Several divergent Cryptosporidium parvum subtypes have emerged in people in recent years, but their infectivity, pathogenicity, and genetic characteristics are unclear. In the present study, IFN-γ knockout C57BL/6 (GKO) mice were infected with the novel IIoA15G1 and IIpA11 subtypes of C. parvum and the common IIaA17G2R1 subtype. The genomes of these isolates were sequenced and compared with each other. Further gene tagging and deletion were performed on the most polymorphic virulence-associated cgd8_5420 gene encoding a hypothetical protein using the CRISPR/Cas9 technology. IIpA11 and IIoA15G1 were highly infectious in GKO mice, with an ID50 of 2.4 and 3.6 oocysts, respectively. The duration of oocyst shedding for IIpA11 (>58.0 ± 1.4 d) and IIoA15G1 (>57.5 ± 0.9 d) was significantly longer than for IIaA17G2R1 (5.5 ± 0.9 d; p < 0.001). One of the mice infected with IIpA11 died on day 33 post infection. The genomes of IIaA17G2R1, IIoA15G1, and IIpA11 had 203, 46839, and 47,122 single nucleotide polymorphisms, respectively, compared to C. parvum IOWA II. In contrast, only 3,361 nucleotide differences were found between IIoA15G1 and IIpA11, with several genes encoding invasion-associated mucin glycoproteins and cgd8_5420 encoding a secretory protein being highly polymorphic. The latter is mainly expressed in trophozoites, merozoites, and macrogametes. Deletion of this gene reduced the intensity of IIpA11 infection and increased the survival of infected mice. Therefore, the emerging IIoA15G1 and IIpA11 subtypes have divergent genomes compared to common IIa subtypes and are highly infectious and pathogenic in GKO mice. Several secretory proteins, including a variant protein encoded by the subtelomeric cgd8_5420 gene, are associated with differences in virulence between the two subtypes.

• Klíčová slova
• CRISPR/Cas9, Cryptosporidium, animal model, genomics, virulence,
• MeSH
• Cryptosporidium parvum * genetika   patogenita   klasifikace   MeSH
• faktory virulence * genetika   metabolismus   MeSH
• genom protozoální MeSH
• kryptosporidióza * parazitologie   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• oocysty MeSH
• protozoální proteiny * genetika   metabolismus   MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• faktory virulence * MeSH
• protozoální proteiny * MeSH

Cryptosporidium mortiferum, a parasite transmitted by squirrels, is beginning to spread in Europe. C. mortiferum was detected in a symptomatic human. A 44-year-old man from the Czech Republic suffered from gastroenteritis characterized by abdominal pain, nausea, and loose stools. Molecular analyses confirmed the XIVaA20G2T1 subtype in the patient's stool. At the same time, the same subtype of C. mortiferum was detected in three red squirrels and two ground squirrels in the area where the patient lived. The intensity of the infection was significantly higher in the red squirrels that died, while the ground squirrels showed no symptoms. The results of the study indicate that red squirrels and ground squirrels are the reservoirs for the infection.

• Klíčová slova
• Cryptosporidium mortiferum, human, infection, squirrels,
• MeSH
• Cryptosporidium * genetika   izolace a purifikace   klasifikace   MeSH
• dospělí MeSH
• feces parazitologie   MeSH
• fylogeneze MeSH
• gastroenteritida parazitologie   MeSH
• kryptosporidióza * parazitologie   přenos   epidemiologie   diagnóza   MeSH
• lidé MeSH
• Sciuridae * parazitologie   MeSH
• zdroje nemoci parazitologie   MeSH
• zoonózy * parazitologie   přenos   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• Geografické názvy
• Česká republika MeSH
• Evropa MeSH

