Surgical treatment of alveolar echinococcosis: a single centre experience and systematic review of the literature.

INTRODUCTION: Alveolar echinococcosis (AE) is a zoonosis caused by Echinococcus multilocularis. AE is primarily localised in the liver. Echinococcus multilocularis imitates tumour-like behaviour. It can metastasise through blood or lymphatic system to distant organs. Echinococcosis often remains asymptomatic due to its long incubation period and indistinct symptoms. Clinical symptoms are determined by the parasites location. Diagnosis of echinococcosis is based on medical history, clinical symptoms, laboratory tests, serology results, imaging methods and final histology findings. Surgical removal of the cyst with a safety margin, followed by chemotherapy is the therapeutic method of choice. CASE REPORT: We present a case report of alveolar echinococcosis in a thirty-year-old female patient in whom we surgically removed multiple liver foci of alveolar echinococcosis. The disease recurred after two years and required another surgical intervention. CONCLUSIONS: Alveolar echinococcosis is a disease with a high potential for a complete cure provided that it is diagnosed early and that the recommended therapeutic procedures are strictly adhered to.

Toxocara spp. seronegativity in Czech patients with early form of multiple sclerosis - clinically isolated syndrome.

BACKGROUND: Helminth infections were reported to slightly modulate the host immune system response and decrease the risk of an autoimmune disorder, but on the other hand any infection may activate the immune system and trigger autoimmune reaction. In this study, we aimed to measure eosinophil levels and antibodies against Toxocara spp. in patients with clinically isolated syndrome (CIS) and multiple sclerosis (MS). METHODS: In total, 220 CIS patients and 62 MS patients were examined. Antibodies against Toxocara secretory/excretory antigens (TES) were measured with an ELISA method. RESULTS: A total of 1,983 measurements of eosinophil levels were performed in CIS patients, out of which 95 results in 21 different patients were above the upper normal limit of the laboratory, but it was mostly only a relative increase. Two patients showed eosinophil levels above 20 % but both of them suffered from severe allergy. None of the CIS patients had any clinical signs of parasitic infections and the serological tests for antibodies against Toxocara were all negative. In all MS patients, eosinophil levels were in normal range. Antibodies against TES were detected in only 1 out of 62 (1.6%) MS patients. CONCLUSIONS: Based on our results it does not seem that Toxocara infection represents a potential trigger of MS. Nevertheless, our study indirectly confirms the hypothesis that parasitic infection may protect from autoimmunity.

[Human alveolar echinococcosis and an overview of the occurrence of Echinococcus multilocularis in animals in the Czech Republic]. / Humánní alveolární echinokokóza a prehled výskytu tasemnic Echinococcus multilocularis u zvírat v Ceské republice.

Human alveolar echinococcosis (AE) is caused by larval stages of the tapeworm Echinococcus multilocularis. In the Czech Republic, screening tests to detect the specific infectious agent have been performed since 1998. The first AE cases were diagnosed in 2007, and until 2014, a total of 21 diseases were recorded. In accordance with radiological, histological, and/or PCR data, serological examinations of 699 individuals helped to reveal 15 additional AE cases in the period of 2015-2016. From the cumulative data for 1998-2016, it appears that of 2,695 patients examined, 36 (18 men and 18 women) were diagnosed with AE. Their age at diagnosis ranged from 20 to 82 years and was lower for women (mean 43.7, median 39.5) than for men (50.9 and 57.5, respectively), but the difference was not statistically significant. In the period of 2007-2016, the mean annual incidence rate was 0.034 cases/100 000 population. Our study indicates an ongoing increase in AE cases. The disease can be autochthonous in nature, as evidenced not only by some case history data but also by the detection of the larval stages in wild boar (Sus scrofa). AE risk to humans in the Czech Republic is discussed in the context of the known data on the presence of various parasite developmental stages in animals.

The effect of a probiotic Escherichia coli strain on regulatory T-cells in six year-old children.

