INTRODUCTION: Hammondia hammondi and Toxoplasma gondii are closely related protozoan parasites, but only T. gondii is zoonotic. Both species use felids as definitive hosts and cannot be differentiated by oocyst morphology. In T. gondii, a 529-base pair (bp) repetitive element (TgREP-529) is of utmost diagnostic importance for polymerase chain reaction (PCR) diagnostic tests. We identified a similar repetitive region in the H. hammondi genome (HhamREP-529). METHODS: Based on reported sequences, primers and probes were selected in silico and optimal primer probe combinations were explored, also by including previously published primers. The analytical sensitivity was tested using serial dilutions of oocyst DNA. For testing analytical specificity, DNA isolated from several related species was used as controls. The newly established TaqMan PCR (Hham-qPCR1) was applied to tissues collected from H. hammondi-infected gamma-interferon gene knockout (GKO) mice at varying time points post-infection. RESULTS: Ten forward and six reverse primers were tested in varying combinations. Four potentially suitable dual-labelled probes were selected. One set based on the primer pair (Hham275F, Hham81R) and the probe (Hham222P) yielded optimal results. In addition to excellent analytic specificity, the assay revealed an analytical sensitivity of genome equivalents of less than one oocyst. Investigation of the tissue distribution in GKO mice revealed the presence of parasite DNA in all examined organs, but to a varying extent, suggesting 100- to 10,000-fold differences in parasitic loads between tissues in the chronic state of infection, 42 days post-infection. DISCUSSION: The use of the 529-bp repeat of H. hammondi is suitable for establishing a quantitative real-time PCR assay, because this repeat probably exists about 200 times in the genome of a single organism, like its counterpart in T. gondii. Although there were enough sequence data available, only a few of the primers predicted in silico revealed sufficient amplification; the identification of a suitable probe was also difficult. This is in accord with our previous observations on considerable variability in the 529-bp repetitive element of H. hammondi. CONCLUSIONS: The H. hammondi real-time PCR represents an important novel diagnostic tool for epidemiological and cell biological studies on H. hammondi and related parasites.

• MeSH
• diferenciální diagnóza MeSH
• feces parazitologie   MeSH
• kočky parazitologie   MeSH
• kokcidióza veterinární   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• molekulární patologie metody   MeSH
• myši parazitologie   MeSH
• oocysty genetika   izolace a purifikace   MeSH
• protozoální geny MeSH
• repetitivní sekvence nukleových kyselin MeSH
• Sarcocystidae * genetika   izolace a purifikace   MeSH
• Toxoplasma * genetika   izolace a purifikace   MeSH
• toxoplazmóza zvířat MeSH
• zvířata MeSH
• Check Tag
• kočky parazitologie   MeSH
• myši parazitologie   MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In the original article [1], the authors Dr Jasmin OMERAGIĆ and Dr Davor ALAGIĆ were erroneously omitted from the co-authors list.

• Publikační typ
• časopisecké články MeSH
• tisková chyba MeSH

BACKGROUND: Taenia solium and Taenia saginata are food-borne parasites of global importance. In eastern Europe only fragmented information is available on the epidemiology of these zoonotic parasites in humans and animal populations. In particular for T. solium, on-going transmission is suspected. The aim of this systematic review was to collect the available data and describe the current knowledge on the epidemiology of T. solium and T. saginata in eastern Europe. METHODS: Literature published in international databases from 1990 to 2017 was systematically reviewed. Furthermore, local sources and unpublished data from national databases were retrieved from local eastern European experts. The study area included 22 countries. RESULTS: Researchers from 18 out of the 22 countries provided data from local and unpublished sources, while no contacts could be established with researchers from Belarus, Kosovo, Malta and Ukraine. Taeniosis and human cysticercosis cases were reported in 14 and 15 out of the 22 countries, respectively. Estonia, the Former Yugoslav Republic of Macedonia, Lithuania, Moldova, Poland, Romania, Serbia, and Slovakia reported cases of porcine cysticercosis. Croatia, Czech Republic, Estonia, Former Yugoslav Republic of Macedonia, Moldova, Poland, Romania, Serbia, Slovakia, and Ukraine reported bovine cysticercosis. CONCLUSIONS: There is indication that taeniosis and cysticercosis are present across eastern Europe but information on the occurrence of T. solium and T. saginata across the region remains incomplete. Available data are scarce and species identification is in most cases absent. Given the public health impact of T. solium and the potential economic and trade implications due to T. saginata, notification of taeniosis and human cysticercosis should be implemented and surveillance and notification systems in animals should be improved.