Our team's discovery of the link between chronic "latent" infection with Toxoplasma gondii (Nicolle et Manceaux, 1908) and suicidal behaviour, and our subsequent cross-diagnostic confirmatory work and mechanistic extensions, evolved from our neuroimmunology studies on affective and behavioural dysregulation exacerbated by allergic sensitisation and allergen exposure. Another root was studying behavioural changes and cytokine gene expression in the brain of rodents sensitised and exposed to aeroallergens. We "piggy-backed" our project funded to study coupling between aeroallergen sensitisation and exposure in patients with recurrent mood disorders, by measuring Toxoplasma gondii (T. gondii) antibodies in existing samples, and found associations between IgG serointensity and past suicide attempts. Successively, we then reported significant associations between T. gondii seropositivity and/or serointensity and suicidal behaviour in patients with schizophrenia in Germany, recent attempters in Sweden, and longitudinally in a cohort of Danish mothers. In the Danish mothers the exposure to T. gondii preceded self-directed harm and violent suicide attempts; the association was stronger with higher serointensity strata demonstrating a dose-effect. Furthermore, we identified links between T. gondii IgG and suicide endophenotypes of aggression and impulsivity in both individuals with no history of mental illness, and in patients with Intermittent Explosive Disorder (IED). We also found associations between T. gondii and risk factors of suicidal behaviour such as hopelessness and anhedonia in the Amish, depressive symptoms in pregnant women and women Veterans, frailty in older adults, and cognitive deficits in patients with bipolar disorder. Recently, we reported positive associations between T. gondii IgG serointensity with suicidal ideation, impulsivity, depression scores, and daytime dysfunction due to sleep problems in US Veterans who previously attempted suicide. Toxoplasma gondii emerged rather unexpectedly and then took over a considerable proportion of our neuroimmune research portfolio. It satisfied both intellectual appetites, and brought celebrations of discovery, with three systematic reviews and meta-analyses published to date, and a substantial majority of primary articles confirming our initial observations. Toxoplasma gondii also brought considerable frustrations, such as initial grant application setbacks, inability to completely demonstrate causality and, so far, prophylactic and therapeutic impotence for mental health applications in general. While we do not have, as of today, effective and safe treatments for chronic toxoplasmosis with demonstrated mental health benefits in immunocompetent hosts, there are reasons to be optimistic regarding future discoveries. These may include vaccines, novel medications using in silico exploration with biological confirmation, trials of reactivation prevention, as well as identification and targeting of mediating mechanisms. Yet the most justified reasons for optimism are the potential to apply machine learning (ML) and artificial intelligence (AI) methodologies to big data with a focus on interaction and causal inference. These novel approaches, utilising ML-weighted models that emulate randomised trials in electronic medical records, have the potential to reveal not only if T. gondii elevates risk and to what extent, but also for whom specifically, under which demographic, clinical and physiological circumstances, and what factors, or combinations thereof, might mitigate this risk.

• Klíčová slova
• aggression, depression, frailty, impulsivity, sleep impairment, suicidal self-directed violence, suicidality, suicide risk factors,
• MeSH
• lidé MeSH
• sebevražda * psychologie   MeSH
• Toxoplasma * fyziologie   MeSH
• toxoplazmóza * psychologie   diagnóza   komplikace   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• úvodní články MeSH

Culicoides (Diptera: Ceratopogonidae) biting midges are a diverse group of insect vectors that transmit pathogens affecting humans, livestock, and wild animals. Among them, Oropouche virus, African Horse sickness virus, and bluetongue virus are the most notable pathogens. However, comparatively little is known about which Culicoides species serve as vectors of wildlife parasites affecting wild birds globally, including the malaria-like parasite of the genus Haemoproteus (Haemosporida: Haemoproteidae) and kinetoplastid Trypanosoma (Trypanosomatida: Trypanosomatidae). Beyond the direct impact of their bites, infections by these parasites negatively affect wild birds from early developmental stages, significantly influencing their ecology and evolution. Here, we present a comprehensive review of the role of Culicoides species in the transmission of these two genera of avian parasites in Europe: Haemoproteus and Trypanosoma. We identify key information and methods used to study Culicoides-bird-parasite interactions, from insect sampling to vector competence assessment. Additionally, we highlight key knowledge gaps and propose future research directions in this area.

