Cystic echinococcosis in Poland: genetic variability and the first record of Echinococcus granulosus sensu stricto (G1 genotype) in the country.

Cystic echinococcosis is one of the most important zoonotic diseases affecting humans and livestock worldwide, and is endemic in Poland. A set of six isolates on larval stages of Echinococcus granulosus sensu lato tapeworms collected from three humans, two pigs and one sheep from Polish foci of CE was examined by DNA sequencing of two mitochondrial genes (cox1, rrnS). The results demonstrated the presence of E. canadensis and E. granulosus sensu stricto in the investigated hydatid cysts. The former species was found in all five isolates from pigs and humans derived from central Poland. In a sheep hydatid cyst originating from Lesser Poland Voivodeship in southern Poland, E. granulosus s. s. (G1 genotype) was identified. This is the first report of an unambiguously autochthonous infection with E. granulosus s. s. in Poland. The global distribution and host affiliations of the commonly occurring G1 microvariant with nucleotide change 56C/T in cox1, detected here in Polish sheep, are discussed. The finding that sheep harboured E. granulosus s. s. may have important consequences for developing effective hydatid control programmes in Poland due to its longer maturation rate in dogs compared with E. canadensis G7. This may lead to greater expenditures for purchasing anthelmintics to provide an appropriate dosing regime in sheep-raising areas of the country.

Genetic diversity of Blastocystis hominis sensu lato isolated from humans in Poland

INTRODUCTION AND OBJECTIVES: Blastocystis hominis s. l. is one of the most commonly detected protozoa in the human large intestine. The aim of the study was to determine the genetic subtypes of Blastocystis hominis s. l. occurring in humans in Poland. MATERIALS AND METHODS: Stool samples from patients diagnosed in the Laboratory of the Department of Parasitology, National Institute of Public Health ­ National Institute of Hygiene (NIZP-PZH) and in the Parasitology Laboratory of the Hospital for Infectious Diseases in Warsaw were examined. Blastocystis subtypes were assayed based on the fragment of small-subunit ribosomal RNA gene sequences (SSU rDNA). RESULTS: The examined isolates were classified into five Blastocystis subtypes (STs), fifteen of which belonged to ST3, three to ST1, two to ST2, two to ST6, and one isolate belonged to ST7. In three cases the subtype of isolate was not identified. DISCUSSION AND CONCLUSIONS: In Poland, the subtypes ST1, ST2, ST3, ST4, ST6 and ST7 have been reported in humans so far. The ST6 and ST7 subtypes are rarely detected in humans in Europe. In Poland, the ST6 subtype was previously described in chickens. On the basis of the studies, it was found that Blastocystis isolated from humans in Warsaw show high genetic diversity. In order to determine the possible pathogenic potential of individual Blastocystis subtypes, special epidemiological studies are required.

The first case of human autochtonous subconjunctival dirofilariosis in Poland and MALT lymphoma as possible consequence of this parasitosis.

The first case of human dirofilarosis in Poland was recorded in 2007. Until that time our country was free of Dirofilaria repens. Recent studies show that 21,4- 60% of dogs in Warsaw region harbour microfilariae, therefore it is becoming a growing problem in Central Europe. In April 2013 a subconjunctival D. repens was removed from the eye of 61-year-old woman. It was the twenty first case of this disease in Poland, the third case of eye dirofilaria and the fourth autochtonous case. The patient had never been abroad, so it was the first case of autochtonous human ocular dirofilariosis in Poland. Nine months after the D. repens had been removed, a MALT lymphoma was discovered. In the article we discuss whether a MALT lymphoma of the lacrimal gland of the eye, previously affected by the parasite, may be the consequence of the invasion.

Human dirofilariasis due to Dirofilaria repens in Ukraine, an emergent zoonosis: epidemiological report of 1465 cases.

The filarial nematode Dirofilaria repens is currently considered to be one of the most extensively spreading human and animal parasites in Europe. In Ukraine, reporting cases of dirofilariasis has been mandatory since 1975, and the disease was included in the national surveillance system for notifiable diseases. Up until December 31st 2012, a total of 1533 cases have been registered, with 1465 cases occurring within the previous 16 years. Most of the cases of dirofilariasis were registered in 6 regions: Kyiv, and the Donetsk, Zaporizhzhya, Dnipropetrovsk, Kherson and Chernihiv oblasts. In the years 1997-2002 the highest incidence rate was noted in the Kherson oblast in the south of the country (9.79 per 100 000 people), and the lowest in western Ukraine (0.07-1.68 per 100 000 people). D. repens infections were registered in all oblasts. Parasitic lesions were most often located in the head, the subconjunctival tissue and around the eyes. D. repens lesions were also found in the limbs, torso, male sexual organs, and female mammary glands. Dirofilariasis was diagnosed in persons aged from 11 months to 90 years old, most often among people between 21-40 years of age. Most patients had only one parasitic skin lesion; the majority of isolated nematodes were female. The results of our analysis point to a constant increase in D. repens dirofilariasis incidence in humans in Ukraine. Despite educational efforts, infections have become more frequent and the territory in which the disease occurs has enlarged to encompass the whole of Ukraine. Nevertheless, the Ukrainian sanitary-epidemiological services managed to achieve some measure of success, e.g. by creating a registration system for D. repens infections and establishing proper diagnostics for the disease.

