Publikacje

Immunoproteomic analysis of Trichinella spiralis and Trichinella britovi excretorysecretory muscle larvae proteins recognized by sera from humans infected with Trichinella

Sylwia GrzelakID, Anna StachyraID, Jerzy Stefaniak2, Karolina Mrówka2, Bożena MoskwaID, Justyna Bień-KalinowskaID

Abstract
The present study compares the immunogenic patterns of muscle larvae excretory-secretory proteins (ML E-S) from T. spiralis and T. britovi recognized by Trichinella-infected human sera. Samples were analyzed using two-dimensional electrophoresis (2-DE) coupled with 2D-immunoblot and liquid chromatography-tandem mass spectrometry LC-MS/
MS analysis, two ELISA procedures and a confirmatory 1D-immunoblot test. Sera were obtained from nine patients with a history of ingestion of raw or undercooked meat who presented typical clinical manifestations of trichinellosis and from eleven healthy people. Specific anti-Trichinella IgG antibodies were detected in all samples tested with the Home- ELISA kits, but in only four samples for the commercially-available kit. The 1D-immunoblot results indicated that all nine serum samples were positive for T. spiralis ML E-S antigens, expressed as the presence of specific bands. In contrast, eight of the serum samples with T. britovi E-S ML antigens were positive, with one serum sample taken from a patient at 33dpi (days post infection) being negative. To identify immunoreactive proteins that are specifically recognized by host antibodies, both species of ML E-S proteins were subjected to 2Dimmunoblotting with human serum taken at 49 dpi. The sera recognized 22 protein spots for T. spiralis and 18 for T. britovi in 2D-immunoblot analysis. Their molecular weights (MW) ranged from 50 to 60 kDa. LC-MS/MS analysis identified both common and specifically-recognized immunoreactive proteins: transmembrane serine protease 9, serine protease, antigen targeted by protective antibodies and Actin-1 partial were shared for both Trichinella species; hypothetical protein T01_7775 and P49 antigen, partial those specific to T. spiralis; deoxyribonuclease-2-alpha and hypothetical protein T03_17187/T12_7360 were specific to T. britovi. Our results demonstrate the value of 2-DE and 2D-immunblot as versatile tools for pinpointing factors contributing to the parasite-host relationship by comparing the secretomes of different Trichinella species.

The Seroprevalence of Toxoplasma gondii in Wild Boars from Three Voivodeships in Poland, MAT Analyses

Aleksandra Kornacka, Bożena Moskwa, Anna Werner, Piotr Nowosad, Wiesława Jankowska, Aleksandra Cybulska, Anna C. Majewska

Abstract
Purpose
The European wild boar (Sus scrofa) is a popular game animal species. Its meat, however, can represent a reservoir
of dangerous foodborne diseases and can play an important role in the transmission of many pathogens, including Toxoplasma gondii, in humans and animals worldwide. The aim of the present study was to determine the presence of antibodies to T. gondii in the serum of hunted wild boars in Poland.
Methods
Using the commercial direct agglutination test, 398 serum samples collected during the hunting season 2009/2010
were tested for the presence of T. gondii antibodies, and the titre of 40 was considered indicative of T. gondii infection in analysed samples.
Results
It was found that nationwide, 37.7% were seropositive to T. gondii, although seroprevalence varied from 11.6 to 50%
depending on the Voivodeship. Significant differences were observed between the Great Poland and Lubusz Voivodeships and between Great Poland and Warmian-Masurian.
Conclusion
Serological test indicated widespread exposure to T. gondii by wild boar; therefore, consumption of raw or  undercooked game meat of infected animals can carry a significant risk of T. gondii infection. Keywords Toxoplasma gondii · Wild boars · Modified agglutination test · Antibodies

Summary
Strongyloidiasis and hookworm infections are neglected helminth diseases widespread in tropical and subtropical areas. In humans, particularly in immunocompromised patients infections potentially may lead to the life-threatening clinical conditions involving the respiratory as well as gastrointestinal systems. The increased number of tourists travelling to tropical regions is associated with more frequent infection with parasites such as Strongyloides and hookworm. The infection takes place when fi lariform larvae penetrate the skin exposed to soil, than migrate through the lungs and fi nally reach the intestine. Travelers are often not aware of how they could get infected. Physicians may suspect strongyloidiasis and hookworm infections in tourists with diarrhea returning from endemic areas, especially when an elevated eosinophilia is observed. In the literature there are many reports about the presence of parasites in indigenous communities, but very few are available regarding travelers. This paper describes a dual infection with hookworm and Strongyloides stercoralis in a young female tourist returning from Southeast Asia. To our knowledge, this is the fi rst report of hookworm and Strongyloides stercoralis infection in a tourist from Europe, acquired in an endemic area.
Keywords: Strongyloides; hookworms; soil-transmitted helminths; tourists; diarrhea