Probiotics are believed to prevent or reduce allergy development but the mechanism of their beneficial effect is still poorly understood. Immune characteristics of regulatory T cells (Tregs) in peripheral blood of perinatally probiotic-supplemented children of allergic mothers (51 children), non-supplemented children of allergic mothers (42 children), and non-supplemented children of healthy mothers (28 children) were compared at the age of 6-7 years. A first dose of a probiotic Escherichia coli strain (E. coli O83:K24:H31) was administered within 2 days after the birth and then 12 times during the first months of life and children were followed longitudinally. Proportion and functional properties of Tregs were estimated by flow cytometry in relation to the children's allergy status. Proportion of Tregs in the peripheral blood of children suffering from allergy tends to be higher whereas median of fluorescence intensity (MFI) of FoxP3 was significantly decreased in allergic group. Intracellular presence of regulatory cytokine interleukin (IL)-10 was also lower in allergic children. Immune functions of Tregs reflected by both MFI of FoxP3 and IL-10 in the group of probiotic-supplemented children of allergic mothers were nearly comparable with children of healthy mothers while probiotic non-supplemented children of allergic mothers have decreased immune function of Tregs. Supplementation by probiotic E. coli strain decreases allergy incidence in high-risk children. In contrast to our expectation, proportion of Tregs has not been increased in probiotic supplemented children. Beneficial effect of probiotics on newborn immature immune system could be, at least partially, explained by the modulating immune function of Tregs. In summary, we detected increased proportion of Tregs in peripheral blood of allergic children, their functional properties were decreased in comparison with the Tregs of healthy children. A unifying hypothesis for these findings is that Treg numbers in allergic children are increased in order to compensate for decreased function.

Alveolar echinococcosis - a rare disease with differential diagnostic problems.

INTRODUCTION: Alveolar echinococcosis is a life-threatening zoonotic parasitic disease. Its incidence is rare. In some cases, the correct and timely diagnosis can be difficult. CASE REPORT: The authors present the case of a young patient with liver, diaphragm and lung involvement. The suspicion of echinococcus infection was made on the basis of medical history, clinical symptoms, and a combination of ultrasonography, computed tomography, magnetic resonance imaging tests and serological methods. The patient underwent multimodal treatment with albendazole and en-bloc resection of the liver, lung and diaphragm. The definitive diagnosis of alveolar echinococcosis was determined from samples of the resected tissues using histopathology and polymerase chain reaction methods. The patient has been followed regularly and is on life-long treatment with albendazole. CONCLUSION: The precise diagnosis and multimodal therapy of alveolar echinococcosis is fundamental from the point of view of patient long-term survival. KEY WORDS: alveolar echinococcosis - diagnosis - multimodal treatment - follow-up.

Trichobilharzia anseri n. sp. (Schistosomatidae: Digenea), a new visceral species of avian schistosomes isolated from greylag goose (Anser anser L.) in Iceland and France.

Parasitological investigations carried out on birds in Iceland and France highlight the presence of four species of avian schistosomes from greylag geese (Anser anser L.): the european nasal species Trichobilharzia regenti and three visceral species, among which an unknown species isolated from blood vessels of the large intestine and liver. Morphological and molecular analyzes of different parasite stages (eggs, adults) revealed new species of Trichobilharzia genus ­ Trichobilharzia anseri sp. nov. Studies on host-parasite relationship under natural conditions, showed that the life-cycle includes the snail Radix balthica (syn. R. peregra) as intermediate host. The cercariae, already isolated in Iceland from two ponds of the Reykjavik capital area ­ the Family park and Tjörnin Lake ­ are the same as those isolated in 1999 by Kolárová et al. during the first study on Icelandic parasitic agents of cercarial dermatitis.

[Antibiotic resistance in nontyphoidal salmonellae serovars in the Czech Republic]. / Antibiotická rezistence u netyfových sérovaru Salmonella spp. v Ceské republice.

