Advanced imaging of microorganisms, including protists, is challenging due to their small size. Specimen expansion prior to imaging is thus beneficial to increase resolution and cellular details. Here, we present a sample preparation workflow for improved observations of the single-celled eukaryotic pathogen Giardia intestinalis (Excavata, Metamonada). The binucleated trophozoites colonize the small intestine of humans and animals and cause a diarrhoeal disease. Their remarkable morphology includes two nuclei and a pronounced microtubular cytoskeleton enabling cell motility, attachment and proliferation. By use of expansion and confocal microscopy, we resolved in a great detail subcellular structures and organelles of the parasite cell. The acquired spatial resolution enabled novel observations of centrin localization at Giardia basal bodies. Interestingly, non-luminal centrin localization between the Giardia basal bodies was observed, which is an atypical eukaryotic arrangement. Our protocol includes antibody staining and can be used for the localization of epitope-tagged proteins, as well as for differential organelle labelling by amino reactive esters. This fast and simple technique is suitable for routine use without a superresolution microscopy equipment.

• Klíčová slova
• Caltractin, Centrin, Cytoskeleton, Expansion microscopy, Giardia, Protist,
• MeSH
• Giardia lamblia * ultrastruktura   cytologie   MeSH
• konfokální mikroskopie * MeSH
• lidé MeSH
• organely ultrastruktura   chemie   MeSH
• protozoální proteiny analýza   chemie   MeSH
• trofozoiti ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• protozoální proteiny MeSH

Out of three genotypes of Encephalitozoon cuniculi (I-III) available for experimental studies, E. cuniculi genotype I remains the less characterized. This study describes for the first time individual phases of microsporidiosis caused by E. cuniculi genotype I and efficacy of albendazole treatment in immunocompetent BALB/c and C57Bl/6 mice and immunodeficient SCID, CD4-/- and CD8-/- mice using molecular detection and quantification methods. We demonstrate asymptomatic infection despite an intense dissemination of microsporidia into most organs within the first weeks post infection, followed by a chronic infection characterized by significant microsporidia persistence in immunocompetent, CD4-/- and CD8-/- mice and a lethal outcome for SCID mice. Albendazole application led to loss E. cuniculi genotype I infection in immunocompetent mouse strains, decreased spore burden by half in CD4-/- and CD8-/- mice, and prolongation of survival of SCID mice. These results showed Encephalitozoon cuniculi genotype I infection extend and albendazole sensitivity was comparable to E. cuniculi genotype II, but the infection onset speed and mortality rate was similar to E. cuniculi genotype III. These imply that differences in the course of infection and the response to treatment depend not only on immunological status of the host, but also on the genotype causing the infection.

• Klíčová slova
• Encephalitozoon cuniculi genotype I, Experimental infection, Laboratory mice, RT PCR quantification,
• MeSH
• albendazol aplikace a dávkování   MeSH
• antigeny CD4 genetika   MeSH
• antigeny CD8 genetika   MeSH
• antiinfekční látky aplikace a dávkování   MeSH
• Encephalitozoon cuniculi klasifikace   genetika   MeSH
• encephalitozoonóza imunologie   parazitologie   MeSH
• genotyp MeSH
• imunokompetence MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši SCID MeSH
• myši MeSH
• polymerázová řetězová reakce MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• albendazol MeSH
• antigeny CD4 MeSH
• antigeny CD8 MeSH
• antiinfekční látky MeSH

Non-invasive small animal in vivo imaging is an essential tool in a broad variety of biomedical sciences and enables continuous monitoring of disease progression in order to develop and improve diagnostic, therapeutic and preventive measures. Imaging parasites non-invasively in live animals allows efficient parasite distribution evaluation in the host organism and objective evaluation of parasitic diseases' burden and progression in individual animals. The aim of this systematic review was to summarize recent trends in small animal in vivo imaging and compare and discuss imaging of single-cell and multicellular eukaryotic parasites. A literature survey was performed using Web of Science and PubMed databases in research articles published between 1990 and 2018. The inclusion criteria were using any imaging method to visualize a range of protozoan and helminth parasites in laboratory animals in vivo. A total of 92 studies met our inclusion criteria. Protozoans and helminths were imaged in 88% and 12% of 92 studies, respectively. The most common parasite genus studied was the protozoan Plasmodium followed by Trypanosoma and Leishmania. The most frequent imaging method was bioluminescence. Among the helminths, Schistosoma and Echinococcus were the most studied organisms. In vivo imaging is applicable in both protozoans and helminths. In helminths, however, the use of in vivo imaging methods is limited to some extent. Imaging parasites in small animal models is a powerful tool in preclinical research aiming to develop novel therapeutic and preventive strategies for parasitic diseases of interest both in human and veterinary medicine.