Molecular Epidemiology and Genetic Diversity of Orthohantaviruses in Small Mammals in Western Poland

Seung-Ho Lee, Jin Sun No, Won-Keun Kim, Ewa Gajda, Agnieszka Perec-Matysiak, Jeong-Ah Kim, Joanna Hildebrand,
Richard Yanagihara and Jin-Won Song

Abstract
Orthohantaviruses are negative-sense, single-stranded RNA viruses harbored by multiple small mammals.
Dobrava-Belgrade virus (DOBV) and Puumala virus (PUUV) cause hemorrhagic fever with renal syndrome (HFRS) in
Europe. In Poland, serological surveys have demonstrated antibodies against DOBV and PUUV in patients with HFRS.
Molecular evidence of DOBV and PUUV has been found in Apodemus flavicollis and Myodes glareolus, respectively, in
southeastern Poland, and Seewis virus (SWSV) has been reported in Sorex araneus in central Poland. However, data on
the geographic distribution and phylogeny of orthohantaviruses are unavailable for other regions in Poland. To ascertain the prevalence and genetic diversity of orthohantaviruses in western and northern Poland, lung tissues from 106 small mammals were analyzed for the presence of orthohantavirus RNA. DOBV and SWSV were detected in two of 42 (4.8%) Apodemus agrarius and in three of 10 (30%) S. araneus, respectively. Phylogenetic analyses of partial L- and S-segment sequences of DOBV indicated a shared genetic lineage with the Kurkino genotype from Slovakia, Russia, and Hungary, whereas the partialMsegment ofDOBVclustered with the Kurkino genotype from Germany. Phylogenetic relationships of the SWSV L and S segments showed a geographic lineage with SWSV strains from central Poland, Czech Republic, and Germany. In conclusion, the study provides insights into the molecular prevalence, phylogenetic diversity, and evolutionary relationship ofDOBVin A. agrarius andSWSVin S. araneus. This report increases awarenessamong physicians for HFRS outbreaks in western Poland.

The usefulness of commercially available serological tests in the diagnosis and monitoring of treatment in patients with alveolar echinococcosis

Małgorzata Sulima, Beata Szostakowska, Wacław Nahorski, Katarzyna Sikorska, Wojciech Wołyniec, Piotr Wąż

Abstract
Aim of the study: To assess the clinical usefulness of serological tests in the diagnosis and monitoring of treatment
of patients with alveolar echinococcosis (AE).
Material and methods: The results of serological tests, i.e. Echinococcus multilocularis ELISA (Bordier Affinity
Products) and Echinococcus Western Blot IgG (LDBIO Diagnostic), of 66 patients were analysed. Duration of
follow-up was two years after diagnosis. In the second phase of the study 11 sera obtained from the patients
undergoing surgical treatment, in whom the results of Echinococcus Western Blot IgG assay were still positive,
were additionally tested with Anti-Echinococcus EUROLINE-WB (IgG) assay.
Results: Statistically significant negativization of the Echinococcus multilocularis ELISA test was observed in the
group of patients who underwent radical surgery or liver transplantation. Negativization of Echinococcus Western
Blot IgG assay results was observed in some patients, among both those who received conservative treatment and
those who underwent surgery, but no statistically significant differences were found between treatment groups.
In 54.5% of cases the Anti-Echinococcus EUROLINE-WB (IgG) test result was negative when the results of the
Echinococcus Western Blot IgG assay were still positive.
Conclusions: Echinococcus multilocularis ELISA proved to be useful in assessing the activity of AE in a group of
patients who underwent radical surgery or liver transplantation. The results of our study suggest that Anti-Echinococcus EUROLINE-WB (IgG) is a more dynamic test, which at the time of disappearance of AE activity becomes negative earlier.
Key words: serology, echinococcosis, alveolar echinococcosis.

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Polskie Towarzystwo Parazytologiczne
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