STUDY AIM: To determine antibiotic resistance and incidence of multidrug resistance among Nontyphoidal salmonellae serovars isolated from humans. MATERIAL AND METHODS: Consecutive Salmonella isolates from patients, recovered in 48 microbiology laboratories in May 2012, were analyzed in the respective reference laboratories at the National Institute of Public Health. Strains were re-identified and differentiated into serovars. Their minimum inhibitory concentrations (MICs) to 11 antibiotics were determined by the microdilution method. RESULTS: Of 25 serovars identified among 637 strains of Salmonella enterica, the most frequent were Enteritidis (87.0 %), Typhimurium (4.9 %), and monophasic Typhimurium 4,[5],12:i:- (2.0 %) and Mbandaka (0.6 %); other serovars were rare. Altogether 558 strains (87.6 %) were susceptible to all antibiotics tested and the remaining 79 strains were resistant to one or more antibiotics. The prevalence rates of resistance to individual antibiotics among 637 study strains were as follows: ampicillin 8.5%, tetracycline 5.7%, sulfamethoxazole 5.2%, cipro-floxacin 3.8%, and chloramphenicol 2.5%. Resistance to gentamicin, trimethoprim, and third and fourth generation cephalosporins was rare ( 0.5%) and none of the study strains showed resistance to meropenem. Three producers of extended spectrum beta-lactamase were multidrug resistant and two of them recovered from twins exhibited a different pattern of resistance. Resistant strains were most often assigned to the following serovars: Enteritidis (49.4%), Typhimurium (26.6%), and monophasic Typhimurium (15.2%). While only 7% (39 of 554 strains) of Enteritidis strains were resistant, the serovars Typhimurium and its monophasic variant 4,[5],12:i:- showed high rates of resistance, i.e. 66.7 and 92.3%, respectively. Furthermore, resistance was revealed in all strains of the serovars Virchow (n = 3), Kentucky (n = 1), and Newport (n = 1), in two of three strains of the serovar Infantis, and in one of two strains of the serovar Stanley. All five blood isolates were assigned to the serovar Enteritidis and one of them showed resistance to ciprofloxacin. Of 79 resistant strains, 26.6% showed resistance to ampicillin only and 24.1% to ciprofloxacin only, with multidrug resistance, i.e. resistance to three or more antibiotics, confirmed in 43.0% of strains. CONCLUSION: Despite a relatively low prevalence of resistance to the antibiotics tested among 637 study strains, the following alarming findings were made: Detection of Salmonella enterica strains resistant to ciprofloxacin as the drug of choice or to higher generation cephalosporins and multidrug resistance revealed in two thirds of the strains of the serovar Typhimurium and in all but one strains of its monophasic variant 4,[5],12:i:-.

Molecular diversity of Trichobilharzia franki in two intermediate hosts (Radix auricularia and Radix peregra): a complex of species.

Recently, the systematic use of the molecular approach as a complement to the other approaches (morphology, biology, life cycle) has brought help for the identification of species considered as different in the past to be regrouped and synonymised, and distinctions to be drawn between species similar at the morphological level. Among these species, we tried to clarify the situation of Trichobilharzia frankiMüller and Kimmig, 1994, species that today include more than 50 haplotypes notably coming from larval stages isolated from intermediate hosts belonging to gastropods of the Radix genus. Cercariae were isolated in France and Iceland from various molluscs, before being analyzed, with their hosts, by molecular analysis of various fields such as the D2 and ITS of the ribosomal DNA and the COX1 of mitochondrial DNA. We thus show the presence of two clades depending on the specificity of their intermediate host in which they were isolated (Radix auricularia or Radix peregra), thus allowing separation of the species T. franki that had been described in the past as a probable new species.

Methodical approaches in the identification of areas with a potential risk of infection by bird schistosomes causing cercarial dermatitis.

Larval stages (cercariae) of schistosomatid flukes represent the causative agents of swimmer's itch (cercarial dermatitis), a waterborne allergic disease. Cercariae of bird schistosomes are the most frequently reported agent. Recent studies on parasite behaviour in mammals showed that infections by cercariae can be linked to more than skin syndromes. Despite the failure of complete development in mammals, bird schistosomes can escape from the skin and migrate transitorily in the hosts. These findings brought novel insights into the fate and potential pathogenic effect of the parasites in non-compatible hosts, including humans. Cercarial dermatitis occurs globally and recently is considered to be re-emerging; however, there are no data on the number of afflicted persons per year. This might be explained by a relatively low interest in human skin infections arising after bathing in fresh water. In addition, the real occurrence of bird schistosomes in the field is known only for a few areas. The paucity of epidemiological/biogeographical data is probably caused by difficulties associated with detection of the parasites in intermediate and definitive hosts. Therefore, based on personal experience and data available in the literature, we have summarized methodological approaches enabling the detection of bird schistosomes in various hosts and environments.

