Annals of Parasitology 2012, 58(3), 111–123 Copyright© 2012 Polish Parasitological Society

Review articles

Current problems concerning Parasitology and Mycology with regard to diseases of the skin and its appendages 1

Joanna Błaszkowska, Anna Wójcik

Chair of Biology and Medical Parasitology, Medical University of Lodz, 1 Hallera Square, 90-647 Lodz, Poland

Corresponding author: Joanna Błaszkowska; E-mail: [email protected]

ABSTRACT. Current issues concerning Parasitology and Mycology with regard to diseases of the skin and its appendages are presented. Aspects of diagnostics, clinical picture and therapy of skin and nail mycoses, as well as difficulties in the diagnosis and treatment of both native parasitoses (toxoplasmosis) and imported human tropical parasitoses (malaria, filariosis) have been emphasised. The clinical importance of environmental mould fungi in nosocomial infections and fungal , as well as selected properties of fungi isolated from patients with head and neck neoplasms treated by radiotherapy are discussed. Other mycological topics include the characteristics of newly- synthesized thiosemicarbazides and thiadiazoles as potential drugs against toxoplasmosis and their biological activity against Toxoplasma gondii tachyzoites, selected molecular mechanisms of resistance to azoles, strains and a new tool (barcoding DNA) for describing the biodiversity of potential allergenic . The importance of environmental factors in pathogenesis of mycoses and parasitoses is noted. The characteristics of pathogenic fungi isolated from natural ponds in Bialystok and potentially pathogenic -like fungi isolated from children’s recreation areas in Lodz are presented. The ongoing problem of anthropozoonoses is considered, as are the roles of stray cats and dogs in contaminating soil with the developing forms of intestinal parasites. The characteristics of the human microbiome, including population composition, activity and their importance in normal human physiology, are presented, as are the major goals of the Human Microbiome Project initiated by National Institutes of Health (NIH).

Key words: medical parasitology, mycology, parasitoses, mycoses During the last decades, a permanent increase in The spectrum of isolated from the number of fungal skin infections has been skin lesions has changed in the last 70 years in most observed worldwide. The high prevalence of European countries. and superficial mycoses shows that approximately , ranked highest before 20–25% of the world’s population has skin lesions, the Second World War in Germany, have been making these one of the most frequent forms of replaced by rubrum , which nowadays infection. Pathogens responsible for those diseases accounts for 80–90% of the strains, followed by are primarily geophilic, anthropophilic and T. mentagrophytes . This trend, also observed in the zoophilic dermatophytes from the genera Central and Northern Europe, is directly connected Trichophyton , Microsporum and Epidermophyton . with the increased incidence of tinea pedis. In There appears to be considerable inter and intra- contrast, in Southern Europe and in Arabic continental variability in the global incidence of countries, zoophilic dermatophytes, such as these fungal infections. Dermatophytes are or , keratinophilic and keratinolytic fungi, characterized are the most frequently isolated. In Europe, by a high affinity to keratin-containing tissues. In especially in Mediterranean countries, the incidence everyday clinical practice they are responsible for of M. canis infection has strongly increased during superficial mycoses of skin (, tinea the recent years and this is now the barbae, , , tinea manuum or most prevalent in in children [3]. tinea pedis), nails (, tinea unguium) Infections caused by dermatophytes have and (tinea capitis) [1,2]. become a very serious problem, not only a clinical

1 The review was supported by Medical University of Lodz, No 503/1-013-01/503-01 112 J. B³aszkowska, A. Wójcik one, but also with epidemiological and therapeutic smaller toenails was induced by T. mentagrophytes aspects. Clinical manifestations of mycoses vary and T. rubrum , whereas of the large toenail depending on the site of infestation and the type of was often associated with Candida species and also strain and therefore, accurate identification of the moulds of genera Fusarium , Alternaria , strain is crucial in order to facilitate rapid treatment , Scopulariopsis [4] . Interestingly and to prevent spread of the disease. Skin mycoses enough, S. brevicaulis , a common saprophytic often present a typical clinical picture, however, in found in soil, on vegetables and other some cases they may mimic other disorders, such as organic waste, having a wide geographic pityriasis rosea Gibert, granuloma annulare, distribution, may be an etiological factor of skin sarcoidosis, urticaria, erythema annulare lesions in various locations and also generalized centrifugum, lupus erythematosus (especially mycoses. Several case reports have described subacute cutaneous form – SCLE), erythema S. bre vicaulis infections of deep tissue or skin in chronicum migrans, erythema multiforme, immunocompromised hosts and there have been psoriasis, eczema and contact dermatitis. Also, the sporadic cases of ocular infection. A case of slowly previous application of some drugs (such as progressive granulomatous face skin infection in an steroids, antihistamines, antibiotics or zinc otherwise healthy person has also been found [5]. ointment) may modify the presentation of Onychomycosis, infection of the nail plates, is dermatomycoses in some patients. Therefore, it is of caused by dermatophytes, nondermatophytes great importance to collect scrapings for moulds or yeast species. In the majority of patients, microscopy and culture before starting treatment onychomycosis is viewed as more than a mere with oral or topical antifungals (J. Narbutt; Medical cosmetic problem. In spite of improved personal University of Lodz). hygiene and living environment, nail mycoses The commonest risk factors for fungal skin continue to spread and persist. It has been estimated infections are: antibiotic and corticosteroid use, a that nail mycoses may concern nearly 30% of adults depressed immune system due to chemotherapy and in Europe, whereas their prevalence in Australia and HIV infection, the coexistence of diabetes, as well the United States accounts for 10% [6]. as environmental factors like increased moisture, Numerous studies have revealed that nearly 10% lifestyle, contact with soil, form of recreation and of the cases of onychomycosis are caused by taking part in different sports and, in some nondermatophyte molds (NDM): Scopulariopsis individuals, a genetic predisposition towards brevicaulis , Aspergillus spp ., Acremonium spp., mycoses. Religious ceremonies and traditions, like Fusarium sp ., Onychocola canadiensis , Nattrasia ritual ablutions, are also of the great importance. mangiferae , Alternaria spp ., Scytalidium dimidia - The relative occurrence of the etiologic agents and tum , S. hyalinum , or Clado- fungal skin infections varies from country to spo rium carrionii . Unlike