Imported cutaneous leishmaniasis: a 13-year experience of a Polish tertiary center.

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.

Fresh fruits, vegetables and mushrooms as transmission vehicles for Echinococcus multilocularis in highly endemic areas of Poland: reply to concerns.

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.

The first detection of Echinococcus multilocularis DNA in environmental fruit, vegetable, and mushroom samples using nested PCR.

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.

Dengue antibodies in Polish travellers returning from the tropics. Evaluation of serological tests.

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.

Usefulness of PCR method for detection of Leishmania in Poland.

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.

[Occurrence of Toxoplasma gondii specific antibodies in group of forestry workers from pomorskie and warminsko-mazurskie provinces]. / Wystepowanie specyficznych przeciwcial anty-Toxoplasma gondii w grupie pracowników lesnych z województwa pomorskiego i warminsko-mazurskiego.

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.

Detection of Echinococcus multilocularis DNA in fruit, vegetable, and mushroom samples collected in the non-endemic territory of the Pomerania province and comparison of the results with data from rural areas of the neighbouring highly endemic Warmia-Masuria province, Poland.

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.

Studies on Immunogenicity and Antigenicity of Baculovirus-Expressed Binding Region of Plasmodium falciparum EBA-140 Merozoite Ligand.

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.

Prevalence of anisakin nematodes in fish from Southern Baltic Sea.

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.

An attempt to estimate the minimal number of Poles infected and treated for malaria in Poland and abroad.

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.

Imported Malaria in the Material of the Institute of Maritime and Tropical Medicine: A Review of 82 Patients in the Years 2002-2014.

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.

Molecular confirmation of human alveolar echinococcosis in Poland.

Infections of humans with Echinococcus multilocularis, the causative agent of alveolar echinococcosis (AE), a zoonosis, have been described with increasing frequency in Poland since 1994. In the attempt to verify these reports, we analyzed specimens obtained from a representative group of Polish patients. Liver lesions in patients with AE that was diagnosed on the basis of results of histological and serological tests contained E. multilocularis DNA, as shown by the presence of specific microsatellite sequences and mitochondrial 12S rDNA. The same tests clearly distinguished between AE and cystic echinococcosis, which is caused by Echinococcus granulosus. These data are unequivocal proof that human infections with E. multilocularis occur in Poland.

Evaluation of a novel method based on amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting) for typing strains of vancomycin-resistant Enterococcus faecium.

In the search for an effective DNA-typing technique for use in hospital epidemiology, the performance and convenience of a novel assay based on the fingerprinting of bacterial genomes by amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting) was tested. A large number of vancomycin-resistant Enterococcus faecium (VREM) isolates from haematological ward patients of the Clinical Hospital in Gdansk were examined. We found that ADSRRS fingerprinting analysis is a rapid method that offers good discriminatory power. The method demonstrated also excellent reproducibility. The usefulness of the ADSRRS fingerprinting method for molecular typing was compared with pulsed field gel electrophoresis (PFGE) method, which is currently considered the gold standard for molecular typing of isolates recovered from patients and the environment in the course of investigation and control of nosocomial outbreaks. Clustering of ADSRRS fingerprinting data matched pulsed field gel electrophoresis data. The features of ADSRRS fingerprinting technique is discussed in comparison with conventional methods. Data presented here demonstrate the complexity of the epidemiological situation concerning VREM that may occur in a single medical ward.

[Risk of alveococcosis for humans in Poland]. / Zagrozenie bablowica wielojamowa (alweokokoza) dla ludzi w Polsce.

The paper presents data on occurrence, diagnosis and treatment of alveolar echinococcosis in humans as well as possible methods of its prevention. Studies done in 2001-2003 in Poland have shown the high prevalence of the tapeworm in red foxes in the north-east (34.5%) and south-east (39.3%) of Poland with foci of infection in some counties (up to 70% foxes infected). It makes the high potential risk for human infection in these areas.

[Usefulness of the "OptiMAL Rapid Malaria test" for rapid detection of malaria imported to Poland]. / Przydatnosc zestawu "optimal rapid malaria test" do szybkiego rozpoznawania malarii importowanej do Polski.

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.

[Studies on the susceptibility of Trichomonas hominis to some abiotic factors. I. Preliminary assessment of antitrichomonal activity of selected compounds in vitro]. / Badania nad wrazliwscia Trichomonas hominis na czynniki abiotyczne. I. Wstepna ocena przezywalnosci wiciowców w wybranych mediach w warunkach in vitro.

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.

Mechanical transmission of Cryptosporidium parvum oocysts by flies.

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.

Malaria relapse and recrudescence among travellers to the tropics.

This study describes 14 cases of relapse and recrudescence of malaria, treated between 1991 and 2003. In that period, 146 patients were hospitalized in the Clinic of the Institute in Gdynia: 20 women and 126 men. In 103 cases the disease was caused by Plasmodium falciparum, in 31 cases by Plasmodium vivax, in 5 cases by Plasmodium malariae, and in 2 cases by Plasmodium ovale. Five patients were found to have mixed infections, with either P. falciparum and P. vivax or P. falciparum and P. ovale. Relapses in patients previously treated in the country or abroad accounted for 9.6% of all the treated cases of malaria. Recrudescences and relapses were diagnosed of both the tropical malaria (6 cases), and the tertian malaria caused by P. vivax (4 cases). Moreover, in 4 patients diagnosis was made of secondary malaria due to P. vivax infection, while the primary attack was caused by invasion of P. falciparum. Also discussed was the issue of drug-resistance of plasmodia and the resulting problems with the treatment.

Assessment of clinical course and outcome of Plasmodium falciparum malaria in Angola diagnosed by microscopic and molecular methods.

Malaria still remains the main infectious cause of morbidity and mortality in the world. The majority of cases occur in tropical Africa, including Angola. The study was performed in Angola, in the Health Centre of St. Lucas in Kifangondo, near Luanda during the dry season, from May to October 1999. 168 patients with symptoms of malaria were referred for study enrolment. In the study we assessed the efficacy of treatment in falciparum malaria based on clinical, microscopic and polymerase chain reaction (PCR) examination, and estimated the prevalence of mutations in the pfcrt gene in the P. falciparum isolates from chloroquine-treated patients. In 159 (95%) patients P. falciparum malaria was diagnosed using microscopic methods. This was confirmed by PCR. We admitted 68 patients to the hospital; 13 patients died from complicated malaria. We have analysed 59 cases of P. falciparum malaria known to be chloroquine-treated. In 58 (98%) cases we have found P. falciparum isolates that carried gene polymorphisms typical for chloroquine resistance. In the group with such P. falciparum isolates, 8 people were treated successfully. Reasons for this response are discussed.

Human alveolar echinococcosis in Poland: 1990-2011.

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.