[Alveolar echinococcosis--a new emerging disease?]. / Alveolární echinokokóza--nove se objevující onemocnení?

Alveolar echinococcosis is a rare zoonotic disease caused by the cestode larval stages of Echinococcus multilocularis. The Larva develops as a tumour like cyst (hydatid). Due to a slow cyst development, the early stage of the infection is always asymptomatic and the clinical symptoms develop after years later. In the Czech Republic the disease is still very rare. Only one alveolar echinococcosis case was reported 30 years ago. However, new alveolar echinococcosis cases occurred during last two years and some of them indicate autochtonic character of the transmission. With regard to the recent increasing risk of infection with E. multilocularis in Central Europe, we present two case reports of alveolar echinococcosis in a 33-year old woman and a 24-year old man. The reports show diagnostic process and therapy of the infection.

A review on swimmer's itch and the occurrence of bird schistosomes in Iceland.

In the past decade, swimmer's itch (SI) has repeatedly occurred in people who have been wading or bathing in ponds or lakes in Iceland where water birds and snails are abundant. Some of the affected sites were warmed by geothermal activity, and others were not. A search for the causative agent of SI, ocellate furcocercariae that have been found in Iceland only in Radix peregra snails, revealed an average infection prevalence of 1.4% (n = 12,432). Locally, infection rates commonly exceeded 6%, the highest value observed being 24.5%. A search for adult schistosomes in visceral organs and the nasal cavities of 110 water birds belonging to the orders Gaviiformes, Podicipediformes and Anseriformes revealed eggs, miracidia or adult stages of at least seven previously identifiable schistosome species in four anseriform bird species. A previously unknown species of schistosome, Allobilharzia visceralis, was detected in whooper swans (Cygnus cygnus), and classified in a new genus. In mallards (Anas platyrhynchos) a nasal Trichobilharzia sp. and the visceral schistosome T. franki were identified. In red-breasted merganser (Mergus serrator), distinct egg types belonging to two species of the genus Trichobilharzia have been found. In grey-lag goose (Anser anser) two different egg types were also found - a large Trichobilharzia sp. and small eggs of a Dendritobilharzia sp. Additionally, unidentified cercariae, probably belonging to a previously undescribed genus were detected in R. peregra in Oslandsstjörn. Taken together, the data obtained by morphological examination of eggs and recent DNA sequencing results, indicate that at least eight species of bird schistosomes occur in Iceland.

Bird schistosome diversity in Iceland: molecular evidence.

During the years 2002-2007, Icelandic freshwater snails and birds from different orders were examined for bird schistosomes. Only the snail Radix peregra and anatid birds proved to be infected. In total, 32 samples of bird schistosome cercariae from seven localities and four samples of adults of Anas platyrhynchos and Mergus serrator from two localities were used for sequencing of the internal transcibed spacer (ITS) region of rDNA. Based on the sequence and position in the phylogenetic tree, five species/genotypes of bird schistosomes were identified. Three of them correspond to the sequences in GenBank for Trichobilharzia franki, Trichobilharzia sp. JR-2007 from Anas penelope (isolates Pl7 and Pl10) and schistosomatid sp. JR-2004 from Physa fontinalis. The other two species/genotypes belong to the genus Trichobilharzia, but their further determination was not possible due to lack of data. All of these species/genotypes develop in R. peregra, probably the only suitable intermediate host species in Iceland. As T. franki and schistosomatid sp. JR-2004 were previously reported from other snail species, the findings from Iceland proved that, under some circumstances, bird schistosomes are able to adapt to a different snail species; some of them can even adapt to a snail belonging to a different gastropod family. High diversity of bird schistosomes in Iceland is probably linked with migratory flyways of birds, as several bird species from Iceland overwinter in western Europe as well as in North America.

Antibody responses induced by Trichobilharzia regenti antigens in murine and human hosts exhibiting cercarial dermatitis.

