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INTRODUCTION: Leishmaniasis is a vector-borne infection endemic in 98 countries. Its cutaneous form (CL) causes skin lesions on exposed parts of the body and may lead to scarring. The numbers of imported cases of CL are increasing in Europe but the incidence of CL importation in Poland is unknown. MATERIAL AND METHODS: A list of all patients diagnosed with CL at the Department in the years 2005-2017 was obtained. The study presents their data including age, sex, areas visited, purpose of travel, time from the onset of symptoms to correct diagnosis, appearance of lesions, results of impression smears and PCR, and superinfection, if detected, as well as treatment methods and their outcomes. RESULTS: Altogether, 14 cases of cutaneous leishmaniasis were identified. The study demonstrates an increase in the number of cases of imported CL at our center over the last 5 years. The time to correct diagnosis was long in spite of the fact that the lesions had usually an appearance typical of CL. CONCLUSIONS: Intensified education of physicians and travelers, as well as improvement in the access to travel medicine services and to the diagnosis and treatment methods appropriate for CL, are needed in our country. In our opinion, surveillance of leishmaniasis should be introduced in Poland.

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Echinococcus multilocularis is a tapeworm that may cause alveolar echinococcosis (AE), one of the most dangerous parasitic zoonoses. As in the case of some foodborne diseases, unwashed fruits and vegetables contaminated with eggs of E. multilocularis may serve as an important transmission route for this parasite. The aim of this study was to investigate the presence of E. multilocularis DNA in fruit, vegetables, and mushrooms in rural areas of the Pomerania province, Poland (non-endemic territory). In total, 104 environmental fruit, vegetable, and mushroom samples collected in forests, plantations, and kitchen gardens were analysed using nested PCR based on the mitochondrial 12S rRNA gene. E. multilocularis DNA was detected in 6.7 % of the samples tested, which indicated that the environment of the Pomerania province is contaminated with this parasite, creating a potential risk for humans. Therefore, fresh fruit, vegetables, and mushrooms should be washed before consumption. Additionally, the results showed that the level of contamination is significantly lower than in the highly endemic Warmia-Masuria province. The differences in the occurrence of E. multilocularis in the environment of these neighbouring provinces appears to be connected with the general epidemiological situation of these two regions, but further study is required for an exact explanation.

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Echinococcus multilocularis is a tapeworm that may cause alveolar echinococcosis (AE), one of the most dangerous parasitic zoonoses. As in the case of other foodborne diseases, unwashed fruits and vegetables, contaminated with dispersed forms of E. multilocularis, may serve as an important transmission route for this parasite. In this article, we reply to the incorrect interpretation of results of our study concerning the detection of E. multilocularis DNA in fresh fruit, vegetable and mushroom samples collected from the highly endemic areas of the Warmia-Masuria Province, Poland, to dispel any doubts. The accusations formulated by the commentators concerning our paper are unfounded; moreover, these commentators demand information which was beyond the purview of our study. Making generalisations and drawing far-reaching conclusions from our work is also unjustified. The majority of positive samples were found in only a few hyperendemic communities; this information corresponds with the highest number of both infected foxes and AE cases in humans recorded in this area. Our findings indicate that E. multilocularis is present in the environment and may create a potential risk for the inhabitants. These people should simply be informed to wash fruits and vegetables before eating. No additional far-reaching conclusions should be drawn from our data. We believe these commentators needlessly misinterpreted our results and disseminated misleading information. Nevertheless, we would like to encourage any readers simply to contact us if any aspects of our study are unclear.

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The erythrocyte binding ligand 140 (EBA-140) is a member of the Plasmodium falciparum erythrocyte binding antigens (EBA) family, which are considered as prospective candidates for malaria vaccine development. EBA proteins were identified as important targets for naturally acquired inhibitory antibodies. Natural antibody response against EBA-140 ligand was found in individuals living in malaria-endemic areas. The EBA-140 ligand is a paralogue of the well-characterized P. falciparum EBA-175 protein. They both share homology of domain structure, including the binding region (Region II), which consists of two homologous F1 and F2 domains and is responsible for ligand-erythrocyte receptor interaction during merozoite invasion. It was shown that the erythrocyte receptor for EBA-140 ligand is glycophorin C-a minor human erythrocyte sialoglycoprotein. In studies on the immunogenicity of P. falciparum EBA ligands, the recombinant proteins are of great importance. In this report, we have demonstrated that the recombinant baculovirus-obtained EBA-140 Region II is immunogenic and antigenic. It can raise specific antibodies in rabbits, and it is recognized by natural antibodies present in sera of patients with malaria, and thus, it may be considered for inclusion in multicomponent blood-stage vaccines.