• Klíčová slova
• Bioluminescence, Fluorescence, Helminth, Luciferase, Mouse, Protozoan,
• MeSH
• hlodavci parazitologie   MeSH
• králíci parazitologie   MeSH
• luminiscenční měření metody   MeSH
• parazitární nemoci u zvířat diagnostické zobrazování   MeSH
• zvířata MeSH
• Check Tag
• králíci parazitologie   MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• systematický přehled MeSH

Silymarin (SIL) represents a natural mixture of polyphenols showing an array of health benefits. The present study, carried out on a model cestode infection induced by Mesocestoides vogae tetrathyridia in the ICR strain of mice, was aimed at investigating the impact of SIL as adjunct therapy on the activity of praziquantel (PZQ) in relation to parasite burden, immunity and liver fibrosis within 20 days post-therapy. In comparison with PZQ alone, co-administration of SIL and PZQ stimulated production of total IgG antibodies to somatic and excretory-secretory antigens of metacestodes and modified the expression patterns of immunogenic molecules in both antigenic preparations. The combined therapy resulted in the elevation of IFN-γ and a decline of TNF-α and TGF-β1 in serum as compared to untreated group; however, SIL attenuated significantly the effect of PZQ on IL-4 and stimulated PZQ-suppressed phagocytosis of peritoneal macrophages. In the liver, SIL boosted the effect of PZQ on gene expression of the same cytokines in a similar way as was found in serum, except for down-regulation of PZQ-stimulated TNF-α. Compared to PZQ therapy, the infiltration of mast cells into liver after SIL co-administration was nearly abolished and correlated with suppressed activities of genes for collagen I, collagen III and α-SMA. In conclusion, co-administration of SIL modified the effects of PZQ therapy on antigenic stimulation of the immune system and modulated Th1/Th2/Tregs cytokines. In liver this was accompanied by reduced fibrosis, which correlated with significantly higher reduction of total numbers of tetrathyridia after combined therapy as compared with PZQ treatment.

• Klíčová slova
• Fibrosis, Immunity, Mesocestoides vogae, Parasite counts, Praziquantel, Silymarin,
• MeSH
• antioxidancia aplikace a dávkování   MeSH
• cestodózy farmakoterapie   MeSH
• cytokiny účinky léků   MeSH
• down regulace účinky léků   MeSH
• Mesocestoides účinky léků   MeSH
• myši inbrední ICR MeSH
• myši MeSH
• praziquantel aplikace a dávkování   MeSH
• silymarin aplikace a dávkování   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antioxidancia MeSH
• cytokiny MeSH
• praziquantel MeSH
• silymarin MeSH

The availability of high quality genomic DNA in sufficient amounts to perform Next Generation Sequencing (NGS) experiments is challenging for pathogens that cannot be cultivated in vitro, as is the case for many parasites. Therefore, Whole Genome Amplification (WGA) of genomic DNA is used to overcome this limitation. In this study, we evaluated the effect of WGA using the intestinal flagellated protozoan Giardia duodenalis as a model, due to its genome compactness (12 Mb), the presence of two diploid nuclei with variable levels of allelic sequence heterogeneity (ASH), and the availability of reference genomes. We selected one isolate (ZX15) belonging to the same genetic group of the reference isolate WB, namely Assemblage A, sub-Assemblage AI. Genomic DNA from the ZX15 isolate (GEN dataset) and that obtained by WGA of 1 ng of the same genomic DNA (WGA dataset) were sequenced on a HiSeq Illumina platform. Trimmed reads from the GEN and WGA experiments were mapped against the WB reference genome, showing the presence of a very small number of mutations (846 and 752, respectively). The difference in the number of mutations is largely accounted by local variation in coverage and not by bias introduced by WGA. No significant difference were observed in the distribution of mutations in coding and non-coding regions, in the proportion of heterozygous mutations (ASH), or in the transition/transversion ratio of Single Nucleotide Variants within coding sequences. We conclude that the quantitative and qualitative impact of WGA on the identification of mutations is limited, and that this technique can be used to conduct comparative genomics studies.