Vector-borne helminths of dogs and humans - focus on central and eastern parts of Europe.

Comments on the article "Vector-borne helminths of dogs and humans in Europe" published in Parasites and Vectors 2013, 6:16.

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.

Human dirofilariosis in Poland: the first cases of autochthonous infections with Dirofilaria repens.

Dirofilaria (Nochtiella) repens Railliet et Henry, 1911 (Nematoda: Onchocercidae) is a subcutaneous parasite of dogs and other carnivorous animals, with human acting as incidental hosts. D. repens occurs endemically in warm climates on various continents, in Europe mainly in Mediterranean countries. The aim of this study was to summarize information on human dirofilariosis in Poland, taking into consideration parasitological and epidemiological data. Between April 2009-December 2011, in the parasitological laboratories of Medical University in Warsaw and the National Institute of Public Health/National Institute of Hygiene, fragments of affected human tissues and parasite specimens were examined microscopically. Molecular methods were used to confirm the results from eight microscopic investigations. A literature review to summarize all data on dirofilarial infections in humans in Poland was conducted. In these investigations, autochthonous dirofilariosis was found in humans for the first time in Poland. During the last 3 years, 12 new cases of human D. repens dirofilariosis were recognized. Since 2007, a total of 18 D. repens infection have been found in humans in Poland. Parasitic changes were located in various parts of the body, in the form of subcutaneous nodules containing single nematodes surrounded by granulation tissue (15 cases). In 3 cases, a subconjuctival localization was found. Seventeen of the 18 described cases were noted in central Poland where dirofilariosis occured in dogs. In this area, autochtonous infection was identified in 3 women who had never left Poland in their lives; the others were probably infected outside the country while staying in endemic regions. Data on human and canine infection collected from central Poland during the last 5 years indicates that Dirofilaria repens has been introduced into our country, and that the infection is successfully spreading, with the border of the endemic area currently on 52°N, 21°E. To control the epidemiological situation it is necessary to identify D. repens hosts within local mosquito populations, and to monitor dogs. Because of the increasing number of cases of human infections, whether introduced or local, physicians should take dirofilariosis into consideration in differential diagnosis of skin and eye diseases.

The Polish collection of parasitic helminths (a report on realization of works concerning fusion of parasitic collections dispersed among different scientific institutions).

The paper presents the results of works on preparation of a proper museum collection of parasitic helminths gathered by the Polish scientists and dispersed in various scientific institutions. The collection composed of 11 author's collections and a special collection of available typical series of species described by the Polish authors has been deposited in the Museum of Natural History of Wroclaw University. It includes almost 16.000 slides with 90 taxons of Monogenea, 251 Trematoda, 144 Cestoda, 43 Nematoda, 11 Acanthocephala, in total 486 nominal species and 53 taxons of genus level. Among them 12 species appear new for fauna of Poland; for 57 species new hosts in Polish territory have been recorded; two new species of Cestoda have been described in separate publications. A special paragraph contains a checklist of all species described by the Polish scientists with information whether they are available in any other collection. Detailed information about every deposited slide is given in the Database available online at http://www.helminths.eu .

[Procedures for species identification of the nematodes belonging to Dirofilaria genus present in the clinical material isolated from humans]. / Rozpoznawanie izolowanych od ludzi nicieni z rodzaju dirofilaria przy zastosowaniu technik biologii molekularnej.

