Acquired resistance or tolerance? - in search of mechanisms underlying changes in the resistance profile of Candida albicans and Candida parapsilosis as a result of exposure to methotrexate.

The increasing prevalence of fungal strains showing acquired resistance and multidrug resistance is an increasing therapeutic problem, especially in patients with a severely weakened immune system and undergoing chemotherapy. What is also extremely disturbing is the similarity of the resistance mechanisms of fungal cells and other eukaryotic cells, including human cells, which may contribute to the development of cross-resistance in fungi in response to substances used in e.g. anticancer treatment. An example of such a drug is methotrexate, which is pumped out of eukaryotic cells by ABC transmembrane transporters - in fungi, used to remove azoles from fungal cells. For this reason, the aim of the study was to analyze the expression levels of genes: ERG11, MDR1 and CDR1, potentially responsible for the occurrence of cross-resistance in Candida albicans and Candida parapsilosis as a result of fungal exposure to methotrexate (MTX). In vitro exposure of C. albicans and C. parapsilosis strains to methotrexate showed a high increase in resistance to fluconazole and a partial increase in resistance to voriconazole. Analysis of the expression of resistance genes showed varied responses of the tested strains depending on the species. In the case of C. albicans, an increase in the expression of the MDR1 gene was observed, and a decrease in ERG11 and CDR1. However, for C. parapsilosis there was an increase in the expression of the CDR1 gene and a decrease in ERG11 and MDR1. We noted the relationship between the level of resistance to voriconazole and the level of ERG11 gene expression in C. albicans. This indicates that this type of relationship is different for each species. Our research confirms that the mechanisms by which fungi acquire resistance and develop cross-resistance are highly complex and most likely involve several pathways simultaneously. The emergence of multidrug resistance may be related to the possibility of developing tolerance to antimycotics by fungi.

Cell pleomorphism and changes in the enzymatic profile of selected Candida albicans strains in interaction with Escherichia coli - pilot study.

Interactions between C. albicans and the microbiota play an important role in maintaining the balance between commensal and pathogenic organisms. Although the exact role of bacteria in reducing the pathogenicity of yeast remains poorly understood, a few examples have been documented so far: probiotics administration effectively reduces the formation of biofilm and bacterial metabolites inhibit the formation of hyphae. The aim of the study was to analyze C. albicans virulence levels based on the changes in the morphological structure and enzymatic profile in experimental cultures mixed with Escherichia coli. Viable cell abundance, cell pleomorphism and enzymatic profile were analyzed in single and mixed cultures (C. albicans + E. coli). The microscope analysis showed a large decrease in the number of viable C. albicans cells in mixed cultures with E. coli from 485.3±132.1 immediately after the establishment of the culture to 238.1±71.2 after an hour of incubation and 24.4±5.4 after 24 h. The length of C. albicans cells differed significantly between the single-species cultures and the mixed cultures for 24 h. Our present findings indicate a significant reduction in the secretion of several enzymes by fungi following contact with E. coli, including acid phosphatase, N-acetyl-ß-glucosaminidase, naphthol-AS-BI-phosphohydrolase and leucine arylamidase. The interactions between fungi and bacteria appear to be extremely complex. On the one hand, during C. albicans with E. coli co-incubation, the bacteria stimulated the elongation of yeast cells, leading to the formation of a filamentous form; however, the number of yeast cells and their enzymatic activity decreased significantly. Therefore, it can be concluded that while E. coli stimulates some pathogenic properties, e.g. cell elongation, it also inhibits other virulence features, e.g. enzymatic activity of C. albicans.

Oral form of auto-brewery syndrome.