• Klíčová slova
• Assemblage AI, Comparative genomics, Giardia duodenalis, Mutations, Next generation sequencing, Whole genome amplification,
• MeSH
• celogenomová asociační studie MeSH
• Giardia lamblia genetika   MeSH
• giardiáza parazitologie   MeSH
• lidé MeSH
• mutace MeSH
• otevřené čtecí rámce genetika   MeSH
• předškolní dítě MeSH
• protozoální DNA chemie   genetika   izolace a purifikace   MeSH
• strukturální variace genomu MeSH
• techniky amplifikace nukleových kyselin MeSH
• výpočetní biologie MeSH
• Check Tag
• lidé MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• protozoální DNA MeSH

The use of medicinal plants (MP) containing bioactive compounds is an alternative strategy to control of parasitic nematode of small ruminants Haemonchus contortus at various stages of their life cycle. The aims of this study were to determine the in vitro anthelmintic activity of both aqueous and methanolic extracts from 13 medicinal plants typical for Central Europe, and to determine quantity of selected plant secondary metabolites (PSMs) in the methanolic extracts. In vitro egg hatch test and larval development tests were conducted to determine the possible anthelmintic effects of methanolic and aqueous extracts of the roots of Althaea officinalis L., Petasites hybridus L. and Inula helenium L.; flowers of Malva sylvestris L. and Chamomilla recutita L.; leaves of Plantago lanceolata L. and Rosmarinus officinalis L.; seeds of Foeniculum vulgare Mill. and stems of Solidago virgaurea L., Fumaria officinalis L., Hyssopus officinalis L., Melisa officinalis L. and Artemisia absinthium L. on eggs and larvae of H. contortus. Ultra-performance liquid chromatography and tandem mass spectroscopy was used for quantifying six PSMs: gallic acid (GA), rutin (RU), diosmin (DI), hesperidin (HE), quercetin (QU) and kaempferol (KA). RU content of the most effective methanolic extracts was in the order: M. sylvestris (9.33 mg/g DM) > A. absinthium (6.10 mg/g DM) > C. recutita (0.42 mg/g DM). The highest concentration of QU (44.8 mg/g DM) and KA (6.59 mg/g DM) were detected in stems of F. officinalis comparing to the other evaluated plants. The most significant (p < 0.05) anthelmintic effects exhibited methanolic extracts of A. absinthium in both in vitro tests (i.e., egg hatch test and larval development test). Additionally, only two methanolic extracts of C. recutita and M. sylvestris were comparable to activity of A. absinthium using the larval development test. Wider spectrum of aqueous extracts exhibited stronger ovicidal activity in comparison to methanolic extracts. The similar trend was observed in evaluating of larvicidal activity of aqueous and methanolic plant extracts.

• Klíčová slova
• Anthelmintic activity, Bioactive compounds, Haemonchus contortus, In vitro tests, UPLC/MS/MS,
• MeSH
• anthelmintika izolace a purifikace   farmakologie   MeSH
• Artemisia absinthium chemie   MeSH
• chromatografie kapalinová MeSH
• feces parazitologie   MeSH
• Fumaria chemie   MeSH
• Haemonchus účinky léků   růst a vývoj   MeSH
• kempferoly analýza   farmakologie   MeSH
• larva účinky léků   MeSH
• léčivé rostliny chemie   MeSH
• Malva chemie   MeSH
• Matricaria chemie   MeSH
• ovce MeSH
• ovum účinky léků   MeSH
• quercetin analýza   farmakologie   MeSH
• rostlinné extrakty farmakologie   MeSH
• tandemová hmotnostní spektrometrie MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH
• Názvy látek
• anthelmintika MeSH
• kaempferol MeSH Prohlížeč
• kempferoly MeSH
• quercetin MeSH
• rostlinné extrakty MeSH