Procedures for DNA extraction and amplification were modified to allow identification of Dirofilaria nematodes surgically removed from human tissues. Worm samples stored in: ethanol (24 weeks), formalin (46 weeks) or paraffin blocks (25weeks) were examined. Fragments of two ribosomal DNA regions (5.8S-ITS2-28S, 5SrRNA) and mitochondrial cytochrome oxidase subunit I gene were used as diagnostic markers. The highest PCR sensitivity was observed for DNA obtained from the worm preserved in ethanol, while the nucleic acid extracted form the parasite stored in formalin yielded the lowest PCR sensitivity. DNA extraction from the parasite preserved in formalin was more time consuming than DNA extraction from the remaining samples. Furthermore, the amplification of DNA isolated from the formaldehyde preserved worm allowed for identification of the parasite species only when the mitochondrial marker was used in Real Time PCR, and the amount of the obtained product was close to the detection limit. Species identification of the worms stored in the paraffin block and in ethanol was possible with both traditional and Real Time PCR. All analyzed worms were identified as D. repens which confirmed the species identification based on morphological features. The results show that molecular methods are relatively simple to use and suitable for identification ofDirofilaria sp. nematodes present in clinical material. Formalin is not suitable for storing material intended for molecular tests.

[Usage of the Hoyer's medium for diagnostics and morphological studies of some parasites]. / Zastosowanie plynu Hoyera do diagnostyki i badan morfologicznych niektórych pasozytów.

Various modification of the mixture of gum arabic with chloral hydrate can be used for mounting of small arthropods as well as some helminths. However, in diagnostic laboratories in Poland they seem to remain unknown. The authors of current work present examples of the Hoyer's medium application. The medium has been composed according to the initial, given by Hoyer, hundred-years-old recipe, which was the root of all other, later used recipes. Hoyer's medium is universal in action and very comfortable to use in slides for microscope investigation. It gives the immediate light-through effect, so it can be helpful in fast diagnostics. At the same time it allows to store the slides for a relatively long time e.g. with education purpose.

[Dirofilariasis--Dirofilaria repens--first time described in Polish patients]. / Dirofilarioza-Dirofilaria repens--po raz pierwszy opisana u polskich pacjentów.

In this paper the first cases of human dirofilariasis in Poland are presented. The clinical manifestations were typical subcutaneous nodules with the adult worm (D. repens) in the center. Past medical history of the 5 patients revealed their trips to endemic countries, so the imported infection was considered. However 2 of these cases were difficult to interpret. It is unclear whether the infections should be considered endemic or imported, because of patients' short visits abroad which happened long time (10 years) ago.

[Molecular diagnosis of oral cavity trichomonas infections in HIV patients]. / Diagnostyka molekularna rzesistków jamy ustnej u pacjentów z HIV.

The occurrence of trichomonas in oral cavity of HIV patients is not well known. HIV patients often suffer from oral lesions (candidiosis, advanced caries) and it remains unclear if any oral parasites can affect that, therefore the aim of the study was verification of species that can occur in HIV patients' oral cavity. Diagnosis of oral trichomonas can be performed by conventional methods (microscopic observation of wet and stained preparations and cultivation) but these are time consuming and insufficient for proper species differentiation, therefore in order to detect and identify species of parasites precisely, molecular methods such as PCR (Polymerase Chain Reaction) and sequencing of its product, were applied. 54 HIV patients (18 females and 36 males at the age of 20-54) were examined. All of them were addicted to intravenous drugs for at least 6 years. Saliva, smears and spittle samples were collected and used for cultivation, preparations and molecular diagnosis. For PCR amplification a pair of primers (T1 and T2) specific for ITS 1 - 5.8 SrRNA - ITS 2 region was designed. The oral trichomonas were detected in saliva samples of 3 HIV patients; these were males at the age of 25, 27 and 44. The identification of species by PCR and sequencing of the PCR products showed the trichomonads belonging to Trichomonas tenax. Infection of HIV patients' oral cavity caused by T. tenax is rather related with inflammatory processes than with the immunosuppression of these patients but should be considered as a potential factor in pathogenesis of oral disorders in immunosuppressed patients.

[Prevalence of intestinal nematodes in dogs from Warsaw region]. / Wystepowanie nicieni jelitowych u psów w okolicach Warszawy.

Prevalence of intestinal nematodes in dogs from Warsaw region. Investigation of prevalence of intestinal parasites in dogs have been conducted in order to protect human and animal health. The aim of the study was to establish species composition of intestinal parasites and to evaluate their prevalence in dogs from the shelter in Józefów situated on north from Warsaw. Additionally, urban dogs from Warsaw and village dogs from areas near the shelter, were examined. The prevalence of nematodes was: 62.3% in dogs from shelter, 37.5% in village dogs and 18.8% in urban dogs. In homeless dogs, the most common parasite was Uncinaria stenocephala, then Trichuris vulpis and Toxascaris leonina. In village dogs only eggs of U. stenocephala were detected; in urban dogs Toxocara canis and U. stenocephala were found.