Joanna Blaszkowska

Geohelminth egg contamination of children's play areas in the city of Lodz (Poland).

The contamination of soil and sand with helminth eggs in childrens play areas in Lodz (Poland) was assessed over two seasons using the flotation method with saturated sodium nitrate solution. A total of 88 samples were examined from 7 childrens playgrounds from various public parks, 6 sandpits situated in school or kindergarten areas and 9 school sports fields. The differences in the number of positive samples from these sites were significant (χ(2)=21.83, d.f.=2 and p<0.0001). The highest rate of contamination was found in the area around sports fields. (15.7%). There was a significant difference between the frequencies of positive samples from the surface and from the deeper layers of the examined sites (χ(2)=11.41, d.f.=1, and p=0.0007). The average density of geohelminth eggs in 100g of soil or sand was 1.1 from sports fields, 0.4 from playgrounds and 0.07 from fenced sandpits. Throughout the study, 4 genera of nematode eggs (Toxocara, Uncinaria/Ancylostoma, Ascaris, Trichuris) and 1 genus (Cystoisospora spp.) of oocysts were detected. A total of 62 eggs were recovered, and 43.5% were fully developed to embryonated egg stages. The contamination rate was different in autumn 2010 and spring 2011, but there was no significant difference in the number of positive findings between these seasons. The helminth eggs were found in 10.9% and 7.6% of samples collected in the spring and in the autumn, respectively. The most frequently seen eggs were from Toxocara sp., which were the most prevalent in both seasons.

Trichomonas vaginalis: pathogenicity and potential role in human reproductive failure

PurposeTrichomonas vaginalis, which colonizes the genitourinary tract of men and women, is a sexually transmitted parasite causing symptomatic or asymptomatic trichomoniasis. The host–parasite relationship is very complex, and clinical symptoms cannot likely be attributed to a single pathogenic effect. Among the many factors responsible for interactions between T. vaginalis and host tissues, contact-dependent and contact-independent mechanisms are important in pathogenicity, as is the immune response.Methods This review focuses on the potential virulence properties of T. vaginalis and its role in female and male infertility.ResultsIt highlights the association between T. vaginalis infection and serious adverse health consequences experienced by women, including infertility, preterm birth and low-birth-weight infants. Long-term clinical observations and results of in vitro experimental studies indicate that in men, trichomoniasis has been also associated with infertility through inflammatory damage to the genitourinary tract or interference with sperm function.ConclusionThese results contribute significantly to improving our knowledge of the role of parasitic virulence factors in the development of infection and its role in human infertility.

Presence of Toxocara spp. eggs in children's recreation areas with varying degrees of access for animals

INTRODUCTION AND OBJECTIVE The contamination, seasonal and vertical distributions of Toxocara eggs in childrens recreation areas were estimated with respect to their accessibility to domestic and stray animals. MATERIALS AND METHODS During autumn 2011 and spring 2012, a total 88 composite samples of soil/sand (300g each) were taken twice, from 2 depths, from 11 sandpits and 11 play areas situated in the city of Łódź, Poland. From the collected material, 528 samples (20g) were tested using the flotation method. Half the sample sites were secured from access to dogs and cats, while the other half were not. RESULTS The difference in the numbers of positive samples from sandpits and playing areas was significant (c 2 = 13.72, p = 0.0002). The highest rate of contamination was observed in poorly-secured play areas (15.8% of positive samples and 1.2 eggs/100 g of soil/sand). The average density of Toxocara eggs in secured play areas was 6 times less than that found in unsecured areas, while secured sandpits were 3 times less contaminated than those unsecured. The contamination rate was similar in autumn 2011 and spring 2012 (6.4% and 6.8%, respectively). An inverse relationship between the sand/soil depth and number of recovered Toxocara eggs was observed. Additionally, other intestinal helminth eggs (Ancylostomidae, Ascaris spp., and Trichuris spp.) and oocysts of Isospora spp. were also detected from soil samples collected from playing fields. CONCLUSIONS The number of Toxocara eggs recovered decreased following fence construction around the examined childrens play areas, but it did not sufficiently prevent the contamination by eggs. These data indicate the necessity for educational programmes which should be implemented for the protection of the local child population from zoonotic infection.