Microsporidia are obligate intracellurar unicellular parasite of wide range of vertebrates. Although ingestion or inhalation of microsporidian spores is the main route of infection, assumed vertical transmission was described in some mammals. The present study was focused on proof of vertical transmission in mice under experimental conditions. Mice were infected with E. cuniculi genotype II intraperitoneally after mating, or perorally followed by mating in acute or chronic phase of infection. Fetuses were delivered by Caesarean section or mice were kept up to the parturition. Some of cubs were immediately after birth transferred to uninfected surrogate mothers. Group of cubs was immunosuppressed. All cubs were examined using polymerase chain reaction for the presence of Encephalitozoon after birth or in their age of 3 or 6 weeks, respectively. All fetuses delivered by Caesarean section, which were intraperitoneally or perorally infected were negative as well as all neonatal mice and youngsters tested in age of 6 weeks. Only immunosuppressed cubs and cubs of immunodeficient mice in age of 21 days were positive for Encephalitozoon cuniculi genotype II. Present results provided the evidence that transplacental transmission of Encephalitozoon cuniculi in mice occurs, but the mechanism of these transport is still unknown.

• Klíčová slova
• BABL/c, Encephalitozoon cuniculi, Microsporidia, SCID, Transplacental transmission,
• MeSH
• Cercopithecus aethiops MeSH
• DNA fungální chemie   izolace a purifikace   MeSH
• Encephalitozoon cuniculi klasifikace   genetika   MeSH
• encephalitozoonóza imunologie   mikrobiologie   přenos   MeSH
• genotyp MeSH
• hostitel s imunodeficiencí MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední BALB C MeSH
• myši SCID MeSH
• myši MeSH
• spory hub MeSH
• těhotenství MeSH
• Vero buňky MeSH
• vertikální přenos infekce * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA fungální MeSH

Microsporidia are a group of obligate intracellular eukaryotic parasites, which are able to infect a wide range of animals, including humans. Four genotypes of Encephalitozoon cuniculi have been found to date. The different courses of microsporidiosis described in humans, which are dependent on immunological status of the host and genotype of E. cuniculi, have been successfully imitated in murine models. In the present study, we quantified the microsporidial burden in individual organs of a murine experimental model, using qPCR and we compared the parasitic load of two genotypes of E. cuniculi, namely genotype II and III (EC II and EC III). While the extent of microsporidiosis caused by EC II gradually increased over 35 days post infection (DPI) in both immunocompetent and immunodeficient mice and caused death in the latter at 28 DPI, EC III had spread into all host organs by seven DPI and was not lethal for either mouse strain during the experimental time period. Moreover, EC III persisted in many organs until termination of the experiment. The number of microsporidial spores in individual organs was ten times higher in EC III-infected animals compared to those infected with EC II. EC II infection also progressively shifted towards organs outside the gastrointestinal tract (GIT) in both monitored mouse strains; whereas, EC III infection equally remained in both the GIT and organs outside the GIT. With the increasing use of molecular methods in diagnostics, it is important to better understand the pathophysiology of microsporidia, including its ability to escape from the immune system and persist in host organisms. Our results indicate that pathogenicity is not directly connected to spore burden, as infection caused by E. cuniculi genotype II is less extensive and spreads more slowly within the host organism than infection caused by E. cuniculi genotype III, but which caused the earlier death of immunodeficient mice.