dermatophytes, those country and from one climatic region to another. In fungi are not keratolytic and can only invade a nail tropical countries, a warm and humid climate, plate previously destroyed due to trauma, crowded living and poor sanitary conditions all dermatophyte activity or another nail disease. For promote the spread of mycoses. It is worth noting this reason, the incidence of onychomycosis due to that mycoses of the skin covering the trunk and face NDM is estimated from 1.45 to 17.6% [7,8]. have been more frequently observed in those Keratinophilic molds may be either countries, whereas in a moderate climate, fungi contaminants of biological material in the mostly affect the feet and hands, particularly the laboratory or pathogens causing nail lesions, which nails. In Lodz (Poland), a higher prevalence of skin may create serious problems for the correct and nail lesions was noticed in middle-aged and diagnosis of NDM onychomycosis. Moreover, elderly patients as compared to young subjects, with diagnostic criteria are inconsistent between significantly greater occurrence on the hands of mycological laboratories. Based on several studies, women than men (J. Kwaśniewska, E. Szefer, six major diagnostic criteria were identified: A. Jaskółowska; Medical University of Lodz). identification of the NDM in the nail by Mycosis of the large toenail was most frequently microscopy, the presence of mycelium elements diagnosed, while infection of the remaining (using a potassium hydroxide preparation – KOH toenails, foot skin and spaces between toes occurs with Parker ink or Calcofluor White; alternatively more rarely. In the majority of cases, mycosis of the KOH may be replaced by Na 2S or NaOH), isolation Current problems in Parasitology 113 in culture, repeated isolation in culture in different management of Candida related CNS infections time intervals, quantitative evaluation of inoculum, [10]. failure to isolate a dermatophyte in culture and A unique case of fungal meningitis with many histology (periodic acid Schiff). Most studies years’ course, probably stemming from a recommend using at least three of those criteria to communication injury, is also described. The patient rule out contamination, always including the KOH was repeatedly hospitalized between 2006 and 2012 preparation for direct microscopy and isolation of in the Department of Infectious Diseases in the organism in culture (P. Krzyściak; Jagiellonian response to a fever above 38 °C, headache, nausea University-Medical College). Of note, mixed nail and vomiting. The patient was discharged several invasions are possible and dermatophytes may not times from the hospital on his own request before be isolated till the second or the third culture. If the finishing the diagnostic and therapeutic procedures. same mould is still present in subsequent cultures, it In this case, C. glabrata was found to be responsible should be regarded as the etiological factor of for the meningitis; it was also isolated from the onychomycosis [9]. nasopharyngeal cavity with a negative culture from Fungi from the genus Candida , although urine and feces specimens. The most reliable uncommon, may be etiological factors for explanation is that fungal meningitis is connected meningitis. Mostly the disease affects immuno- with a previous injury and untreated sinusitis, which suppressed patients, those treated with broad- eventually leads to quantitative changes in the spectrum antibiotics or those receiving parenteral T lymphocyte CD4 count and the spread of nutrition, or it can be the result of disseminated C. glabrata from nasopharynx towards the central disease. In addition, two specific patient groups, nervous system. It is recommended that every premature neonates and neurosurgical patients, are lymphocytic meningitis of unknown origin should at increased risk. Several studies based on autopsies be carefully screened for fungal etiology, especially have proved that the true prevalence of fungal in case of depressed cellular immunity (A. meningitis can be underestimated in everyday Michowicz, M. Jabłkowski; Medical University of clinical practice. Fungal infection of the central Lodz). nervous system manifests as meningitis, micro or During the last decade, invasive infections macro-abscesses, vascular and medullar injury. caused by Candida albicans have been revealed as C. albicans accounts for 70–100% of all fungal an increasing problem especially in immuno- isolates, but an increase in the occurrence of other compromised patients. Of many antifungal agents, species, such as C. glabrata , C. tropicalis , C. para - only very few substances are established as agents psilopsis and C. lusitaniae , has been observed. in the treatment of systemic fungal infections. Infections caused by non albicans species, are Azoles, especially , have been used as frequently resistant to antifungals and are associated first-line antimycotics for the prophylaxis and with higher mortality. In contrast to meningitis treatment of fungal infections. As the number of caused by C. albicans , C. tropicalis meningitis has patients treated with fluconazole has increased over been increasingly reported in adults. Most cases of the last 15 years, azole resistance has emerged as a C. tropicalis are postoperative complications of major problem. The azoles bind to and inhibit the head and neck surgery, including mastoid activity of lanosterol 14 α-demethylase (Erg11p), a exploration, craniotomy, and ventricular-peritoneal key enzyme in the fungal ergosterol biosynthesis shunt. The diagnosis of meningitis is established by pathway. The development of clinically resistant repeated positive CSF cultures. CSF parameters are Candida strains can occur by several mechanisms, variable, with a mild lymphocytic or polymor- including infection with intrinsically resistant phonuclear pleocytosis and an increased protein organisms, selection of resistant organisms level. Fungal elements are generally not seen. secondary to antifungal drug pressure, and Hence, CSF abnormalities are indistinguishable development of resistance in a previously from cryptococcal, tuberculous, and some bacterial susceptible organism. Several mechanisms of meningitides. It is of great concern that despite resistance to azoles have been described in C. appropriate treatment, this entity is still associated albicans [11]. These include increased expression with 10–30% mortality, and the percentage of of drug efflux pump genes such as MDR1, CDR1 neurological sequels ranges from 18 to 29%, which and CDR2, amino acid substitutions in the target highlights the importance of timely recognition and enzyme Erg11p due to mutations in the ERG11 114 J. B³aszkowska, A. Wójcik gene, as well as overexpression of ERG11 [12]. whe re as in fec tions due to Asper gil lus spp. ac co unt Overexpression of efflux pump genes such as for 1.3%. Ho we ver, the epi de mio lo gy has chan ged MDR1 and MDR2 leads to a decreased intracellular dra ma ti cal ly in re cent years; in va si ve fun gal in fec- azole concentration. In addition, mutations in the tions cau sed by mo ulds, pre do mi nan tly Asper gil lus ERG11 gene encoding the