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 childrens 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.

Potentially pathogenic yeasts from soil of children’s recreational areas in the city of Łódź (Poland)

ObjectivesYeasts may become potential human and animal pathogens, particularly for individuals with a depressed immune system. Their presence in the environment, especially in soil, may favour their spread into human ontocenoses.Materials and MethodsEighty-four soil samples obtained from 21 children’s recreational sites in Łódź in autumn 2010 and spring 2011 were evaluated. The yeasts were isolated by classical microbiological methods and identified on the basis of morphological and biochemical features.ResultsThe fungi were found in 73.8% and in 69.0% of the examined samples collected in autumn and spring, respectively. Among 97 isolates of yeasts, the species potentially pathogenic to humans and animals were Candida colliculosa, C. guilliermondii, C. humicola, C. inconspicua, C. lambica, C. lusitaniae, C. pelliculosa, C. tropicalis, Cryptococcus albidus, C. laurentii, C. neoformans, C. terreus, Kloeckera japonica, Geotrichum candidum, G. penicillatum, Rhodotorula mucilaginosa, R. glutinis, Saccharomyces cerevisiae, Sporobolomyces salmonicolor and Trichosporon cutaneum. The most frequently isolated fungi included the genus Cryptococcus (38 isolates) and two species: Rhodotorula glutinis (15), Trichosporon cutaneum (14). C. neoformans, an etiological factor of cryptococcal meningitis, was present in the sandpits of 3 kindergartens. The Candida species were identified from park playgrounds and school sports fields mainly in autumn 2010 (14 isolates), in spring 2011 — only 1 isolate. The concentration of fungal species in particular samples varied considerably, but in the majority of samples, fungi were present at concentration of up to 1×102 CFU/1 g of soil.ConclusionsYeasts were present in the soil of parks, schools and kindergarten recreational areas; the fact may pose a health risk to humans, especially to children, and this type of biological pollution should be regarded as a potential public health concern.

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.

Parasites and fungi as risk factors for human and animal health.

Recent literature data suggests that parasitic and fungal diseases, which pose a threat to both human and animal health, remain a clinical, diagnostic and therapeutic problem. Attention is increasingly paid to the role played by natural microbiota in maintaining homeostasis in humans. A particular emphasis is placed on the possibility of manipulating the human microbiota (permanent, transient, pathogenic) and macrobiota (e.g., Trichuris suis) to support the treatment of selected diseases such as Crohns disease, obesity, diabetes and cancer. Emphasis is placed on important medical species whose infections not only impair health but can also be life threatening, such as Plasmodium falciparum, Echinococcus multilocularis and Baylisascaris procyonis, which expand into areas which have so far been uninhabited. This article also presents the epidemiology, diagnosis and treatment of opportunistic parasitoses imported from the tropics, which spread across large groups of people through human-to-human transmission (Enterobius vermicularis, Sarcoptes scabiei). It also discusses the problem of environmentally-conditioned parasitoses, particularly their etiological factors associated with food contaminated with invasive forms (Trichinella sp., Toxoplasma gondii). The analysis also concerns the presence of developmental forms of geohelminths (Toxocara sp.) and ectoparasites (ticks), which are vectors of serious human diseases (Lyme borreliosis, anaplasmosis, babesiosis), in the environment. Mycological topics contains rare cases of mycoses environmentally conditioned (CNS aspergillosis) and transmissions of these pathogens in a population of hospitalized individuals, as well as seeking new methods used to treat mycoses.