• MeSH
• cysticerkóza epidemiologie   parazitologie   MeSH
• lidé MeSH
• nemoci prasat epidemiologie   parazitologie   MeSH
• nemoci skotu epidemiologie   parazitologie   MeSH
• neurocysticercosis epidemiologie   parazitologie   MeSH
• prasata parazitologie   MeSH
• prevalence MeSH
• skot MeSH
• Taenia saginata fyziologie   MeSH
• Taenia solium fyziologie   MeSH
• tenióza epidemiologie   parazitologie   MeSH
• veřejné zdravotnictví MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Geografické názvy
• východní Evropa epidemiologie   MeSH

Undercooked lamb and mutton are common sources of Toxoplasma gondii infection for humans. A sequence specific magnetic capture technique in combination with quantitative real-time PCR targeting the 529 bp repeat element of T. gondii was used for estimation of the parasite burdens in various sheep tissues (n = 6) three months after peroral experimental inoculation with 10,000 T. gondii oocysts. Brain was the most frequently affected organ (positive in all 6 sheep) and showed the highest estimated parasite loads (0.5-30,913 parasites/g tissue). Lung samples were positive in three sheep, with load estimates of 36.3 to <1 parasite/g tissue. Heart tissue was positive in three sheep and kidney only in one animal with low parasite loads (<1 parasite/g tissue). Only few skeletal muscle samples in 2 animals showed positive results, with very low parasite burdens, while samples from further internal organs (i.e. liver and spleen) were negative in all animals. This study identified the brain as the most important predilection site and therefore the most appropriate tissue for T. gondii detection.

• MeSH
• anatomické struktury zvířat parazitologie   MeSH
• kvantitativní polymerázová řetězová reakce přístrojové vybavení   metody   MeSH
• magnetismus MeSH
• ovce MeSH
• Toxoplasma genetika   izolace a purifikace   MeSH
• toxoplazmóza zvířat parazitologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• autoimunitní nemoci parazitologie   terapie   MeSH
• cizopasní červi imunologie   MeSH
• Crohnova nemoc parazitologie   terapie   MeSH
• helmintoterapie * metody   využití   MeSH
• hlístice imunologie   MeSH
• klinická studie jako téma MeSH
• lidé MeSH
• nádory parazitologie   terapie   MeSH
• paraziti imunologie   MeSH
• randomizované kontrolované studie jako téma MeSH
• Trichinella imunologie   MeSH
• Trichuris imunologie   MeSH
• Check Tag
• lidé MeSH

Ingestion of raw or undercooked meat is a potential source of human toxoplasmosis. The aim of this study was to determine the viability of Toxoplasma gondii cysts in vacuum packed (VP) goat meat and in dry fermented sausages (DFS), and evaluate certain physical and chemical parameters, like water activity (aw), pH value, content of salt, dry matter and fat. A portion of muscle tissue from experimentally infected animals was used for production of VP meat with or without addition of 2.5% curing salt, and stored at 4 °C or at -20 °C. Results of bioassay showed that, samples of vacuum packed Toxoplasma positive meat without salt addition were alive after six weeks at 4 °C. Incubation at -20 °C supported the viability after 3 h, but not after 4 h. After 7 days in 2.5% of curing salt, samples of T. gondii VP goat meat were still viable, but not after 14 days at 4 °C. All the DFS samples were not positive for infective cysts which mean that, they do not pose a risk of T. gondii transmission. These data suggest that vacuum packaging increases the survival of T. gondii cysts.