target enzyme lanosterol spe cies, ha ve in cre ased sub stan tial ly and new ly 14 α-demethylase prevent the binding of azoles to emer ging fun gal pa tho gens such as Zy go my ce tes the active site. (e.g. Rhi zo pus , Mu cor , Ab si dia ), hy ali ne mo ulds In a presented study by a team from the Medical (e.g. Fu sa rium ) and other op por tu ni stic spe cies (e.g. University of Silesia (K. Gołąbek, J. Strzelczyk, A. Sce do spo rium ) are in cre asin gly be ing re por ted Owczarek, A. Wiczkowski), the levels of expression [13,14]. Ac cor ding to stu dies con duc ted on pa tients of genes encoding ERG11 and efflux transporters with he ma to lo gic ma li gnan cies du ring a 15-year pe- (MDR1 and CDR1, CDR2) were monitored in riod (1989–2003) a high pre va len ce of in va si ve fun- susceptible and resistant isolates of C. albicans . A gal in fec tion (31%) was found at autopsy, and range of samples comprising 120 strains of although 77% of patient deaths were related to C. albicans was isolated from hospitalized patients infection, only one-third of these infections were in the Independent Public Hospital in Nowy Targ. diagnosed by the accepted criteria before the The antifungal susceptibility of C. albicans isolates patients died. Several cancer centers have reported samples to fluconazole, and an increase in the incidence of infections caused by was evaluated using ATB fungus2 difficult-to-treat opportunistic molds such as (bioMérieux). Total RNA was extracted from Zygomycetes , Fusarium , and Scedosporium species. C. albicans cells by use of the RNeasy® Mini Kit Moreover, Trichoderma longibrachiatum was (Qiagen, Germany) and was characterised with the recently described as an emerging pathogen in RNA LabChip Kit by 2100 Bioanalyzer (Agilent immunocompromised patients, specifically in a Technologies, USA). A quantitative analysis of the renal transplant recipient [15]. An invasive mould level of expression of the examined genes was infection is associated with a high rate of mortality. performed by RT-Q-PCR. In this study, increased Also, mould fungi from Microascales (Scopu la - expression of the MDR1 and CDR2 genes was riopsis brevicaulis , and discovered in resistant Candida strains. Significant Scedosporium prolificans ) are the cause of differences were observed regarding ERG11 nosocomial infections. is an important expression in the tested azole-resistant and azole- cause of morbidity and mortality, particularly in sensitive strains of Candida . These results suggest allogeneic transplant recipients and neutropenic that overexpression of MDR1, CDR2 and ERG11 patients with hematologic malignancies. More than genes can be an important mechanism of azole 90% of fusariosis cases have been reported in resistance in C. albicans clinical isolates. neutropenic patients with hematologic mali - Hospital-acquired infections can be caused by gnancies. with Scedosporium sp. was viruses, bacteria, fungi or parasites. Among the noted in 11% of solid organ transplant recipients various fungi which may cause nosocomial [16]. A potential reservoir of airborne moulds is the infections, the most common are mould species, indoor air of hospital buildings. Disease can be which mostly infect immunosuppressed individuals transmitted by the inhalation of spores in air, (A. Gniadek, A.B. Macura; Jagiellonian University- consumption of contaminated water and by direct Medical College). It was estimated that the inoculation on disrupted skin or mucosa. incidence of nosocomial infections varies between 5 Sporangiospores released by moulds range from 2.5 and 10% with mortality calculated approximately to to 40 m in diameter, are easily aerosolized, and are be 11%. The wider use of more aggressive methods readily dispersed throughout the environment. of treatments, such as chemotherapy, radiotherapy, Respiratory tract colonization due to inhalation of or gan trans plan ta tion, long term ad mi ni stra tion of the mould depends on the size of the spores. Large cor ti co ste ro ids and im mu no sup pres si ve the ra py spores of Alternaria (23–34 μm) settle in the pro mo te no so co mial in fec tion s; fun gal in fec tions nosopharynx, but very small spores ( Rhizopus 4–6 re pre sent ap pro xi ma te ly 9–10% of all no s o co mial μm, Mucor 4–8 μm, A. fumigatus 2.5–3 µm, in fec tions. It is com mon know led ge that Can di da Fusarium 2.4–3.5 μm) can reach the lungs and enter spe cies are the most com mon fun gal pa tho gens cau- the alveoli. Hospital building works, failing to clean sing no so co mial in fec tions (85% of all my co sis) airflow systems and high-efficiency particulate air Current problems in Parasitology 115

(HEPA) filters increase the risk of invasive mould patients treated by radiotherapy emphasises the infections. necessity to perform mycological examinations in Radiotherapy is one method of treatment used in this group of patients. cancer therapy which can injure the oral, pharyngeal Endocrinopathies and coexisting mycoses or laryngeal mucosa, and predispose the tissue to the concern not only humans: animals can also be development of fungal infections [17]. As seen from affected. In recent years, an increasing frequency of the literature data, the prevalence of fungal infection fungal infections in cats and dogs has been observed has increased in patients treated by chemo- and [20,21]. The likely predisposition to infection was radiotherapy, and it may affect as many as 3/4 of the thought to be local or systemic immune patients involved [18]. A determination of the compromise. It was revealed that Candida albicans , frequency of fungal infection in particular phases of C. guillermondi , C. humicola and M. pachydermatis radiotherapy, and selected pathogenicity parameters were isolated more frequently from dogs and cats of the strains has great significance in treatment and with diabetes, hypothyroidism and Cushing disease prognosis. In a study conducted by Moqbil and compared with healthy animals. All isolated Kurnatowski (Medical University of Lodz), fourty- samples were obtained from the oral cavity and three patients with oral, pharyngeal or laryngeal rectum (W. Dardzińska, B. Dworecka-Kaszak; carcinoma were subjected to mycological Warsaw University of Life Sciences). examinations at four points during a course of Although much biological research depends radiotherapy: prior to the course, in week 3, on the upon species diagnoses, there is a considerable last day of the course and 3 weeks after treatment. crisis in taxonomic research at the moment. It is The fungal strains were isolated and identified from believed that the sole prospect for a sustainable the oral cavity of these patients. A number of means of identification lies in the construction of phenotypic characteristics, such as the activity of systems that employ DNA sequences as taxon fungal hydrolytic enzymes, adherence properties of „barcodes”. The standard methods used in strains to epithelial cells and sensitivity to mycological diagnostics, based on phenotypic antifungal drugs, were determined. The presence of features, are characterized by 70–85% sensitivity fungi in the mouth and throat was found in over 2/3, and