• Klíčová slova
• BALB/C, Encephalitozoon cuniculi genotypes II and III, Microsporidiosis, SCID, qRT-PCR,
• MeSH
• Arvicolinae MeSH
• Cercopithecus aethiops MeSH
• Encephalitozoon cuniculi klasifikace   genetika   růst a vývoj   fyziologie   MeSH
• encephalitozoonóza parazitologie   MeSH
• gastrointestinální trakt parazitologie   MeSH
• genotyp MeSH
• imunokompetence MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• Microsporidia fyziologie   MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední BALB C MeSH
• myši SCID MeSH
• myši MeSH
• parazitární zátěž MeSH
• spory hub MeSH
• Vero buňky MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Blastocystis is a common inhabitant of the human gut, colonizing at least one billion people at a prevalence ranging from <10% to 100% in healthy human populations globally. The majority of carriers remain asymptomatic, suggesting that Blastocystis is largely a commensal, though Blastocystis has also been implicated in disease in some people. However, there are no in vivo model systems in which to experimentally test the impact of Blastocystis on mammalian hosts and the gut ecosystem and determine which factors underlie these variable clinical outcomes. We evaluated a rat model for sustaining of a human-derived Blastocystis ST1 and assess colonization success and longevity. Because of the broad host range of Blastocystis, we compared the rat with three other rodent species to establish the reproducibility of our method. Blastocystis was introduced by esophageal gavage and colonization success evaluated by Blastocystis culture. Culture was also used to determine that all animals were negative prior to colonization and negative controls remain Blastocystis-free. In this study, Blastocystis ST1 established in 100% of the outbred rats (Rattus norvegicus) and gerbils (Meriones unguiculatus) challenged. Rats were colonized asymptomatically for more than one year, but Blastocystis ST1 was not transmitted between rats. Mus musculus strain CD1 and Mastomys coucha were not susceptible to Blastocystis ST1. Thus, rats appear to be a suitable in vivo model for studies of Blastocystis ST1, as do gerbils though testing was less extensive. This work lays the foundation for experimental work on the role of Blastocystis in health and disease.

• Klíčová slova
• Blastocystis ST1, Colonization longevity, Experimental colonization, In vivo model, Rat, Susceptibility to colonization,
• MeSH
• Blastocystis růst a vývoj   patogenita   MeSH
• blastocystóza diagnóza   parazitologie   MeSH
• centrifugace - gradient hustoty MeSH
• feces parazitologie   MeSH
• Gerbillinae MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• modely nemocí na zvířatech * MeSH
• Murinae MeSH
• myši MeSH
• náchylnost k nemoci MeSH
• organismy bez specifických patogenů MeSH
• potkani Wistar MeSH
• zdravotní stav MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• srovnávací studie MeSH

Host- and age-specificity of Cryptosporidium avium were studied in 1-, 21- and 365-day-old chickens (Gallus gallus), domestic ducks (Anas platyrhynchos) and ring-necked pheasants (Phasianus colchicus) under experimental conditions. Cryptosporidium avium was not infectious for ring-necked pheasants, but it was infectious for ducks and chickens at all age categories. The course of infection in ducks did not differ among age categories, but 365-day-old chickens had less severe infections than 1- and 21-day-old chickens. The patent period in chickens and ducks was >30 DPI, but ducks started to shed oocysts of C. avium earlier (5-6 DPI) and at a lower intensity (accumulated value of infection intensity of 58,000-65,000 OPG) than chickens (9-11 DPI and accumulated value of infection intensity of 100,000-105,000 OPG). Experimentally infected birds showed no clinical signs of cryptosporidiosis.

• Klíčová slova
• Anseriformes, Course of infection, Cryptosporidium avium, Galliformes, Transmission study,
• MeSH
• Cryptosporidium klasifikace   genetika   izolace a purifikace   MeSH
• divoká zvířata MeSH
• feces parazitologie   MeSH
• Galliformes parazitologie   MeSH
• gastrointestinální obsah parazitologie   MeSH
• genotypizační techniky MeSH
• hospodářská zvířata MeSH
• hostitelská specificita MeSH
• kachny parazitologie   MeSH
• kryptosporidióza imunologie   parazitologie   MeSH
• kur domácí parazitologie   MeSH
• nemoci drůbeže imunologie   parazitologie   MeSH
• nemoci ptáků imunologie   parazitologie   MeSH
• odolnost vůči nemocem MeSH
• oocysty izolace a purifikace   MeSH
• polymerázová řetězová reakce veterinární   MeSH
• protozoální DNA chemie   genetika   izolace a purifikace   MeSH
• RNA ribozomální chemie   genetika   MeSH
• věkové faktory MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• protozoální DNA MeSH
• RNA ribozomální MeSH