Cytogenetic study of Ascaris trypsin inhibitor in cultured human lymphocytes with metabolic activation

The trypsin inhibitor (ATI) isolated from gastrointestinal nematode Ascaris suum was tested in vitro for induction of chromosome aberrations and sister chromatid exchanges (SCE). Genotoxicity assessment of purified ATI was carried out on metaphase plates received from peripheral blood lymphocyte macroculture (48 h test of structural chromosome aberrations and 72 h test of SCE) with exogenous metabolic activation. ATI was tested in dose of 25, 50 and 100 μg per ml of culture. Kinetics of cell divisions were determined by the replication index (RI). The mitotic index (MI) was expressed as a number of metaphases per 1000 nuclei analysed. Analysis of chromosome aberrations showed that higher doses of ATI (50 and 100 μg/ml) significantly increased the frequency of chromosome aberrations (mainly of chromatid gaps and breaks) compared to the negative control. All concentrations of ATI caused a statistically significant reduction in the MI and RI. In comparison with the negative control, a significant increase in the SCE frequency was observed in all applied doses of ATI. Thus, in the presence of S9 activation, the Ascaris trypsin inhibitor showed potential clastogenic activity and inhibition of the dynamics of lymphocyte divisions.

Prenatal toxicity of Ascaris pepsin inhibitor in mice

The developmental toxicity of pepsin inhibitor isolated from Ascaris suum, a gastrointestinal nematode parasite, was evaluated. An embryo-fetal development study was conducted in BALB/c mice. Groups of 21 mated females were treated by intraperitoneal injection (0.3 ml/30 g body weight) with 0.9% NaCl solution vehicle or isolated Ascaris pepsin inhibitor (API) at dose levels of 50, 100, 150 or 200mg/kg body weight/day on gestation days (GD) 6-15. Maternal food consumption, body weight, and clinical signs were monitored throughout gestation. Cesarean sections were performed on GD 18 and gravid uterine weight, implantation sites, early and late resorptions, live and dead fetuses were collected. Live fetuses were weighed and examined for external, visceral and skeletal variations and malformations. Maternal body weight gain, gravid uterine weight, food consumption were significantly decreased after injection of higher doses of API (100-200mg/kg/day). All doses of API exhibited an embryotoxic effect (high rate of intrauterine resorption). The percentage of postimplantation loss in the groups with administered API was higher (over 4-11 times) than that in control group. Fetotoxicity was observed in all treatment groups in a dose-related manner and it was evidenced by increased dead fetuses, decreased fetal weight, increased visceral variations and reduced skeletal ossification. Fetal hydronephrosis and internal hydrocephalus were noted at 150, and 200mg/kg/day. In summary, the maternal toxicity no-observed-adverse-effect level (NOAEL) was 50mg/kg/day and the low-observed-adverse-effect level (LOAEL) was 100mg/kg/day under the conditions of this study. However, the developmental toxicity LOAEL was 50mg/kg/day based on decreased fetal body weight and prenatal mortality.

Clinical cases of parasitoses and fungal infections important from medical point of view

Most important infectious diseases which pose a risk to human health and life are associated with parasites transmitted by a variety of arthropod vectors, or from animal to man. Some of these (malaria, toxoplasmosis, leishmaniosis, dirofilariosis, alveococcosis, cystic echinococcosis) still represent a serious public health problem in many regions in the world. This review describes the epidemiological and clinical aspects of important parasitoses and fungal infections from a medical point of view. It should be emphasized that the development of invasive disease depends on both host (susceptibility/resistance) and parasite factors (pathogenicity/virulence); an immunocompromised state can favour opportunistic parasitic infections: toxoplasmosis, cryptosporidiosis, giardiosis, cyclosporidiosis, blastocystosis and strongyloidosis. This article highlights the role of free-living amoebae in the pathogenesis and transmission of human diseases, the high pathogenicity of Echinococcus multilocularis, and the growing importance of ticks as a reservoir and vector for numerous dangerous pathogens (e.g., Borrelia burgdorferi, Anaplasma phagocytophilum, Babesia microti). It also discusses the diagnostic problems of toxoplasmosis including cross-reactions in serological tests and reviews the search for new drugs and vaccines against toxoplasmosis. Attention is increasingly paid to the role played by the human microbiome in maintaining homeostasis and in the development of fungal infections. This review also presents the most common human superficial fungal infections and the role of Candida albicans infection in the pathogenesis of irritable bowel syndrome.