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 Impact of Short-Term, Intensive Antifolate Treatment (with Pyrimethamine and Sulfadoxine) and Antibiotics Followed by Long-Term, Secondary Antifolate Prophylaxis on the Rate of Toxoplasmic Retinochoroiditis Recurrence.

PURPOSE: To assess the impact of intensive antifolate treatment, followed by secondary antifolate prophylaxis (A-SP) on the recurrence rate of toxoplasmic retinochoroiditis (TRC). To investigate whether there are any other factors potentially predisposing for recurrence. MATERIAL AND METHODS: A total of 637 medical records of TRC patients, who had been treated in the years 1994-2013 were reviewed. All patients were treated with pyrimethamine /sulfadoxine one 25mg/500mg tablet daily (P/S 25/500mg) for 21 days with a double loading dose for the first two days. From Day 2 the patients also received prednisone at a starting dose of 40mg and spiramycine 3 million IU three times daily, given for 10 days followed by azithromycin 500mg once daily for another 6 days. The analysis of the recurrence rate involved 352 patients who had completed 6-month secondary prophylaxis (P/S one 25 mg/500mg tablet twice a week). RESULTS: When secondary antifolate prophylaxis (A-SP) was instituted immediately after the treatment for TRC, the probability of 3-year recurrence-free survival after the first course of A-SP was 90.9%. A recurrence was most likely approximately 3.5 years after the first treatment. A univariate Cox regression model demonstrated that a risk for recurrence was 2.82 times higher (p = 0.02) in patients with retinal scars. In the multivariate analysis, the risk for recurrence was 2.41 higher (p = 0.06). In patients with haemorrhagic lesions the risk for recurrence was lower, aRR = 0.17 (approaching borderline statistical significance p = 0.08). CONCLUSIONS: With the institution of A-SP of immediately after the intensive treatment for TRC, i.e. when a reactivation was most likely, there was no recurrence during A-SP. Following A-SP the recurrence rates were low and recurrence-free periods tended to be longer. The treatment regimen employed had a beneficial effect on the recurrence interval as it reduced and delayed the highest probability of recurrence.

Symptomatic co-infection with Babesia microti and Borrelia burgdorferi in patient after international exposure; a challenging case in Poland.

The report presents a well-documented case of symptomatic co-infection of Babesia microti and Borrelia burgdorferi in a Polish immunocompetent patient after travelling to Canada and the USA.

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.

Risk of human toxocarosis in Poland due to Toxocara infection of dogs and cats.

The purpose of this paper was to present the current data on the risk of toxocarosis in humans in Poland and to give an overview of the clinical and diagnostic aspects of the disease. The number of reported clinical cases of Toxocara infection in children in Poland in medical literature has increased recently. The results of field surveys aimed to evaluate the soil contamination with geohelminth eggs conducted during the last few years showed that Toxocara is the most common zoonotic agent in urban public sites and in rural settlements. The questionnaire revealed rural inhabitants' low awareness of zoonotic parasite threats to humans. In particular parents should be advised as to what proper preventive measures to undertake to eliminate the toxocarosis risk factors for children in rural environment. Prevention of initial environment contamination with Toxocara canis and T. cati eggs, which includes proper treatment regimes to eliminate patent infections in dogs and cats and preventing pets from defecating in public areas and private households is vital. To provide the public with suitably presented information as well as pet owners with uniform recommendations, a close collaboration between veterinary and public health professionals is crucial.

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.

[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.

Tularaemia after tick exposure - typical presentation of rare disease misdiagnosed as atypical presentation of common diseases: a case report.

A 44-year-old female was admitted because of tender, enlarged inguinal lymph nodes with a history of tick bite five weeks earlier. In the place of a tick bite on the skin a small ulcer was present. The primary symptoms before admission suggested typical diseases related to tick bite such as Lyme borreliosis and tick-borne encephalitis, what corresponded with positive IgM ELISA test for Lyme borreliosis. The course of disease however clarified the diagnosis of tularaemia, which is a relatively rare disease in Poland (6 cases per 40 million population are reported annually). The ultimate diagnosis was confirmed by serological tests.

[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.

[Toxocariasis in children--difficult clinical problem]. / Toksokaroza u dzieci--trudny problem kliniczny.

Toxocariasis in humans is a zoonosis due to the migration of Toxocara canis or T. cati larvae in human body. This review provides basic information on pathology of infection and clinical signs and symptoms of toxocariasis in children. The cases diagnosed in the recent years in central Poland are presented. The disease was recognised accidentally based on the results of laboratory analysis performed for other reasons, when abnormalities suggesting an active helminthiasis were found (eosinophilia, elevated IgE level and/or anemia). The high rate of soil contamination in households of the patients demonstrated elevated risk of infection and reinfection in both, rural and urban areas. Presented data show the need for educational programs which should be implemented for prevention of Toxocara infection in children.

[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.