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Malaria is, along with tuberculosis and HIV/AIDS, one of the three most dangerous infectious diseases in the world. In the absence of native cases since 1963, malaria has remained in Poland an exclusively imported disease, mainly occurring in people travelling to tropical and subtropical areas for professional reasons. The aim of this study was the epidemiological and clinical analysis of 82 patients admitted to the University Center for Maritime and Tropical Medicine (UCMTM), Gdynia, Poland, with a diagnosis of malaria between 2002 and 2014. The "typical" patient with malaria was male, middle-aged, returned from Africa within the preceding 4 weeks, had not used appropriate chemoprophylaxis, and had not applied nonpharmacological methods of prophylaxis, except for window insect screens. P. falciparum was the most frequent species. The most common symptoms included fever, shivers and intensive sweating, thrombocytopenia, elevated creatinine, LDH, D-dimers and CRP, hepatomegaly, and splenomegaly. Within the analyzed group, severe malaria according to WHO standards was diagnosed in 20.7% of patients. Our report presents analysis of the largest series of patients treated for imported malaria in Poland.

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The aim of this study was to estimate the presence of Echinococcus multilocularis DNA in fruits, vegetables, and mushrooms in rural areas of Varmia-Masuria Province, Poland, which is the region with the highest number of human alveolar echinococcosis (AE) cases in this country. Recovery tests showed that E. multilocularis DNA is detectable in samples contaminated with at least 100 eggs of this tapeworm. In total, 103 environmental fruit, vegetable, and mushroom samples collected in forests, plantations, and kitchen gardens were analyzed using nested PCR assay based on the mitochondrial 12S ribosomal RNA (rRNA) gene. The parasite DNA was detected in 23.3% of the samples. Sequencing confirmed that the obtained PCR products represented E. multilocularis. This study is the first environmental survey of the presence of E. multilocularis DNA in fruits, vegetables, and mushrooms intended for consumption. The results clearly demonstrate that it may be a direct source of human infections and shows the need to educate the public about the threat, especially people living in at-risk areas.

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BACKGROUND: Malaria is one of the three most dangerous infectious diseases in the world. According to official statistics, there are a few dozen cases in Poland annually while the number of Poles treated abroad or self-treating remains unknown. Poland has been declared to be malaria-free since 1963 and nowadays all cases are imported. The aim of the study is to determine the minimal number of malaria cases in Poles at home and abroad in the last decade. MATERIALS AND METHODS: The medical records of 4,710 patients tested for malaria in the Department of Tropical Parasitology in the years 2003-2012 were analysed. Two spreadsheets were created, which only included people with a history of malaria diagnosed in the reference centre where indirect immunofluorescent-antibody assay (IFA) for Plasmodium falciparum antigen proved positive. The minimum number of Poles who have had malaria at home and abroad was calculated on the basis of positive IFA results; the rate of all treated malaria patients in Poland in relation to those treated in the reference centre and the actual number of Poles with malaria diagnosed at home was calculated. RESULTS: A group of 376 people with positive serologic tests results in indirect immunofluorescent antibody assay with titre ≥ 1:20 were received, including 227 patients with positive serologic results with titre ≥ 1:80. The rate of the overall number of malaria cases in Poland compared to the number of malaria cases in the University Centre for Maritime and Tropical Medicine Hospital was determined as 3.47:1. It was demonstrated that every year at least 174 to 211 Poles staying abroad may suffer from malaria. CONCLUSIONS: This is the first attempt to estimate the minimal number of Poles infected and treated for malaria in Poland and abroad. The estimated number is 8-10 times greater than the number of registered cases in Poland.

