate synergism between fungi species, eggs were cultured with three combinations of two species. Additional cultures were done with aflatoxin G1 and ochratoxin A at concentrations of 0.5, 1, and 2 ppm. RESULTS: Microscopy showed a slower rate of development of eggs from experimental cultures as compared to the control culture. Fungi, as well as mycotoxins, caused vacuolization of the zygote, uneven division of blastomeres, and morphological abnormalities of the embryo. The highest mortality (36%) of invasive larvae of Ascaris suum was caused by Paecylomyces fumosoroseus. The present study revealed that the antagonistic effect of a fungi on the development of Ascaris suum eggs is increased or decreased in the presence of another fungi species. Exposure to aflatoxin G1 and ochratoxin A at concentrations as low as 0.5 ppm leads to a greater percentage of deformed embryos and 100% mortality of larvae.

Bactericidal activity in the pig roundworm Ascaris suum.

A potent, humoral, bactericidal activity against Micrococcus luteus was discovered in pseudocoelomic fluid of the pig roundworm, Ascaris suum. The activity, which was not bacteriolytic, was not due to lysozyme or to a dietary antibiotic. It was not inactivated by exposure to 100 degrees C, to low or high pH, or to ethanol. Dialysis, electrophoresis and agar-diffusion experiments suggested that the main antibacterial activity in the fluid was associated with a basic substance of molecular weight somewhat less than 14,000 Da. Two other Gram-positive organisms, Bacillus megaterium and Staphylococcus aureus, were also killed by the Ascaris fluid, but the Gram-negative Escherichia coli, Proteus vulgaris and Bordetella bronchiseptica were insensitive.

Bacterial flora, a possible source of serotonin in the intestine of adult female Ascaris suum.

The effect of intestinal bacteria on 5-hydroxytryptamine (5-HT, serotonin) level and 5-HT turnover rates in Ascaris suum intestine are presented. Ascaris suum were incubated in media containing antibiotics for 24 hr, and the bacterial flora in the anterior regions of the intestine of A. suum was eliminated. The bacteria were significantly reduced (> 99%) but not eliminated in the middle and posterior segments of the worm. The 5-HT level decreased in the intestine after 24 hr incubation in antibiotics, whereas the 5-HT turnover rate increased (131 ng/mg protein/hr). Two possible sources of 5-HT from the intestine were examined: the intestinal tissue itself and the microflora inhabiting the intestine. The 5-HT level in the microflora was 30% higher (72.6 ng/g) than the intestinal tissue (43.3 ng/g) in control samples (0 hr, no antibiotics). These values decreased significantly after 24 hr incubation in A. suum saline. The 5-HT values decreased to 18.6 ng/g in the presence and 28.6 ng/g in the absence of antibiotics. The 5-HT turnover rate during this time period indicated that as the number of bacteria declined, the 5-HT turnover rate also declined in the microflora, but the 5-HT turnover rate in the intestinal tissues increased. Results from these studies suggest that bacterial 5-HT may be contributing to the 5-HT level in A. suum intestinal tissue.

Fitoterapia de la enterobiosis en escolares del Distrito de Ate-Vitarte. Lima-Perú, 1992 / Entherobiosis phytotherapy in students of Ate-Vitarte. Lima-Perú 1992

Se ha trabajado con una muestra poblacional de 190 niños que estudian en el Centro Educativo no estatal (CENE) "Walt Witman" de la Cooperativa 27 de abril, del distrito de Ate-Vitarte Lima. De los 190 niños estudiados, 105 resultaron positivos a enterobius vermicularis, mediante la técnica de Graham modificada. El objetivo principal es el demostrar la eficacia del tratamiento dosificando almendra de la pepa del zapallo (cucurbita maxima) y de las pepas de la papaya (carica papaya). Comprobando primero "in vitro" frente a ascaris soom, para luego determinar las dosis ideales, para ser administradas a los niños en mención, registrándose que el tratamiento con pepas de papaya es más eficaz y mejor tolerado. Estos productos son de bajo costo y no han producido efectos colaterales o secundarios.