specificity, implying that almost 15–30% of 4/5, about 3/5 and all of the patients in the first, the species could not be correctly identified. This second, the third and the final examination, situation may especially concern Candida non- respectively. The most frequent species isolated albicans species involved in the increasing number from the patients was Candida albicans (40–60%) of various, sometimes generalized, infections which and multidrug-resistant C. glabrata . The isolated are resistant to standard antifungals. Therefore, strains were the most sensitive to nystatine and molecular methods including barcoding may help to miconazole, and in the least sensitive to precisely identify the fungus. Nevertheless, it ketoconazole and fluconazole. Regarding the should be remembered that molecular methods, particular biotypes for different species of Candida , such as PCR with specific and universal primers, the most significant diversity (classification for analysis of the length of PCR product, restriction A–D biotypes) was demonstrated for C. albicans fragment length analysis, hybridization with and C. glabrata [19]. The isolated strains are specific probes and DNA sequence analysis also characterized by the highest activity of leucine have their limitations and always have to be arylamidase, acid phosphatase and naphthol- AS- preceded by basic procedures used in all BI-phosphohydrolase. Considering the enzymes mycological laboratories [22]. produced, most of the strains can be included to The ide al DNA bar co de cha rac te ri zes low in tra- biotypes D 3 (22.5%), C 6 (17.5%) and A (15.0%) spe cies (~2%) with si mul ta ne ously high in ter spe- and less frequently to C 3 (7.5%), B 1 (7.5%) and B 2 cies va ria bi li ty, flan king by con se rva ti ve do ma ins (1.3%). The adherence ability was found to range and short length (up to 600 ba se pa irs). Ana ly sis of from 0 to 57 fungal cells to one cell of the mouth the ITS re gion (in ter nal trans cri bed spa cer) has pro- epithelium. Fungal cells demonstrated increasing ved to be ef fec ti ve in the iden ti fi ca tion of spe cies adherence to the epithelial cells of the mouth in the wi thin a num ber of ge ne ra: Pe ni ci llium , Asper gil lus , three subsequent batches (prior to, week 3, last day Fu sa rium , Rhi zo pus , Uloc la dium , Sco pu la riop sis of radiotherapy). The high prevalence of fungi and My ce lia ste ril la . The re was a fa ilu re in the pre- revealed in this study in the mouth and throat of ci se se qu en ce ana ly sis of clo se ly re la ted Pe ni cil lium 116 J. B³aszkowska, A. Wójcik spe cies ( Pe ni ci llium fre ii/P. po lo ni cum , P. ochro - ly stok). In autumn 2011and spring 2012, wa ter sam- chlo ron/P. pu lvil lo rum , P. bia lo wie zien se/P. bre vi- ples we re col lec ted from three ponds wi thin the com pac tum) , but in such ca ses, the β-tu bu lin re gion town of Bia ły stok: Fo sa Pond, Doj li dy Pond and was used ef fec ti ve ly. The ITS re gion al so de mon- Ko mo sa La ke. Wa ter sam ples for exa mi na tion we re stra ted so me li mi ta tions du ring the dia gno stics of ob ta ined 0.20 m un der the sur fa ce of the wa ter using fun gi from Cla do spo rium and Al ter na ria ge ne ra. Rut t ner‘s ap pa ra tus (2.0 dcm 3 ca pa ci ty). A ba it me- Ad di tio nal ly, trans la tion elon ga tion fac tor 1- α was thod using se eds as ba it was ap plied to iso la te fun- spe ci fic for Cla do spo rium and Fu sa rium, but not for gi. The sam ples we re sto red for ap pro xi ma te ly one Al ter na ria (K. Ple wa, E. Lonc; Uni ver si ty of Wroc - month at a tem pe ra tu re of 18 °C. The ba it was suc- law). ces si ve ly ob se rved un der an opti cal mi cro sco pe and yeast-like fungi are important (100 and 400x ma gni fi ca tion) eve ry 3–5 days, star- components of many ecosystems and possess ting from day 3 of the cul tu re. The fun gi we re iden - economic, agricultural and medical importance ti fied ba sed on the fol lo wing mor pho lo gi cal fe atu- through their considerable impact on fundamental res: the sha pe and si ze of the thal lum, the sha pe of soil processes, such as decomposition, aggregation, the spo ran gium and spo res, the struc tu re of the nutrient release and storage, as well as their oogo nium, ga me tan gium and oospo ra. The se mi cro- interaction with vegetation and soil animals. The sco pi cal ly clas si fied my ce lia we re re mo ved from species composition of different localities is highly the se eds and trans fer red to ste ri li zed Pe tri di shes heterogeneous and the quantities fungi range from con ta ining Sa bo uraud’s me dium or Po ta to De xtro se hundreds to millions of cells per gram of soil. Up to Agar (PDA). This stu dy re ve aled the oc cur ren ce of 130 species have been reported from soils 30 spe cies from 5 or ders: Chy tri dio my ce tes (5), Zy- worldwide [23,24]. go my ce tes (1), Oomy co ta (19) and ana mor phic fun- Fungi may become potential human and animal gi (5). The most com mon spe cies we re Apha no my - pathogens, especially for individuals with a ces la evis , Ca te no ph lyc tis va ria bi lis and Sa pro le- depressed immune system. The presence of fungi in gnia fe rax . Three new spe cies we re di sco ve red: the environment, especially in soil, may be a factor Achlya dif fu sa , Py thium gra ci le , Sa pro le gnia li to ra - favouring their spread into human ontocenoses. An lis. The fol lo wing po ten tial pa tho ge nic fun gi we re evaluation of soil samples ta ken from child re cre- de ter mi ned: La ge ni dium gi gan teum , Ca te no ph lyc tis ation si tes in Lodz in au tumn 2010 and spring 2011, va ria bi lis , Rho do to ru la glu ti nis , Al ter na ria al ter na ta , re ve aled a va rie ty of spe cies: Can di da col li cu lo sa , Cla do spo rum her ba rum , Asper gil lus can di dus , Pe ni - C. gu il lier mon dii , C. hu mi co la , C. in con spi cua , ci llium chry so ge num. The iso la tion of an th ro po pa - C. lam bi ca , C. lu si ta niae , C. pel li cu lo sa , C. tro pi ca- tho ge nic fun gi from wa ter sam ples sug ge sts that na - lis , Cryp to coc cus al bi dus , C. lau ren tii , C. neo for - tu ral aqu atic re se rvo irs are an im por tant so ur ce of mans , C. ter reus , Klo ec ke ra ja po ni ca , Geo tri chum fungal infections. These species can be potentially can di dum , G. pe ni cil la tum , Rho do to ru la ru bra , applicable as an indicator of water contamination, R. glu ti nis , Sac cha ro my ces ce re vi siae , Spo ro bo lo- and can be useful in sanitary-hygienic assessment. my ces sal mo ni co lor and Tri cho spo ron cu ta neum. The WHO defines zoonoses as diseases and The most com mon so ur ce of Cryp to coc cus iso la tes infections that are naturally transmitted between we re the so il of sand pits, scho ol pit ches and park vertebrate animals and humans. Protozoan and play gro unds. Mo re over, Cryp to coc cus neo for mans , helminth zoonoses are of great importance in public an etio lo gi cal fac tor of cryp to coc cal me nin gi tis, was health. In urban areas, the main source of many al so pre sent in the sand pits of 3 kin der gar tens. The parasitoses is soil contaminated with feline or Can di da fun gi we re iso la ted in so il sam ples from all canine faeces. The most frequent intestinal parasites unfen ced park play gro unds and unfen ced scho ol pit- of dogs are helminths e.g. Toxocara canis , ches on ly in au tumn. The num ber of co lo ny for ming Toxascaris leonina , Ancylostoma caninum , units (cfu/1g of so il) of all iso la tes was gre ater in au - Uncinaria stenocephala , Trichuris vulpis and tumn than spring- ti me (A. Wój cik, J. Błasz kow ska, Dipylidium caninum. Nematodes, like Toxocara P. Kur na tow ski, K. Szwa be; Me di cal Uni ver si ty of cati , Toxascaris leonina and Ancylostoma Lodz UM). tubaeforme are common parasites of the alimentary The cha rac te ri stics of pa tho ge nic fun gi iso la ted tracts of cats. Most of them can infect humans from three na tu ral ponds in the Bia ły stok area was [25,26], but T. canis and T. cati have the greatest pre sen ted (B. Ki zie wicz, Me di cal Uni ver si ty of Bia- epidemiological importance [27]. The prevalence of Current problems in Parasitology 117 intestinal parasite infections in dogs and cats from Surveillance Network, a global network of the Lodz Shelter for Animals was assessed (K. medicine/travel clinics, was established in 1995 to Szwabe, J. Błaszkowska; Medical University of detect morbidity trends among travellers. Filariosis Lodz). A total of 137 fecal samples from 87 dogs comprised 0.62% (n = 271) of the 43 722 medical and 50 cats were examined by a sedimentation conditions reported to the GeoSentinel Network method using a Mini Parasep faecal parasite between 1995 and 2004. Immigrants from sub- concentrator. The overall prevalence of intestinal Saharan Africa were most likely to have acquired a parasites in dogs and cats were 35.8% and 58%, filarial infection than other travellers and long-term respectively. Five different parasite species were travel (greater than 1 month) was more likely to be observed in stray dogs, and eight species were associated with acquisition of filariosis than shorter- detected in samples from the cats. In dogs, the term travel [28]. prevalence of parasites found were: Toxocara canis A case was described of a 50-year-old man, who (18.1%) , Uncinaria stenocephala (8.1%), Trichuris was admitted to the Department of Infectious vulpis (5.8%), Capillaria spp. (2.3%) and Isospora Diseases in May 2008 with cellulitis, panniculitis spp. (1.2%). In cats, the following species were and lymphangitis of the left crus with moderate detected: Toxocara cati (32%), Ancylostoma fever. All symptoms had started in February 2008. caninum/Uncinaria stenocephala (2%), Capillaria The patient had been suffering from chronic spp. (2%), Dipylidium caninum (6%), Taenia headache, which had lasted for many years without taeniaeformis (2%), Spirometra erinacei (2%), any reaction towards non-steroidal anti- Isospora spp. (16%) and Giardia felis (2%). inflammatory drugs or carbamazepinum. He had Toxocara was the most frequently found worm travelled twice to India in the 1980s and Africa genus in dogs as well cats. In 20% of canine (Namibia) in 2005. The initial diagnosis was samples mixed infections were found ( T. canis and erysipelas, eosinophilia was not noticed and patient Uncinaria stenocephala ; Trichuris vulpis and received the standard treatment. In June 2008, the Uncinaria stenocephala ; T. canis , Uncinaria patient was readmitted to the hospital due to renal stenocephala and Capillaria sp.). In the examined and liver failure, bronchitis, left thigh elephantiasis cat population, more than one parasite were and gonocele. No fever, leukocytosis or observed in 8% of fecal samples ( T. cati and eosinophilia were noticed. Thick and thin Isospora sp., Giardia felis and Isospora sp.). The smears, haematoxylin stained, revealed the presence results obtained in this study are evidence that the of microfilariae probably connected with greater part of animals adopted by the Lodz Shelter W. bancrofti infection. The second case concerned a were infected with intestinal parasites. Parasite 58-year-old patient with generalized myalgia and eggs, cysts or oocysts present in faeces can be arthralgia, weakness and enlargement of neck transmitted to the soil and are a potential source of lymphatic nodes lasting or less than six months. human infections. This patient had been in Kenya, where he had gone Filariosis is a parasitic disease caused by thread- on safari, for 14 days almost 7 years before. There like nematodes, transmitted by blood-feeding were no changes in general and serological tests and arthropods: mainly black flies and mosquitos. The as in the first case, blood smears revealed parasites. disease can be generally divided into three main These presented cases underline the necessity of types: lymphatic (caused by Wuchereria bancrofti , blood smear evaluation in patients with a suspicion Brugia malayi and Brugia timori , where worms are of parasitic disease and no eosinophilia, even if it found in the lymph nodes, what can lead to has been many years since they might have travelled elephantiasis), subcutaneous (caused by Loa loa , to endemic parts of the word before the present Mansonella streptocera and Onchocerca volvulus , symptoms occurred (Z. Deroń; Bieganski occupying the subcutaneous layer of the skin, Voivodship Specialist Hospital in Lodz). involving in river blindness) and serous cavity Malaria, a mosquito-borne infectious disease of disease (caused by Mansonella perstans and humans and other animals caused by protozoan M. ozzardi discovered within the abdomen with parasites of the genus Plasmodium , is still a major hydrocele). As international travel increases, there global problem, causing about 250 million clinical is a rising exposure to many pathogens not cases a year. It is responsible for nearly 800 000 traditionally encountered in the resource-rich deaths a year, almost all in people living in poverty countries of the world. The GeoSentinel with limited access to healthcare, but it also causes 118 J. B³aszkowska, A. Wójcik avoidable deaths every year from imported malaria which was the only chance to make a correct in non-endemic countries. Such a situation is strictly diagnosis, was not performed. After a consultation related to the increased rate of travel to endemic with the infectious disease specialist the patient was areas and a lack of knowledge about the risk of discharged from the hospital with antiprotozoal malaria , both by tourists and their physicians. This drugs eliminating the liver reservoir, but the is the main reason why, in many European treatment was delayed. After 4 months, the patient countries, there is no pre-travel advice and therefore was readmitted to the hospital due to fever. Despite no diagnosis and proper treatment upon return. The a confirmed plasmodium infection, he was not number of cases of imported falciparum malaria has properly treated and serious complications, increased over the past four years in the United including heart failure and encephalopathy, Kingdom with 5774 reported cases between 2007 occurred. In 2007, the patient experienced another and 2011. Most travelers acquiring malaria are of relapse. Thick and thin blood smears once again African heritage visiting friends and relatives. In confirmed recurrent malaria caused by P. vivax . contrast, the risk of dying from malaria once Treatment with arechin interrupting the attacks, acquired is highest in the elderly, tourists and those followed by primachin, proved to be effective. This presenting in areas in which malaria is seldom seen case indicates the need for blood smear evaluation [29]. when malaria is suspected, followed by treatment of Between January 1999 and September 2003, a the attacks and eradication of the liver reservoir. total of 4 801 patients with travel-related malaria Toxoplasma gondii is a prevalent obligate were reported within the TropNetEurop Network. intracellular parasite which has chronically infected P. vivax was present in almost 13% of cases, either more than a third of the world’s population. More as the sole pathogen or in mixed infections with recently, it has been learned the T. gondii infection other species, thus accounting for the second highest has the ability to modify host behaviour [31]. A number of cases after P. falciparum . The reported correlation has been identified between latent proportion of P. vivax remained steady with 10.9% toxoplasmosis and human behavioral changes such in 1999, 12.6% in 2000, 15.1% in 2001, 12.3% in as slower reactions, lower rule consciousness and 2002 and 12.9% in 2003. Of note, Germany increased activity as well as deficits in learning (24.3%), Spain (15.5%) and the UK (12.0%) capacity and memory. Studies show that these reported most cases, whereas reports from parasites may be a causative or contributory factor Switzerland (1.8%), Poland (1.6%), Finland (1.0%), in such psychiatric disorders as depression, anxiety, Ireland (1.0%) and Portugal (0.3%) were scarce. schizophrenia, Parkinsons disease and epilepsy According to diagnostic information, 557 (90.1%) [32,33]. Behavioral changes in mice infected by of the 618 infections were confirmed, two (0.3%) Toxoplasma gondii were estimated (J. Gatkowska, were probable and eight (1.3%) were suspected. The M. Wieczorek, B. Dzia dek, K. Dzit ko, H. Dłu goń- remaining 51 (8.3%) could not be classified due to ska; Uni ver si ty of Łodz). The expe ri ments we re per- missing information on the underlying diagnostic for med on C57BL/6 in bred mi ce, which are in na te - procedure [30]. ly hi gh ly su scep ti ble to T. gon dii . The in ter me dia te Plasmodium vivax infections are characterized vi ru lent ME 49 stra in of T. gon dii was used to in du- by relapses of malaria arising from persistent liver ce expe ri men tal to xo pla smo sis. Mi ce we re in fec ted stages of the parasite (hypnozoites) which can be by in tra pe ri to ne al ad mi ni stra tion of 20 cy sts per prevented only by anti-malarials. A pertinent case mo use. The be ha vior of non- in fec ted mi ce and mi ce is that of a 72-year-old patient, an internal medicine in fec ted acu te ly (3 we eks p.i.) and chro ni cal ly (6 specialist, who was admitted to the Department of we eks p.i.) with T. gondii was quantified in an open Infectious Diseases in 2004 due to moderate fever field (OF) using a 59×59×30 cm box with its floor (Z. Deroń; Bieganski Voivodship Specialist divided into center and peripheral parts illuminated Hospital in Lodz). He have had a malaria in youth by red light. Each mouse was placed in the center of while working in Georgia. Initial diagnostic the arena facing the wall, then videotaped for 5 min procedures confirmed only caries. The patient was and scored with the EthoVision system. Three given arechin, as if during a malaria attack, with groups of behavioral parameters were measured: 1/ transient improvement. Anti- Plasmodium anti - exploratory activity (rearing and climbing on the bodies were negative, but a basic screening test walls), 2/ spontaneous locomotor activity (entries to including evaluation of thick and thin blood smears, the central and peripheral parts of the OF arena), Current problems in Parasitology 119 and 3/ anxiety-like behavior (self-grooming). T. gondii in L-929 cell line. The DL 50 values of the Infected mice showed significantly diminished majority of tested compounds for intracellular exploratory activity described by climbing and tachyzoites were lower than the DL 50 of rearing, reduced preference for the central, more sulfadiazine (DL 50 > 500 g/ml). These chemicals exposed part of the OF arena and engaged in less were not seen to have any influence on extracellular grooming behavior compared to the uninfected tachyzoites. Of the newly synthetized thiose - controls. The locomotor activity of infected mice micarbazides and thiadiazoles, the highest anti- was most pronounced in the peripheral part of the Toxoplasma effect was demon strated by 4-(4- open field especially during acute parasitosis. This fluorophenyl)-1-(tiofen-2-yl)-carbony lthio semi - study confimed that the presence of T. gondii cysts carba zide (DL 50 =33.17 g/ml). in the brain caused behavioral changes in mice. The The de tec tion of To xo pla sma -spe ci fic an ti bo dies cysts were detected during late acute (3 weeks p.i.) is the pri ma ry me ans of dia gno sing in fec tion with and chronic (6 weeks p.i.) toxoplasmosis both in the To xo pla sma gon dii. Se ve ral te sts to iden ti fy the hippocampus and the amygdala of infected mice. pre sen ce of im mu no glo bu lin IgG ha ve be en in tro- According to some investigators, these structures of du ced to help di scri mi na te be twe en re cen tly acqu- the brain are involved in defense behaviors, such as ired and di stant in fec tion. To xo pla sma an ti bo dy de - conditioned or learned fear, and nonconditioned tec tion te sts are per for med by a lar ge num ber of la- anxiety [34,35]. bo ra to ries with com mer cial ly ava ila ble kits. A com - In fec tion with the pro to zo an To xo pla sma gon dii pa ra ti ve eva lu ation of fo ur To xo pla sma -spe ci fic cau ses se rio us pu blic he alth pro blems and is of gre- IgG an ti bo dy te sts, (three com mer cial and one not), at eco no mic im por tan ce worl dwi de. The stan dard and the ir si gni fi can ce in dif fe rent cli ni cal set tings the ra py of to xo pla smo sis is the com bi na tion of py - was con duc ted by E. Go łąb, G. Kar czew ski and M. ri me tha mi ne plus sul fa dia zi ne or clin da my ci ne. Ho- Ba kal- No wak (NIZP -PZH, LUX MED; LIM Me di - we ver, this tre at ment is often as so cia ted with se ve re cal Cen tre, War saw). Se ven ty se rum sam ples we re si de ef fects in c lu ding al ler gic re ac tions and bo ne exa mi ned to iden ti fy the pre sen ce of im mu no glo bu- mar row sup pres sion. For this re ason, the re is an on- lin IgG by the fol lo wing fo ur te sts: 1/ che mi lu mi ne - go ing se arch to find new drugs to co un ter this form scent mi cro par tic le im mu no as say (CMIA) – To xo of pa ra si to sis. Thio se mi car ba zo ne and thia dia zo le IgG, Ar chi tect Sys tem (Ab bott Dia gno stics Di vi - de ri va ti ves are of con si de ra ble in te rest be cau se of sion), 2/ an en zy me -lin ked flu ore scent as say (EL - the ir, among others, an ti pro to zo al, an ti bac te rial and FA) (VI DAS To xo IgG, bioMérieux), 3/elec tro che- an ti tu mor ac ti vi ties. The an ti -To xo pla sma ac ti vi ty mi lu mi ne scen ce as say Co bas (Ro che Dia gno stics of new ly syn the si zed thio se mi car ba zi des and GmbH), 4/ in di rect im mu no flu ore scent- an ti bo dy as- thiadiazoles have been estimated (K. Dzitko, A. say - IFAT (Na tio nal In sti tu te of Pu blic He alth -Na- Ruszczak, A. Siwek, H. Długońska; University of tio nal In sti tu te of Hy gie ne, Po land). An ti bo dy le vels Lodz, Medical University of Lublin). In preliminary we re eva lu ated by fol lo wing the ma nu fac tu rer’s in- studies, these compounds were tested for in vitro struc tions whe re ap pro pria te. In this stu dy, an in i tial cytotoxicity, using mouse L929 fibroblasts and de tec tion of IgG an ti bo dies to T. gon dii was per for- HeLa cells by 3-(4,5-dimethylthiazol-2-yl)-2,5- med with the CMIA – To xo IgG, Ar chi tect Sys tem diphenyltetrazolium bromide (MTT) assay. kit. Of 70 se rum sam ples, 20 we re ne ga ti ve, 49 sam- Tetrazolium salts are widely used as indicators of ples we re po si ti ve and one was a do ubt ful ly po si ti - metabolic activity for both eukaryotic and ve re sult. All ne ga ti ve se rum sam ples we re con fir- prokaryotic cells. Live cells reduce the tetrazole ring med by other ap plied te sts (cri te ria for de ter mi ning in the MTT salt, forming a colored formazane which of ne ga ti ve re sults: for Ar chi tect To xo IgG as say can be assessed spectrophotometrically. In the and IFA test – IgG le vel < 1.6 IU/ml, for Co bas < present study, the MTT test was used to assess the 1.0 IU/ml, Vi das < 4.0 IU /ml). The se rum sam ple survival of extracellular tachyzoites in other con si de red do ubt ful ly po si ti ve by con sen sus was fo- concentrations of samples. In addition, [3H] uracil und to be in de ter mi na te with the CMIA; this sam ple incorporation was used to evaluate the efficacy of was al so fo und to be do ubt ful by VI DAS, but po si - new compounds against the proliferation of ti ve by Co bas and IFAT. Of the 49 po si ti ve sam ples, T. gondii in vitro in mouse L-929 fibroblasts. These no dif fe ren ces we re fo und be twe en the VI DAS and results showed that the tested thiazolidinone and IFAT test re sults with re gard to me an I.U. of IgG/ml thiadiazole derivates decreased proliferation of of se rum ob ta ined, but the se dif fe ren ces we re ob se- 120 J. B³aszkowska, A. Wójcik rved in sam ples exa mi ned by the Co bas and Ar chi- Leech includes three species: Hirudo medicinalis , tect te sts. The par tial agre ement of the re sults from H. verbana and H. troctina. However, due to the dif fe rent com mer cial as says in di ca tes that the se hirudotherapy complications such as infections by te sts are unre lia ble in dia gno sing T. gon dii in fec tion microorganisms, anaphylaxis, local allergic when se rum im mu no glo bu lin IgG con cen tra tions reactions, anemia and cellulitis can occur. A high are low. It is worth em pha si zing that se rum IgG an- incidence of Aeromonas infection after application ti -To xo pla sma go dii an ti bo dy ti tres (expres sed in of medicinal leeches, despite their external I.U.) are de pen dent on the me thod of me asu re ment decontamination, necessitates an antibiotic used. Hen ce, on ly one test sho uld be used to mo ni- prophylaxis. For this reason, an important clinical tor To xo pla sma -spe ci fic IgG le vels in the pa tient. and biological aspect of medical leeches is the Le eches are ec to pa ra si tes on va rio us ver te bra tes, identity of the microbiota in the digestive tract and in c lu ding hu mans. After the le ech at ta ches with its also on body surface of these animals. In a study suc kers to the host, the sa li va ry glands se cre te a conducted by Litwinowicz and Błaszkowska num ber of com po unds in c lu ding an ane sthe tic, a va- (Medical University of Lodz), a mycological so di la tor, a spre ading fac tor and pro te oly tic in hi bi- assessment was performed of the purity of the outer tors. A le ech can suck and sto re blo od up to ten ti- coating of medical leeches, as well as their jaws, mes its bo dy si ze. They can stay ali ve for abo ut two other parts of their intestine, and also purity of water years with blo od sto red in the ir crop. It has be en in which the leeches were kept. The study was shown that bac te ria, vi ru ses, and pa ra si tes from pre- performed on 20 Hirudo verbana obtained from the vio us blo od so ur ces can su rvi ve wi thin a le ech for BIO-GEN company. Each leech was washed in 5 ml mon ths, and may be trans mit ted to a hu man host. of Sabouraud’s broth to rinse potential fungi out of The in ge sted blo od in the le ech di ge sti ve tract co uld the cover-muscular sack. Cultures were incubated at be re- in jec ted in to the host, along with the va rio us 37 °C for 48–72h. The jaws, crop and intestine of pa tho gens, by re gur gi ta tion du ring the ma ni pu la tion leeches were incubated in Sabouraud’s broth at of le ech re mo val. Le ech bi tes ha ve be en as so cia ted 37 °C for 48–72h. After centrifugation of 50 ml with Ae ro mo nas ve ro nii bio var so bria in fec tions, samples, 1ml of water were taken, inoculated on a com pli ca ting up to 20% of me di cal le ech the ra pies, Petri dish with solid Sabouraud’s medium and then but mi ght al so be in vo lved in the pa tho ge ne sis of incubated for 48–72 h at 37 °C. Mycological va rio us tro pi cal so il -re la ted in fec tions, such as diagnostics were conducted using morphological Chro mo bac te rium vio la ceum sep ti ca emia and chro- and biochemical characteristics of the isolated mo bla sto my co sis [36]. Mo re over, in ter nal hi ru di no- fungi. Fungi were found in all samples from the ses ha ve be en described in the literature. Sites of cover-muscle sack, the jaws and samples of water. leech infestation include the nasal cavities, pharynx, In total, 18 species of fungi from 8 genera were larynx, respiratory tract, , urinary bladder and isolated in the study. No fungi were recorded in the rectum [37]. A rare case of an intraperitoneal intestine. The following species: Candida krusei , hirudinosis in a 2-year-old girl has been described , Rhodotorula rubra, Tricho - [38]; the leech entered her vagina and uterus, sporon asahii and asteroides were perforated the uterus and entered into the peritoneal isolated only from water samples. C. albicans was cavity. The presence and survival of pathogens found in all tested materials. There was detected inside the gut of leeches has been studied [39]. HIV fungi classified as potential pathogens (BSL-2): and hepatitis-B virus, bacteriophages and bacteria C. albicans , C. krusei , C. ciferrii , C. tropicalis , have been found to persist in large numbers for at T. asahii and T. asteroides as well as species with a least 6 months in the gut of experimentally infected lowered potential of pathogenicity (BSL-1), such as leeches. Protozoan parasites such as Toxoplasma C. guilliermondii , C. parapsilosis or saprophytic gondii , Trypanosoma brucei brucei , or Plasmodium fungi: Lipomyces starkeyi, Rhodosporidium sp., berghei were even capable of reproducing inside the Trichosporonoides sp. Isolation of