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BACKGROUND: Alveolar echinococcosis (AE) caused by Echinococcus multilocularis infections is a dangerous old disease in the Northern Hemisphere. The aim of the paper was to collect and analyze data on human AE in Poland in the last two decades. METHODOLOGY/PRINCIPAL FINDINGS: The sources of data were both the cases officially registered and detected by an active field and laboratory surveillance. The cases were verified by clinical, epidemiological, and laboratory criteria. Altogether 121 human cases of AE were detected. Among these 83 (68,6%) cases were classified as confirmed, 16 as probable and 22 as possible. During the two decades a continuous increase in detection rate was noticed. The cases were 6-82 years old at the time of diagnosis (mean - 47.7 years). Sex ratio M/F was 0.86/1.0. The AE was fatal in 23 (19%) patients (mean age at death - 54.1 years). Family agglomeration of AE was found in 4 foci, involving 9 patients. Seventy six of the cases were diagnosed in an advanced stage of disease. In all cases the liver was the primary location of AE. In 30 (24.8%) patients a spread to other organs was observed. Ninety four of the patients were treated with albendazole. In 73 (60%) patients a surgical operation was performed, including 15 liver transplantations. CONCLUSIONS/SIGNIFICANCE: The studies confirmed that AE is an emerging disease in Poland, which is the fourth country in Europe with over 120 cases detected. The results also indicate the need of a wider national programme for implementation of screening in the highest AE risk areas (north-eastern Poland) with an effort to increase the public awareness of the possibility of contracting E. multilocularis, and above all, training of the primary care physicians in the recognition of the risk of AE to allow for an early detection of this dangerous disease.

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Dengue is a viral disease caused by an RNA virus of the genus Flavivirus, family Flaviviridae, occurring as four serotypes (DEN-1, -2, -3, -4). It is transmitted to humans by the Aedes mosquitoes, mainly A. aegypti. The occurrence of dengue is strictly related with their preferred breeding areas. Dengue endemic regions are inhabited by some 2.5 billion people. 50-100 million cases of dengue fever and up to 1 million cases of dengue haemorrhagic fever are noted worldwide in more than 100 countries every year. The aim of the reported examinations was to diagnose dengue virus infections in returning travellers. In the years 2006-2009 serological tests were performed in 753 persons. In the diagnostics we used ELISA to find IgM and/or IgG class of antibodies against dengue virus, rapid immunochromatographic (cassette) test, NS1 viral antigen detection by ELISA, and virus RNA detection by RT-PCR method. IgM or IgG class antibodies, and both classes simultaneously, were detected in 19.8% of the examined cases. The greatest number of infections came from India and the Far East, next from South and Central America, and the smallest number from Africa. Sixteen patients with diagnosed dengue, including three cases of dengue haemorrhagic fever, were hospitalized.

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Leishmania parasites are the etiological agents of leishmaniosis, with severe course and often fatal prognosis, and the global number of cases has increased in recent decades. The gold standards for the diagnosis of leishmaniosis are microscopic examinations and culture in vitro of the different clinical specimens. The sensitivity of these methods is insufficient. Recent development in specific and sensitive molecular methods (PCR) allows for detection as well as identification of the parasite species (subspecies). The aim of the study was to estimate the usefulness of molecular methods (PCR) for detection of Leishmania species and consequently for the implementation of such methods in routine diagnostics of leishmaniosis in Polish patients returning from endemic areas of the disease. In our investigations we used 54 known Leishmania positive DNA templates (from culture and clinical specimens) received from the CDC (Atlanta, GA, USA). Moreover, 25 samples of bone marrow, blood or other tissues obtained from 18 Polish individuals suspected of leishmaniosis were also examined. In PCR we used two pairs of primers specific to the conserved region of Leishmania kinetoplast DNA (kDNA) minicircle (13A/13B and F/R). Using these primers we obtained amplicons in all DNA templates from the CDC and in three Polish patients suspected for Leishmania infection. In one sample from among these cases we also obtained positive results with DNA isolated from a blood specimen which was previously negative in microscopic examinations.

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Toxoplasma gondii infection is one of the most common human zoonosis. Humans acquire toxoplasmosis by consuming raw or undercooked meat, from environments and from via vertical transmission to the foetus through the placenta during pregnancy. In the present study, a total of 784 persons (forestry workers), from north-eastern region of Poland (pomorskie and warminsko-mazurskie provinces), were tested by using commercially available ELISA test (DiaSorin) to determine the frequency of Toxoplasma gondii infection. The study was based on detection of IgG and IgM specific antibodies in collected serum samples. The T. gondii-specific IgG were found in 490 samples (62.5%), while the IgM antibodies were noted only in 18 persons (3.7%). High prevalence of seropositive results in forestry workers may suggest an occupational exposure of this group to T. gondii.