Although previous cases of ethyl alcohol production by microorganisms present in the intestines, referred to as auto-brewery syndrome (ABS), have been reported, a recent case in our practice was characterized by the production of alcohol in the oral cavity. Our research indicates that legally significant levels of ethyl alcohol can be detected in exhaled air in cases where there has been no alcohol consumption but where the subject has oral candidiasis. In such cases, following the consumption of foods containing carbohydrates, a fermentation process occurs in the mouth, the first stage of which is glycolysis, proceeding according to the Embden-Meyerhof-Parnas pathway, which is typical in eukaryotes. The main organic substrate in this case is glucose, which is formed in the oral cavity from disaccharides (maltose, sucrose) by the activity of α-amylase. Some mutated fungal strains of the genus Candida acquire the ability to break down sucrose and produce glucoamylase. Glucose is converted into glyceraldehyde 3-phosphate and then into pyruvate. The next stage of fermentation is the decarboxylation of pyruvate into acetaldehyde, a reaction catalyzed by pyruvate decarboxylase. The final stage is the reduction of acetaldehyde to ethanol by alcohol dehydrogenase. Such endogenous production of alcohol can be confused with its consumption, which can cause not only legal, but also social and medical problems.

Evaluation of medical students' knowledge of psychoactive substances in the context of their future role in addiction prevention and therapy.

This study assesses the knowledge of medical students on the health effects of the use of psychoactive substances, in the context of their future role in prevention and treatment of addictions. The study was conducted using a questionnaire containing questions about classification, symptoms and effects of psychoactive substances, and the existing prevention programs. The study involved 430 students of medicine and allied faculties. Only 20.8% of medicine students and 12.5% of students of other faculties could correctly classify different psychoactive substances. Correct symptoms of drug misuse were mentioned by 20.4% of medicine students and 19.2% of students of other faculties. The overall knowledge of medical students was no greater than the knowledge of students of other allied faculties. Medical students showed insufficient knowledge about psychoactive substances and their effects on the human body, thus indicating the need to introduce into their study programme a more teaching in this area.

Flexibility in Coping with Stress Questionnaire: structure and psychometric properties.

BACKGROUND: Flexibility in coping relies on applying a coping strategy that is appropriate to the circumstances. Individuals who are flexible in coping exhibit sensitivity to subtle signals sent by the environment. Therefore, they are able to adjust their behaviour accordingly and function in a more adaptive manner. PARTICIPANTS AND PROCEDURE: The study included N = 1535 participants. The following methods were used: the Flexibility in Coping with Stress Questionnaire (FCSQ-14) and the Cognitive Flexibility Inventory (CFI). RESULTS: The Flexibility in Coping with Stress Questionnaire measures flexibility as a feature of the process of coping with which an individual applies coping strategies. The Flexibility in Coping with Stress Questionnaire is an accurate and reliable method of measuring coping flexibility and three of its subscales: the Repertoire of coping strategies, the Changeability of their use, and Reflexivity. CONCLUSIONS: The Flexibility in Coping with Stress Questionnaire examines three subscales of flexibility, namely, Repertoire of coping strategies, Changeability, and Reflexivity. This measurement is accurate and reliable. The questionnaire is applied to the study of persons exposed to severe or chronic stress at work, when we want to assess the individual's ability to change their functioning in a stressful situation. It can be used to examine healthy and somatically ill people, in the area of research and clinical diagnosis.

The occurrence of potentially pathogenic filamentous fungi in recreational surface water as a public health risk.

Microfungi occurring in surface water may represent an important health risk. Recreational water reservoirs are a potential reservoir of pathogenic fungi. The aim of the study was to assess the diversity of mycobiota in selected artificial bathing reservoirs with regard to its biosafety for the human population. The studies were conducted during the summer of 2016 in three research seasons (June (I), July and August (II), and September (III)), taking into account the various periods of recreational activities. Filamentous fungi were isolated from water samples collected at five different ponds utilized for recreation. From 162 water samples, 149 fungal taxa of filamentous fungi were identified: 140 were classified to species level and only nine to genus level. Aspergillus fumigatus was the dominant species. The highest species richness (S) was noted in June, with 93 fungal taxa (Menhinick's index from 2.65 to 4.49). Additionally, in season I, the highest diversity of fungal species was revealed (Simpson's diversity index from 0.83 to 0.99). The average number of CFU/1 mL sample ranged between 0.4 and 4.6 depending on the time of sampling and ponds. Of all the isolated species, 128 were clinically relevant (11 from RG-2 and 117 from RG-1), highlighting the need to introduce seasonal mycological monitoring of such reservoirs.

Evaluation of the relationship between the IL-17A gene expression level and regulatory miRNA-9 in relation to tumor progression in patients with non-small cell lung cancer: a pilot study.

A pro-inflammatory cytokine, IL-17A, is associated with increased risk of developing numerous cancers, including non-small cell lung cancer (NSCLC). IL-17A is a target gene for miR-9. This encouraged us to analyze these two genes in terms of their usefulness as prognostic markers in NSCLC. The expression levels of IL-17A gene and miR-9 was assessed in 26 NSCLC tissue samples and 26 unchanged lung tissue adjacent to lung tumors (control tissue), using qPCR. In both tissue groups, a decreased expression of IL-17A was observed in 100% of samples. Increased expression of miRNA-9 was observed in 92% of tumor samples, and in 100% of control samples. Neither statistical differences in the level of expression IL-17A depending on the patient's age, gender, smoking status, nor histopathology of the cancer was found. Regarding the presence of nodule metastasis ('N' value in TNM classification), significantly lower expression level of IL-17A was observed in cN2 as compared with cN1 group. Additionally, statistically lower IL-17A expression was found in III versus II tumor stage (cAJCC classification). Significant negative correlation between both studied genes was revealed in SCC subgroup. This leads to the conclusion that miRNA-9 can regulate the expression of IL-17A as an IL-17A mRNA antagonistic mediator. Inhibition of proinflammatory action of IL-17A in correlation with tumor progression can be related to various activity of Th17 cells on cancer development according to its immunogenicity, and also may suggest suppressive role of IL-17A in tumor progression. However, because of low number of analyzed samples, further studies on the functional role of IL-17A in development and/or progression NSCLC seem warranted.

An assessment of the relationship between the expression of CCR7/CCL19 axis and selected regulatory miRNAs in non-small cell lung cancer.

CC chemokine receptor type 7 (CCR7) and its ligands has been implicated in the occurrence and progression of NSCLC. Previous studies have revealed that the diagnostic value of CCR7/CCL19 axis in lung tumorigenesis remains controversial. The present study evaluates the relationship between the mRNA expression of CCR7/CCL19 axis and selected regulatory miRNAs in NSCLC patients. It analyzes the expression level of CCR7 mRNA and its ligand in tumor tissue in relation to expression level of two miRNAs: miR let-7a and miR-335, as transcriptional regulators of study genes. Twenty-seven patients (n = 27) were enrolled. The expression of the studied genes and miRNAs was evaluated by qPCR. Tumour tissue fragments, adjacent macroscopically-unchanged lung tissue (control) and patient serum were used as biological material for study. Elevated expression of CCR7 and CCL19 mRNA was observed in patients with metastasis to lymph nodes. We noticed upregulated miR-335 expression and downregulated miR let-7a expression in patient serum with regard to AJCC tumor staging. Higher miR-335 expression and lower miR let-7a expression level was observed in patients with metastasis to lymph node. The presence of changes observed in the expression level of miR-335 and miR let-7a in the serum of NSCLC patients in relation to lymph node metastases and tumor stage may serve as a non-invasive molecular biomarker of tumor progression; however, this observation requires further investigation.

Neuroinfections caused by fungi.

BACKGROUND: Fungal infections of the central nervous system (FIs-CNS) have become significantly more common over the past 2 decades. Invasion of the CNS largely depends on the immune status of the host and the virulence of the fungal strain. Infections with fungi cause a significant morbidity in immunocompromised hosts, and the involvement of the CNS may lead to fatal consequences. METHODS: One hundred and thirty-five articles on fungal neuroinfection in PubMed, Google Scholar, and Cochrane databases were selected for review using the following search words: "fungi and CNS mycoses", CNS fungal infections", "fungal brain infections", " fungal cerebritis", fungal meningitis", "diagnostics of fungal infections", and "treatment of CNS fungal infections". All were published in English with the majority in the period 2000-2018. This review focuses on the current knowledge of the epidemiology, clinical presentations, diagnosis, and treatment of selected FIs-CNS. RESULTS: The FIs-CNS can have various clinical presentations, mainly meningitis, encephalitis, hydrocephalus, cerebral abscesses, and stroke syndromes. The etiologic factors of neuroinfections are yeasts (Cryptococcus neoformans, Candida spp., Trichosporon spp.), moniliaceous moulds (Aspergillus spp., Fusarium spp.), Mucoromycetes (Mucor spp., Rhizopus spp.), dimorphic fungi (Blastomyces dermatitidis, Coccidioides spp., Histoplasma capsulatum), and dematiaceous fungi (Cladophialophora bantiana, Exophiala dermatitidis). Their common route of transmission is inhalation or inoculation from trauma or surgery, with subsequent hematogenous or contiguous spread. As the manifestations of FIs-CNS are often non-specific, their diagnosis is very difficult. A fast identification of the etiological factor of neuroinfection and the application of appropriate therapy are crucial in preventing an often fatal outcome. The choice of effective drug depends on its extent of CNS penetration and spectrum of activity. Pharmaceutical formulations of amphotericin B (AmB) (among others, deoxycholate-AmBd and liposomal L-AmB) have relatively limited distribution in the cerebrospinal fluid (CSF); however, their detectable therapeutic concentrations in the CNS makes them recommended drugs for the treatment of cryptococcal meningoencephalitis (AmBd with flucytosine) and CNS candidiasis (L-AmB) and mucormycosis (L-AmB). Voriconazole, a moderately lipophilic molecule with good CNS penetration, is recommended in the first-line therapy of CNS aspergillosis. Other triazoles, such as posaconazole and itraconazole, with negligible concentrations in the CSF are not considered effective drugs for therapy of CNS fungal neuroinfections. In contrast, clinical data have shown that a novel triazole, isavuconazole, achieved considerable efficacy for the treatment of some fungal neuroinfections. Echinocandins with relatively low or undetectable concentrations in the CSF do not play meaningful role in the treatment of FIs-CNS. CONCLUSION: Although the number of fungal species causing CNS mycosis is increasing, only some possess well-defined treatment standards (e.g., cryptococcal meningitis and CNS aspergillosis). The early diagnosis of fungal infection, accompanied by identification of the etiological factor, is needed to allow the selection of effective therapy in patients with FIs-CNS and limit their high mortality.

Neuroinvasions caused by parasites

Parasitic diseases of the central nervous system are associated with high mortality and morbidity. Many human parasites, such as Toxoplasma gondii, Entamoeba histolytica, Trypanosoma cruzi, Taenia solium, Echinococcus spp., Toxocara canis, T. cati, Angiostrongylus cantonensis, Trichinella spp., during invasion might involve the CNS. Some parasitic infections of the brain are lethal if left untreated (e.g., cerebral malaria ­ Plasmodium falciparum, primary amoebic meningoencephalitis (PAM) ­ Naegleria fowleri, baylisascariosis ­ Baylisascaris procyonis, African sleeping sickness ­ African trypanosomes). These diseases have diverse vectors or intermediate hosts, modes of transmission and endemic regions or geographic distributions. The neurological, cognitive, and mental health problems caused by above parasites are noted mostly in low-income countries; however, sporadic cases also occur in non-endemic areas because of an increase in international travel and immunosuppression caused by therapy or HIV infection. The presence of parasites in the CNS may cause a variety of nerve symptoms, depending on the location and extent of the injury; the most common subjective symptoms include headache, dizziness, and root pain while objective symptoms are epileptic seizures, increased intracranial pressure, sensory disturbances, meningeal syndrome, cerebellar ataxia, and core syndromes. Many early symptoms of CNS invasion are often nonspecific therefore a diagnosis can be difficult. This article presents the epidemiology, pathophysiology and clinical manifestations of selected parasitic neuroinfections.

Sandpits as a reservoir of potentially pathogenic fungi for children.

INTRODUCTION AND OBJECTIVE: Fungi belonging to various physiological and morphological groups present in the environment are potential human pathogens. Some of them are considered as emerging pathogens. Therefore, their presence in children's playgrounds should be regarded as health risk factor. MATERIALS AND METHOD: Sixty-eight samples of sand collected from 17 sandpits of different localities in Lódz, Poland, in autumn 2010 and 2011, and in spring 2011 and 2012 were evaluated. The fungi were isolated with classical mycological methods and identified on the basis of morphological and biochemical features. RESULTS: The prevalence of fungi in spring was 94.1% of sandpits in both layers of sand (depth 0-3 cm and 10-15 cm) and in one kindergarten sandpit, but only in a deeper layer. In autumn, fungi occurred in both layers in all sandpits (100%). The fungal concentration (CFU/g of sand) varied considerably (range 0 - uncountable) in both layers. A total of 352 isolates belonging to 80 species were found. There were 69 yeasts and yeast-like fungi isolates from 12 species (9 species in each season), and 283 filamentous fungi from 68 species: 35 species in spring and 55 in autumn, with 4 keratinolytic species. There were important causes of allergies, among them Cladosporium herbarum and Alternaria alternata, as well as of opportunistic mycoses: Cryptococcus neoformans, Aspergillus fumigatus and new and 'emerging' fungal pathogens e.g., Trichosporon, Rhodotorula, Fusarium and Scedosporium species. CONCLUSIONS: Potentially pathogenic fungi are present in the sand taken from sandpits in Lódz. This fact poses a significant threat to child health and therefore proper maintenance and periodic checking of sandpits are of great importance.

The hydrolytic enzymes produced by fungi strains isolated from the sand and soil of recreational areas

The pathogenicity of fungi depends on, inter alia, the secretion of hydrolytic enzymes. The aim of this study was to determine the enzymatic activity of yeasts and yeast-like fungi isolated from children's recreation areas, and compare the results with literature data of strains obtained from patients with mycoses. The enzymatic activity of 96 strains was assessed using an API ZYM kit (bioMerieux, France) and their biotypes were established. The fungal species were found to produce from 16 to 19 hydrolases: the most active were: leucine arylamidase (e5), acid phosphatase (e10), alkaline phosphatase (e1), naphthol-AS-BI-phosphohydrolase (e11), esterase ­ C4 (e2), ß-galac - tosidase (e13) and ß-glucosidase (e16). In addition, 13 biotypes characteristic of particular species of fungi were defined. Most strains could be categorized as biotypes C2 ­ 39.5% and A ­ 26%. The examined fungal strains isolated from recreational areas have selected biochemical characteristics i.e. production of hydrolases, which demonstrate their pathogenicity. They produce a number of enzymes which are also present in strains isolated from patients with mycoses, including: leucine arylamidase (e5), acid phosphatase (e10), naphthol-AS-BI-phosphohydrolase (e11) and alkaline phosphatase (e1). The biotypes identified in the course of this study (A, B3, B4, C1, C6 and D3) have been also reported in cases of fungal infection. Therefore, the fungi present in the sand and soil of recreational have pathogenic properties and are possible factors of fungal infection among children.

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.

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 children's 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 Lódz, 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 children's 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.

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 Crohn's 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.

Interactions between potentially pathogenic fungi and natural human microbiota.

The human body is composed of 1014 cells, of which only 10% of them belong to the human host itself:the remaining 90% are microorganisms. Commensal microorganisms are necessary for the proper functioning of the human body and covers an area that could potentially become sites of adhesion of pathogenic microorganisms, it thus represents a form of competition for potential pathogens. The coexistence of fungi and bacteria in cases of systemic infections is a significant diagnostic and therapeutic problem, and the human immune system reacts differently,depending on the pathogen. Numerous publications exist concerning the relationship between microorganisms belonging to different ecological groups, the majority of which concern the interaction between macro-organisms and potential pathogens, or the synergistic relationship between parasitic species. However, there is still too little information concerning the role of natural microbiota in maintaining homeostasis and the relationships between particular species inhabiting the human organism.

Assessment of the applicability of a "toolbox" designed for microbially assisted phytoremediation: the case study at Ingurtosu mining site (Italy).

The paper describes the fieldwork at the Italian test site of the abandoned mine of sphalerite and galena in Ingurtosu (Sardinia), with the aim to assess the applicability of a "toolbox" to establish the optimized techniques for remediation of soils contaminated by mining activities. A preliminary characterization-including (hydro)geochemistry, heavy metal concentration and their mobility in soil, bioprospecting for microbiology and botany-provided a data set for the development of a toolbox to deliver a microbially assisted phytoremediation process. Euphorbia pithyusa was selected as an endemic pioneer plant to be associated with a bacterial consortium, established with ten selected native strains, including metal-tolerant bacteria and producers of plant growth factors. The toolbox was firstly assessed in a greenhouse pot experiment. A positive effect of bacterial inoculum on E. pithyusa germination and total plant survival was observed. E. pithyusa showed to be a well-performing metallophyte species, and only inoculated soil retained a microbial activity with a high functional diversity, expanding metabolic affinity also towards root exudates. These results supported the decision to proceed with a field trial, investigating different treatments used singly or in combination: bioaugmentation with bacterial consortia, mycorrhizal fungi and a commercial mineral amendment. Microbial activity in soil, plant physiological parameters and heavy metal content in plants and in soil were monitored. Five months after the beginning, an early assessment of the toolbox under field conditions was carried out. Despite the cold season (October-March), results suggested the following: (1) the field setup as well as the experimental design proved to be effective; (2) plant survival was satisfactory; (3) soil quality was increased and bioaugmentation improved microbial activity, expanding the metabolic competences towards plant interaction (root exudates); and (4) multivariate analysis supported the data provided that the proposed toolbox can be established and the field trial can be carried forward.

Parasites and fungi as a threat for prenatal and postnatal human development.

Recent literature data reveals the most common etiological agents of congenital parasitoses to be Toxoplasma gondii, Trypanosoma cruzi, Leishmania donovani and Plasmodium falciparum. An analysis of clinical data indicates that parasitic congenital infections are often asymptomatic, whereas symptomatic newborns usually display nonspecific symptoms, which greatly hinders correct diagnosis. The long-term consequences of prenatal infections are serious clinical problems. This article presents the possible routes of vertical transmissions (mother-to-child) of pathogens including prenatal, perinatal, as well as postnatal routes. It highlights the role of factors involved in protozoa transmission and development of congenital parasitic diseases, such as parasite genotypes, the relationship between the timing of maternal infection and the probability of passage of the parasite through the placental barrier, and the immunological features of pregnant women. Acquired and congenital babesioses in human and experimental animals are presented. It emphasises that the mechanisms by which parasites infect the placenta and cross from mother to fetus are still poorly understood. It also describes the cellular mechanisms of infection by T. gondii, such as tachyzoites crossing biological barriers, the expression of Toll-Like Receptors (TLR) family on trophoblasts and syncytiotrophoblasts as an immune response to intrauterine infection and cases of congenital and acquired toxoplasmosis, as well as the long-term consequences of congenital invasion with T. gondii, episodes of reactivation of latent toxoplasmosis and T. gondii reinvasions. Mycological topics include a rare case of in utero fungal infection of offspring by a mother with vaginal candidosis, and the fungal contamination of ward facilities and medical equipment as potential sources of exogenous infections of newborn children.

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.