• Klíčová slova
• Culicoides, Haemoproteus, Trypanosoma, Avian malaria, Birds, Blood parasites, Vectors,
• MeSH
• Ceratopogonidae * parazitologie   fyziologie   MeSH
• divoká zvířata parazitologie   MeSH
• Haemosporida fyziologie   izolace a purifikace   MeSH
• hmyz - vektory * parazitologie   MeSH
• nemoci ptáků * přenos   parazitologie   epidemiologie   MeSH
• protozoální infekce zvířat * přenos   parazitologie   epidemiologie   MeSH
• ptáci * parazitologie   MeSH
• Trypanosoma fyziologie   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Geografické názvy
• Evropa epidemiologie   MeSH

A metabolic disease resulting in elevated blood glucose levels, type-2 diabetes affects approximately 462 million people globally. Although its prevalence appears to be increasing, type-2 diabetes has been associated with various potentially preventable risk factors, including infectious diseases. The protozoal infection with Toxoplasma gondii (Nicolle et Manceaux, 1908) has been associated with type-2 diabetes in two previous meta-analyses. Since the publication of the last meta-analysis supporting an association between type-2 diabetes and T. gondii, several new primary studies have investigated this association. In this meta-analysis, we sought to further characterise the association between type-2 diabetes and T. gondii. We identified primary studies using PubMed, Embase, Scopus and Web of Science. Twenty-five studies met our inclusion criteria for a total of 4,639 patients with type-2 diabetes and 3,492 controls. Eighteen primary studies found a positive association between type-2 diabetes and T. gondii, whereas seven did not. Using a frequentist random-effects meta-analysis model, we found an overall summary odds ratio of 2.77 (95-percent confidence interval: 2.03-3.76), suggesting that the odds people will have type-2 diabetes is 2.7 times higher for people seropositive for T. gondii. Future studies should investigate this association in additional geographical regions and explore whether this association is due to the immunosuppressive effects of type-2 diabetes or whether T. gondii directly or indirectly affects glucose metabolism, or both.

• Klíčová slova
• diabetes mellitus, insulin-resistant diabetes, metabolic disorder, protozoal infection, toxoplasmosis,
• MeSH
• diabetes mellitus 2. typu * komplikace   parazitologie   MeSH
• lidé MeSH
• rizikové faktory MeSH
• Toxoplasma * imunologie   MeSH
• toxoplazmóza * komplikace   epidemiologie   parazitologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• metaanalýza MeSH
• systematický přehled MeSH

BACKGROUND: Ticks in Europe comprise over 100 species, with Ixodes, Rhipicephalus, Dermacentor, and Haemaphysalis being the most prevalent. Ixodes ricinus is the most widespread, while Dermacentor reticulatus is common in central Europe. Rhipicephalus sanguineus has expanded into temperate regions, affecting tick-host interactions and pathogen transmission. Companion animals, especially dogs and cats, are common tick hosts and vectors for pathogens such as borreliosis, anaplasmosis, and canine babesiosis. This study investigates the prevalence and species composition of ticks infesting dogs and cats in Poland, the Czech Republic, Slovakia, Hungary, and Romania, focusing on seasonal trends and pathogen occurrence. METHODS: From 2020 to 2022, ticks were collected from dogs and cats during veterinary consultations across five countries. A total of 4563 ticks were identified morphologically, and 1085 were screened for pathogens, including Babesia canis, Anaplasma phagocytophilum, Ehrlichia canis, and Borrelia burgdorferi sensu lato. RESULTS: I. ricinus was the predominant species (65.0%), followed by D. reticulatus (29.8%) and R. sanguineus (3.8%). Ticks were present year-round, with peaks in spring and autumn. Of the 1085 tested ticks, 29.0% were positive for at least one pathogen, with A. phagocytophilum being the most common (15.0%). B. burgdorferi s.l. was detected in 7.0% of ticks, and Babesia spp. in 6.5%, predominantly B. canis. CONCLUSIONS: This study highlights the year-round risk of tick infestations and vector-borne pathogen transmission in dogs and cats in Central Europe, underscoring the need for ongoing tick surveillance. Veterinary practitioners should enhance public awareness about ticks and associated health risks for companion animals.