• MeSH
• kosterní svaly parazitologie   MeSH
• kozy MeSH
• lidé MeSH
• masné výrobky analýza   parazitologie   MeSH
• maso analýza   parazitologie   MeSH
• nemoci koz parazitologie   MeSH
• obaly potravin MeSH
• spotřebitelská bezpečnost produktů MeSH
• Toxoplasma genetika   růst a vývoj   izolace a purifikace   MeSH
• toxoplazmóza zvířat parazitologie   MeSH
• vakuum MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Pigs represent an important source of food in many countries, and undercooked pork containing tissue cysts is one of the most common sources of Toxoplasma gondii infection for humans. A magnetic capture method for the isolation of T. gondii DNA and quantitative real-time PCR targeting the 529 bp TOXO repeat element were used to estimate the parasite burden in different tissues of pigs experimentally infected with T. gondii oocysts, and to determine the predilection sites of T. gondii in this host species. The highest concentration of T. gondii DNA was found in brain tissues, equivalent to [median] 553.7 (range 3857.7-121.9) parasites per gram, followed by lungs, heart and dorsal muscles with median values corresponding to 0.3 (range 61.3-0.02); 2.6 (range 7.34-0.37) and 0.6 (range 2.81-0.31) parasites per gram of tissue, respectively. Skeletal muscles from fore and hindlimb, liver and kidney presented very low infection burdens equivalent to [median] ≤0.2 parasites per gram of tissues, and no parasite DNA could be detected in the spleen. This study contributes to understanding the value of different pig tissues as a source of T. gondii infection for humans and shows that the brain, while not being of major importance as human food source, may represent a first-line selection tissue when performing non-serological surveys (e.g. bioassays, histopathological, immunohistochemical or molecular studies) to detect T. gondii infections in pigs.

• MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• magnetismus metody   MeSH
• mozek parazitologie   patologie   MeSH
• nemoci prasat parazitologie   patologie   MeSH
• prasata MeSH
• Toxoplasma chemie   genetika   izolace a purifikace   fyziologie   MeSH
• toxoplazmóza zvířat parazitologie   patologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The breeding of domestic rabbit (Oryctolagus cuniculus) for human consumption has a long tradition mainly in European and Asian countries. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animals health. This study aimed to collect sera from rabbits bred in different conditions and test the presence of Toxoplasma gondii and Encephalitozoon cuniculi antibodies. Whether infections were active or latent was assessed by determining the occurrence of IgM or IgM together with IgG antibodies which indicated active infection whereas latent infection was characterized by finding IgG antibodies only. An ELISA test was performed with 1883 sera samples collected throughout the Czech and Slovak Republics. The seroprevalence of T. gondii in 902 samples from 6 commercial farms (CF) was very low with only 4 rabbits (0.4%) being positive. In total 99 (10.1%) individuals out of 981 samples from 29 household farms (HF) were positive for T. gondii antibodies. Only 2 (50%) of the T. gondii positive CF rabbits had active infections while the rest were latently infected. The serological results showed that 35 (35.4%) rabbits from the T. gondii positive HF group suffered from active infection. Out of CF samples 185 (20.5%) were positive for E. cuniculi. Antibodies of E. cuniculi were detected in 497 (50.7%) HF rabbits. Active E. cuniculi infections were determined in 85.9% of CF and 56.3% of HF rabbits; respectively. Interestingly, the E. cuniculi positive rabbits were significantly more often positive for anti-T. gondii antibodies in comparison to E. cuniculi negative individuals. Prevalence of T. gondii in CF rabbits was negligible. According to our results meat of HF rabbits still poses a risk of T. gondii infection. Nevertheless, the risk is on its lowest level in 20 years which is apparently caused due to changes in feeding practices. The occurrence of E. cuniculi antibodies was significantly lower in rabbits from commercial farms, apparently because of better hygiene conditions.