Cercariae of bird schistosomes (genus Trichobilharzia) are able to penetrate the skin of mammals (noncompatible hosts), including humans, and cause a Th2-associated inflammatory cutaneous reaction termed cercarial dermatitis. The present study measured the antibody reactivity and antigen specificity of sera obtained after experimental infection of mice and natural infection of humans. Sera from mice re-infected with T. regenti showed a bias towards the development of antigen-specific IgM and IgG1 antibodies and elevated levels of total serum IgE, indicative of a Th2 polarized immune response. We also demonstrate that cercariae are a source of antigens triggering IL-4 release from basophils collected from healthy human volunteers. Analysis of sera from patients with a history of cercarial dermatitis revealed elevated levels of cercarial-specific IgG, particularly for samples collected from adults (> 14 years old) compared with children (8-14 years old), although elevated levels of antigen-specific IgE were not detected. In terms of antigen recognition, IgG and IgE antibodies in the sera of both mice and humans preferentially bound an antigen of 34 kDa. The 34 kDa molecule was present in both homogenate of cercariae, as well as cercarial excretory/secretory products, and we speculate it may represent a major immunogen initiating the Th2-immune response associated with cercarial dermatitis.

Penetration of Trichobilharzia cercariae into mammals: dangerous or negligible event?

Bird schistosomes and cases of human cercarial dermatitis occur worldwide, but the number of cases is not monitored. Experiments with two schistosomes, namely Trichobilharzia szidati and T. regenti, show that they possess potent tools to penetration bird and mammalian skin, as well as exhibit species-specific migration patterns within vertebrate bodies. Therefore, the infections may affect different organs/tissues e.g. lungs or spinal cord. In this minireview, the adaptations and pathogenic effects of bird schistosomes in experimental mammals are discussed, and some ideas/hypotheses on risks to humans from exposure to bird schistosome cercariae are expressed.

High-performance liquid chromatography-tandem mass spectrometry in the identification and determination of phase I and phase II drug metabolites.

Applications of tandem mass spectrometry (MS/MS) techniques coupled with high-performance liquid chromatography (HPLC) in the identification and determination of phase I and phase II drug metabolites are reviewed with an emphasis on recent papers published predominantly within the last 6 years (2002-2007) reporting the employment of atmospheric pressure ionization techniques as the most promising approach for a sensitive detection, positive identification and quantitation of metabolites in complex biological matrices. This review is devoted to in vitro and in vivo drug biotransformation in humans and animals. The first step preceding an HPLC-MS bioanalysis consists in the choice of suitable sample preparation procedures (biomatrix sampling, homogenization, internal standard addition, deproteination, centrifugation, extraction). The subsequent step is the right optimization of chromatographic conditions providing the required separation selectivity, analysis time and also good compatibility with the MS detection. This is usually not accessible without the employment of the parent drug and synthesized or isolated chemical standards of expected phase I and sometimes also phase II metabolites. The incorporation of additional detectors (photodiode-array UV, fluorescence, polarimetric and others) between the HPLC and MS instruments can result in valuable analytical information supplementing MS results. The relation among the structural changes caused by metabolic reactions and corresponding shifts in the retention behavior in reversed-phase systems is discussed as supporting information for identification of the metabolite. The first and basic step in the interpretation of mass spectra is always the molecular weight (MW) determination based on the presence of protonated molecules [M+H](+) and sometimes adducts with ammonium or alkali-metal ions, observed in the positive-ion full-scan mass spectra. The MW determination can be confirmed by the [M-H](-) ion for metabolites providing a signal in negative-ion mass spectra. MS/MS is a worthy tool for further structural characterization because of the occurrence of characteristic fragment ions, either MS( n ) analysis for studying the fragmentation patterns using trap-based analyzers or high mass accuracy measurements for elemental composition determination using time of flight based or Fourier transform mass analyzers. The correlation between typical functional groups found in phase I and phase II drug metabolites and corresponding neutral losses is generalized and illustrated for selected examples. The choice of a suitable ionization technique and polarity mode in relation to the metabolite structure is discussed as well.

Diversity of bird schistosomes in anseriform birds in Iceland based on egg measurements and egg morphology.

Radix balthica is the only freshwater mollusc in Iceland that has been found to be the intermediate host of ocellate furcocercariae. Infections are common. On average, 4.8% out of almost 7,000 snails examined during 2003-2006 were found to shed cercariae in late summer and autumn. Search for adult bird schistosome worms, eggs and miracidia in the intestines and in the nasal cavity of 110 water birds of the orders Gaviiformes (one species), Podicipediformes (one species) and Anseriformes (nine species) revealed 39 (35.5%) infected hosts. Exclusively, anseriform birds were infected. Based on morphological features and measurements of eggs, seven distinct bird schistosome species were detected in four of the bird species. Allobilharzia visceralis was found in 26.7% of the whooper swans Cygnus cygnus. In Anas platyrhynchos, the nasal Trichobilharzia sp. I (prevalence 73.3%) and the visceral Trichobilharzia sp. III (66.7%) were detected. In Mergus serrator, the distinct egg types of Trichobilharzia sp. IV and Trichobilharzia sp. V were found in 83.3% of examined birds. In Anser anser, also two different egg types were found: large eggs, referred to as Trichobilharzia sp. II (prevalence 54.2%), and the small eggs of Dendritobilharzia sp. (8.3%). Egg size and shape comparisons indicate that Trichobilharzia sp. III and T. franki might be the same species.

[Ocular sparganosis in the Czech Republic--a case report]. / Ocní sparganóza v Ceské republice: Kazuistika.

The authors report a case of 14-year-old boy presented to the Ophthalmologic Department of University Hospital in Olomouc with the diagnosis af acute anterior uveitis. A living parasite has been detected in the anterior chamber in a slit lamp examination. The vitreous and retina remained uninvolved. The acute iridocyclitis associated with parasitic infection is a very rare cause of anterior uveitis in Central Europe. The patient underwent surgical removal of the parasite via paracentesis. The inflammation resolved under the treatment with atropine and dexamethasone drops within a few days. Histopathological examination revealed the parasite as a young larval stage of tapeworm from family Pseudophyllidea (sparganum). Based on the analysis of development cycles of different types of tapeworms and according the literature data on tapeworms found in the Czech Republic genus Spirometra seemed to be the most plausible cause of the patient's disease. As the most probable source of viable parasites authors suspect swallowed water containing tiny infected crustaceans in the dam near the town Olomouc in Moravia.

[Schistosomiasis cases at the Teaching Hospital Královské Vinohrady in Prague]. / Prípady schistosomózy ve Fakultní nemocnici Královské Vinohrady.

In our paper we present the diagnosis of schistosomiasis at the Teaching Hospital Královské Vinohrady in Prague. This helminthiasis is imported to the Czech Republic by tourists and foreigners from subtropical and tropical regions of Africa, Asia, Central and South America. An unrecognized and thus untreated infection may have serious, even fatal consequences for the patient. In the period 2003 to 2005 in our parasitological investigations we have seen two cases infested with two fluke species imported from Africa. We submit the case histories of these patients.

[Hydatic cysts--echinococcosis]. / Echinokoková nákaza--echinococcosis.

The case diagnosis and treatment of hydatic cyst in the right and left lobe of the liver in a 28 year old pregnant woman is described in the article.

Molluscan and vertebrate immune responses to bird schistosomes.

There is a growing understanding of risks posed by human contact with the cercariae of bird schistosomes. In general, there are no fundamental biological differences between human and bird schistosomes in terms of their interactions with snail and vertebrate hosts. The penetration of host surfaces is accompanied by the release of penetration gland products and the shedding of highly antigenic surface components (miracidial ciliated plates and cercarial glycocalyx) which trigger host immune reactions. New surface structures are formed during transformation: the tegument of mother sporocysts and the tegumental double membrane of schistosomula. These surfaces apparently serve as protection against the host immune response. Certain parasite excretory-secretory products may contribute to immunosuppression or, on the other hand, stimulation of host immune reactions. Discovery of new species and their life cycles, the characterization of host-parasite interactions (including at the molecular level), the determination of parasite pathogenicity towards the host, the development of tools for differential diagnosis and the application of protective measures are all topical research streams of the future. Regularly updated information on bird schistosomes and cercarial dermatitis can be found at http://www.schistosomes.cz (web pages of Schistosome Group Prague).