anthropo- gut of the leech, and they survived about 5–6 weeks. pathogenic fungi from the bodies and jaws of Leeches are currently in use in Medicine, medical leeches, and the water they live in, suggests especially in plastic and reconstructive surgery, that they are an important source of fungal wound and flap healing, in venous insufficiencies, infections in patients exposed to hirudotherapy. and in the treatment of many disorders such as Humans live in constant association with the hemorrhoids and varicosity. The European Medical microorganisms present on the surfaces and in the Current problems in Parasitology 121 cavities of the human body, and even within cells. rather than the result of exposure to a rare pathogen. Over 500 species form the complex and dynamic The knowledge of the microbiome enables an communities which constitute the human insight into areas of diet and body chemistry, and microbiome (microbiota). The largest collection of hence, into their associated fundamental metabolic microbes resides in the gut (10–100 trillion pathways (A. Jaworski; University of Lodz). organisms). The gut microbiome genes and Homo In the available literature, there are several sapiens genes together form the human studies concerning treatment using fecal „metagenome”, which can have a considerable bacteriotherapy, a treatment first described in 1958 impact on human physiology, nutrition and and now often referred to as fecal microbiome immunity, and may be a significant factor in the transplantation. During such a procedure, the bowel pathophysiology of many diseases. The intestinal is cleaned with antibiotics and then infused with microbial community is inherited from the mother. donor-processed fecal suspensions daily for 5 to 15 Knowledge of the functional genomics of microbial days. After 24 weeks, the donor flora is relatively communities will allow the characterization of stable in the microbiota of the feces. This method is molecular signals and the mechanisms of the suggested to be effective for new therapies in the dialogue between microorganisms and their host. treatment of colonic or metabolic disease. Other The creation of probiotic and prebiotic preparations studies evaluated the treatment of Clostridium specific for the aged population is one example of difficile diarrhea by fecal transplantation, resulting the medical/nutritional applications whose in therapeutic success after 14 days, which was then development has been spurred on by the growing seen to last for almost 4 years. Microbiome body of evidence concerning the importance of gut transplantation method is still very controversial microbiota homeostasis to the health of the host and not accepted by Food and Drug Administration [40,41]. (FDA) and other institutions responsible for Establishment of the intestinal microbiota has introducing new medical procedures worldwide been shown to be a progressive process. The major [43]. functions attributed to it begin to manifest at the end The occurrence of yeast–like fungi and yeasts in of the second year of life and comprise: nutrients the oral cavity is regarded by some authors as a absorption and food fermentation, stimulation of the physiological state, and by others as a sign of the host immune system and barrier effects against violation of immunological barriers. According to pathogens. Once climax composition is achieved the literature, the prevalence of fungi in the oral near the end of adolescence, this ecosystem displays cavity can vary between 10 and 96% [44]. The most a high stability in healthy adults but recent studies commonly isolated fungi from the genus Candida , indicated that modifications occur in the although in recent years, the frequency of the genus composition in elderly individuals [42]. Saccharomyces has increased. One factor favoring It was revealed that bacteria of the two most the colonization of the mucous membranes by fungi important phylogenotypes of gastrointestinal tract and the development of mycosis is a high- (GI) microbiome, Firmicutes and Bacteroides , do carbohydrate diet. One reason for the greater not have their external reservoir. This observation consumption of monosaccharides is the signifi- proved the theory of vertical transmission of GI cantly greater availability and popularity of various bacteria. Some studies suggest that there must be a types of sweetened beverages. However, infor - pathway of transmitting signals from GI mation about the connection between the microbiome, through L-cells’ receptors localized in consumption of such beverages and the colon secreting peptide hormones, to regions in the development of fungi is scarce. One study central nervous system responsible for investigated the correlation between frequent metabolism’s regulation. consumption of sweetened beverages and increased The Human Microbiome Project (HMP), a risk of fun gal co lo ni za tion in the oral ca vi ty (K. Gó- United States National Institute of Health initiative, ral ska, P. Ja błoń ski, A. Klim czak, P. Ra chu biń ski, has been created in order to examine microbiome D. Tyr na, A. Kwa pi szew ska, A. Ja głow ska, A. Bor- changes and its influence on human health. kow ska, P. Ko za rzew ski; Stu dent’s Scien ti fic Circ- Understanding the nature of health can allow the les be lon ging to Cha ir of Bio lo gy and Me di cal Pa- clinical target of therapy to be more precisely ra sitology of Medical University of Lodz). Students defined. Many illnesses are ecological diseases completed an original questionnaire which 122 J. B³aszkowska, A. Wójcik contained, among other things, information on the Brint nell W., Pi rac ci ni B.M., To sti A. 2012. Sys te ma- type, quality and quantity of beverages consumed. tic re view of non der ma to phy te ony cho my co sis: For mycological studies, oral washings were dia gno sis, cli ni cal ty pes, epi de mio lo gy, and tre at- collected with liquid Sabouraud’s medium, which ment. Jo ur nal of the Ame ri can Aca de my of Der ma to- were then incubated (temp. 37 , 24 h) and direct lo gy 66: 494-502. ° [10] He nao N.A., Va gner B. 2011. In fec tions of the cen- preparations were made. In the case of fungal tral ne rvo us sys tem by Can di da . Jo ur nal of In fec tio us isolation, mycological diagnostics were performed Di se ases and Im mu ni ty 3: 79-84. according to established procedures. The [11] Akins R.A. 2005. An upda te on an ti fun gal tar gets preliminary studies showed that fungi were present and me cha ni sms of re si stan ce in Can di da al bi cans . in the oral cavity of 52% of those examined, and the Me di cal My co lo lo gy 43: 285-318. respondents declaring consumption of sweetened [12] Xu Y., Chen L., Li C. 2008. Su scep ti bi li ty of cli ni- beverages were found to be associated with the cal iso la tes of Can di da spe cies to flu co na zo le and de- increased prevalence of fungi (60–62.5%), tec tion of Can di da al bi cans ER G11 mu ta tions. Jo ur - compared to those who didn’t. The study confirms nal of An ti mi cro bial Che mo the ra py 61: 798-804. that a diet high in sweetened beverages may favour [13] Lass- Florl C. 2009. 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