• MeSH
• ELISA veterinární   MeSH
• Encephalitozoon cuniculi imunologie   izolace a purifikace   MeSH
• encephalitozoonóza veterinární   MeSH
• králíci imunologie   parazitologie   MeSH
• prevalence MeSH
• protilátky protozoální krev   MeSH
• séroepidemiologické studie MeSH
• Toxoplasma imunologie   izolace a purifikace   MeSH
• toxoplazmóza zvířat epidemiologie   MeSH
• veřejné zdravotnictví MeSH
• zvířata MeSH
• Check Tag
• králíci imunologie   parazitologie   MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Slovenská republika MeSH

This study focuses on mapping the life cycle of Cryptosporidium muris in two laboratory rodents; BALB/c mice and the southern multimammate rat Mastomys coucha, differing in their prepatent and patent periods. Both rodents were simultaneously experimentally inoculated with viable oocysts of C. muris (strain TS03). Animals were dissected and screened for the presence of the parasite using a combined morphological approach and nested PCR (SSU rRNA) at different times after inoculation. The occurrence of first developmental stages of C. muris in stomach was detected at 2.5 days post-infection (dpi). The presence of Type II merogony, appearing 36 h later than Type I merogony, was confirmed in both rodents. Oocysts exhibiting different size and thickness of their wall were observed from 5 dpi onwards in stomachs of both host models. The early phase of parasitization in BALB/c mice progressed rapidly, with a prepatent period of 7.5-10 days; whereas in M. coucha, the developmental stages of C. muris were first observed 12 h later in comparison with BALB/c mice and prepatent period was longer (18-21 days). Similarly, the patent periods of BALB/c mice and M. coucha differed considerably, i.e. 10-15 days vs chronic infection throughout the life of the host, respectively.

• MeSH
• Cryptosporidium růst a vývoj   fyziologie   MeSH
• druhová specificita MeSH
• feces parazitologie   MeSH
• kryptosporidióza parazitologie   patologie   MeSH
• modely nemocí na zvířatech MeSH
• Murinae MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• oocysty MeSH
• stadia vývoje * MeSH
• trofozoiti MeSH
• žaludeční sliznice patologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Undercooked meat containing tissue cysts is one of the most common sources of Toxoplasma gondii infection in humans. Goats are very susceptible to clinical toxoplasmosis, and especially kids are common food animals, thereby representing a risk for human infection. A sequence-specific magnetic capture method was used for isolation of T. gondii DNA from tissue samples from experimentally infected goat-kids and real-time PCR for the 529 bp repeat element allowed quantification of T. gondii DNA. The contamination level in different types of tissue and in two groups of goats euthanized 30 and 90 dpi was compared. The highest concentration of T. gondii DNA in both groups of goats was found in lung tissue, but only the higher parasite count in lung tissue compared to other organs in group A (euthanized 30 dpi) was statistically significant. T. gondii concentrations were higher in liver and dorsal muscle samples from goats euthanized 90 dpi than in goats euthanized at 30 dpi, while the T. gondii concentration in hearts decreased. This study describes for the first time distribution of T. gondii parasites in post-weaned goat kids. New information about T. gondii predilection sites in goats and about the progression of infection between 30 and 90 dpi was achieved.

• MeSH
• játra parazitologie   MeSH
• kosterní svaly parazitologie   MeSH
• kozy MeSH
• kvantitativní polymerázová řetězová reakce metody   veterinární   MeSH
• magnetismus MeSH
• mozek parazitologie   MeSH
• nemoci koz diagnóza   parazitologie   MeSH
• plíce parazitologie   MeSH
• slezina parazitologie   MeSH
• srdce parazitologie   MeSH
• Toxoplasma izolace a purifikace   MeSH
• toxoplazmóza zvířat diagnóza   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH