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.

Investigations on the occurrence of Plasmodium knowlesi in travellers returning from the endemic areas of simian malaria.

Malaria remains an important public health issue all over the world. Among 5 Plasmodium species invasive to humans, Plasmodium knowlesi has been identified most recently. It is sometimes difficult to differentiate this species from P. malariae with the use of microscopic examination. However, P. knowlesi infection may be associated with rapidly increasing parasitaemia and severe clinical course with the risk of death. Samples from Polish travellers returning from areas where simian malaria is endemic were examined with the use of polymerase chain reaction (PCR). The small subunit of ribosomal RNA (SSU rRNA) genes was subjected to analysis using nested PCR reaction. No positive results of P. knowlesi were obtained. Due to morphological similarities to P. malariae, potentially severe clinical course of infection and P. knowlesi endemic regions being a common tourist destination, diagnostic and clinical vigilance is necessary, including molecular methods use for precise parasite identification.

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.

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.

Severe malaria--analysis of prognostic symptoms and signs in 169 patients treated in Gdynia in 1991-2005.

In the period 1991-2005, 169 patients with the diagnosis of malaria were hospitalized in the Department of Tropical and Parasitic Diseases, Institute of Maritime and Tropical Medicine in Gdynia (from 2003--the Academic Centre of Maritime and Tropical Medicine, Medical University of Gdansk). All the cases were analysed for severity, occurrence of complications and permanent sequelae of the disease. According to the criteria set by the WHO (5), malaria was classified as severe in 36 cases. All of them were Plasmodium falciparum infections or mixed infections: P. f. and another species of the parasite. Patients in this group developed a number of complications, inter alia shock, acute respiratory distress syndrome (ARDS), acute renal failure, blackwater fever, severe anemia, disseminated intravascular coagulation, myocarditis, consciousness disorders of varied degree, acute transient psychoses, and exacerbation of ischemic heart disease. In one case of a pregnant woman, necrosis of the fetus occurred in the course of disease in the 4th month of pregnancy. Moreover, meningoencephalitis was diagnosed in two patients--in one of them concurrently with symptoms and signs of malaria, while in the other one-3 weeks after the symptoms subsided. In 6 patients, permanent sequelae of the disease developed and in 4 patients the disease was fatal. The cause of death was multi-organ failure, with the first sign of poor prognosis being rapidly progressing renal failure resistant to treatment in three men; in one case death resulted from cerebral malaria. In cases of suspected malaria, relapsing malaria or in mixed infections, molecular testing was a valuable complementary tool of diagnosis, which helped in beginning the appropriate treatment.

Cytokines and clinical manifestations of malaria in adults with severe and uncomplicated disease.

Pro- and anti-inflammatory cytokines are supposed to be involved in malaria pathogenesis. Their relationship with clinical manifestations of the disease, however, is rarely studied in adults from non-endemic countries with imported disease, particularly with severe malaria. In this study we compared serum levels of gamma interferon (IFNgamma) and interleukins: IL-12, IL-18, IL-10 in healthy adults and patients with severe or uncomplicated imported malaria, with predominance of Plasmodium falciparum and Plasmodium vivax infections within studied group. Severe malaria was shown to be associated with elevated serum levels of IFNgamma and IL-18 as well as with relative deficiency of IL-12 mediated response in comparison to uncomplicated malaria cases, while IL-10 was found to be higher in all malaria patients compared to the controls. Overall, the results of our study are consistent with the observations from the regions with holoendemic malaria transmission, suggesting a pivotal role of impaired IL-12 expression in severe malaria. On the contrary, patients with severe malaria included into our study presented with the pattern of excessive production of inflammatory IFNgamma and IL-18, what seems to be an unusual finding compared to the results of the studies on African children and may be the feature of severe malaria in non-immune adults.

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.

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.

[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.

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.

High yield expression and single-step purification of Toxoplasma gondii SAG1, GRA1, and GRA7 antigens in Escherichia coli.

This report describes a simple, highly efficient and reproducible method for obtaining large quantities of highly pure recombinant Toxoplasma gondii antigens, which can be used for diagnostic application. The obtained T. gondii SAG1, GRA1, and GRA7 antigens (as fusion proteins), expressed in Escherichia coli, contained polyhistidine tags at the N- and C-ends that allowed single-step isolation by metal-affinity chromatography on Ni(2+)-IDA-Sepharose columns. The immunoreactivity of the recombinant antigens was tested in an enzyme-linked immunosorbent assay (ELISA) format for potential application in the serodiagnosis of T. gondii infection.