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Nematodes from the superfamily Ascaridoidea (families Anisakidae and Raphidascarididae) are worldwide distributed parasites. Their live cycles include many species of water invertebrates and teleostean fish as intermediate hosts, and fish, sea mammals or fish-eating birds being definitive hosts. Humans can be infected with some of these parasites after consumption of raw or wrongly processed fish. The parasitological investigations of fish (herring, cod and flatfish) from southern Baltic (ICES 24-26) provided in the years 80 and 90 showed their infection with larvae of several anisakid species: Anisakis simplex s. str., Contracaecum osculatum C and Hysterothylacium auctum. Sporadically Pseudoterranova decipiens and Raphidascaris acus were also found. Larvae of Anisakis simplex were noted mainly in herrings, C. osculatum primarily in cods and H. auctum in flounders. Additionally, preserved herrings (marinated, smoked) were also investigated and sporadically live larvae of A. simplex were found. The main etiological agent of human anisakidosis worldwide is A. simplex. Although the live cycle of this nematode cannot be completed in the Baltic Sea--this nematode is brought to the Baltic by infected herring migrating from the North Sea for spawning in coastal waters of the Southern Baltic--the prevalence and intensity of infection with larvae of this nematode species were the highest in fish investigated by us. The results obtained suggest the possibility of the human infection with A. simplex larvae in Poland.

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Filth flies associated with a cattle barn and a municipal landfill were tested positive by combined immunofluorescent antibody and fluorescent in situ hybridization for Cryptosporidium parvum and Giardia lamblia on their exoskeletons and in their guts. More pathogens were carried by flies from the cattle barn than from the landfill; 81% of C. parvum and 84% of G. lamblia pathogens were presumptively viable.

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Chloro-, bromo- and methyl- analogues of 1H-benzimidazole and 1H-benzotriazole and their N-alkyl derivatives have been synthesized and tested in vitro against the protozoa Acanthamoeba castellanii. The results indicate that 5,6-dimethyl-1H-benzotriazole (11) and 5,6-dibromo-1H-benzotriazole (14) have higher efficacy than the antiprotozoal agent chlorohexidine.

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Long term field studies and laboratory experiments demonstrated that synanthropic filth flies can mechanically transmit infectious oocysts of Cryptosporidium parvum, an anthropozoonotic protozoan parasite which significantly contributes to the mortality of immunocompromised or immunosuppressed people. C. parvum oocysts are acquired from unhygienic sources, and can pass trough fly gastrointestinal track without alteration of their infectivity and can be subsequently deposited on visited surfaces. Transmission of the oocysts by adult flies occurs via: (1) mechanical dislodgement from the exoskeleton; (2) fecal deposition; and (3) regurgitation, i.e., vomits. Filth flies can cause human or animal cryptosporidiosis via deposition of infectious oocysts on the visited foodstuf, and the biology and ecology of synanthropic filth flies indicate that their potential for mechanical transmission of C. parvum is high.

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In this survey the use of OptiMAL test for rapid diagnosis of malaria was evaluated. It was proved that this test allowed to diagnose the Plasmodium sp. antigen in 72% of examined blood specimens, 82% for P. falciparum infection and 69% for P. vivax, whereas P. ovale was not detected at all. The test sensitivity depended on the parasitemia level. It showed a sensitivity of 100% for parasitemia density exceeded 1%, 95.4% with the parasitemia ranging from 0.1-0.99%. For lower parasite density, the test's sensitivity was of 32 and 60%. The OptiMAL test showed a 99.1% specificity thus it revealed to be significantly high.

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The flagellate Trichomonas hominis is often identified in human diarrheic stools. Because of the fecal-oral transmission rout, infection with T. hominis is more frequently reported in children than in adults. Although infections with the trichomonads in some world regions are as often as with Giardia, no prevention and optimal treatment are defined. The purpose of our in vitro studies was to examine susceptibility of T. hominis to a newly synthesized imidazole derivative--1-metylo-2-(2,4-dinitrobenzylotio)-1H-imidazol--P27; in order to make a comparison between this new and previously known compounds, chlorhexidine and metronidazole were used. The protozoans derived from the diarrheic stool of adult patient were grown on the liquid Pahm medium and subcultured twice a week. Assays of trichomonad cultures were incubated with one of the following substances: chlorhexidine, metronidazole or P27. For each compound, two different concentrations were applied. After 24h exposition, an antitrichomonal effect of the above substances was assessed, namely a quantity of the surviving trichomonads was microscopically determined and compared with that observed in control cultures. The protozoans showed different susceptibility, depending on kind and concentration of tested compound. In comparison to the control cultures, the reduction in number of surviving protozoans appeared in the assays with metronidazole, chlorhexidine and P27 (to 63%, 52% and 8.7%, respectively). The highest in vitro susceptibility of T. hominis was revealed to a new imidazole derivative P27. The obtained results allow us to suppose that the further studies in this field will be very helpful for explaining the mechanisms of the antitrichomonal activity of the tested compounds as well as for assessment of the influence of various abiotic factors.

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