• Klíčová slova
• Dermacentor reticulatus, Ixodes hexagonus, Ixodes ricinus, Cat, Central European Region, Companion animals, Dog, Ticks, Vector-borne,
• MeSH
• Anaplasma phagocytophilum izolace a purifikace   MeSH
• Babesia izolace a purifikace   MeSH
• biodiverzita MeSH
• Borrelia burgdorferi izolace a purifikace   MeSH
• infestace klíšťaty * veterinární   epidemiologie   parazitologie   MeSH
• Ixodidae * mikrobiologie   klasifikace   MeSH
• klíšťata * mikrobiologie   klasifikace   parazitologie   MeSH
• kočky parazitologie   MeSH
• nemoci koček * parazitologie   epidemiologie   MeSH
• nemoci přenášené klíšťaty * veterinární   epidemiologie   mikrobiologie   MeSH
• nemoci psů * parazitologie   epidemiologie   MeSH
• psi parazitologie   MeSH
• roční období MeSH
• zvířata MeSH
• Check Tag
• kočky parazitologie   MeSH
• mužské pohlaví MeSH
• psi parazitologie   MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Maďarsko epidemiologie   MeSH
• Polsko epidemiologie   MeSH
• Rumunsko epidemiologie   MeSH
• Slovenská republika epidemiologie   MeSH

BACKGROUND: Traditionally, blood meal analysis has been the primary method used to assess feeding patterns of insects. In contrast, parasite detection is commonly applied to monitor parasite circulation and prevalence in vectors, but rarely to study host feeding patterns. Our study aimed to test whether broad-target screening for haemosporidian and trypanosome parasites could complement blood barcoding by revealing additional host associations. We hypothesised that combining both methods would provide a more comprehensive understanding of vector feeding behaviour than either method alone. In addition to evaluating the two methods, we also analysed the vector species composition and their abundance, providing important faunistic and prevalence data that contribute to the broader understanding of local vector-parasite dynamics. METHODS: Mosquitoes and biting midges were trapped over a 5-year period at three localities in Czechia. Blood-fed individuals underwent blood meal barcoding analysis. In parallel, parasite detection was conducted using nested polymerase chain reaction (PCR) and gut dissection techniques. RESULTS: A total of 10,152 mosquitoes were collected, with Culex pipiens (66%) and Aedes vexans (18%) being the predominant species. In addition, 1701 biting midges, primarily Culicoides pictipennis (61%) and C. festivipennis (12%), were captured. Among the collected samples, 281 mosquitoes (3%) and 52 biting midges (3%) were blood-fed. Parasites were detected in 468 mosquito pools (5%, 341 trypanosomes, 127 haemosporidians) and 21 midge pools (1%, 8 trypanosomes, 13 haemosporidians). Blood meal barcoding of engorged Aedes, Anopheles, Culiseta, and Mansonia samples revealed only mammalian hosts; however, parasite detection indicated previous feeding on birds. Culex displayed stronger ornithophily according to parasite detection, although blood meal analysis showed a more opportunistic behaviour, with the detection of avian, mammalian and even amphibian blood. Avian parasites were detected in five Culicoides species (Culicoides alazanicus, C. festivipennis, C. kibunensis, C. nubeculosus and C. pictipennis) while human blood was detected only in C. pictipennis. Overall, four Haemoproteus lineages and 15 Plasmodium lineages were identified, 11 of which were new records for Czechia and 4 were newly described. CONCLUSIONS: Integrating blood meal analysis with parasite detection provides a more comprehensive understanding of insect feeding patterns and vector-host dynamics. Blood meal analysis remains the gold standard for identifying recent host interactions, offering direct and often species-level evidence of feeding events. In addition, parasite detection extends the window of detectability beyond the digestion of host blood and can reveal additional or otherwise-overlooked host associations. Together, these complementary approaches increase the likelihood of detecting interactions with a broader range of hosts, including humans, who might be missed by parasite screening alone.

• Klíčová slova
• T. theileri, Avian trypanosomes, Biting midge, Blood meal, Haemosporidians, Host feeding patterns, Method comparison, Mosquito,
• MeSH
• Ceratopogonidae * parazitologie   fyziologie   MeSH
• Culicidae * parazitologie   fyziologie   MeSH
• Haemosporida izolace a purifikace   genetika   MeSH
• hmyz - vektory * parazitologie   fyziologie   MeSH
• interakce hostitele a parazita MeSH
• komáří přenašeči * parazitologie   fyziologie   MeSH
• krev * parazitologie   MeSH
• polymerázová řetězová reakce MeSH
• stravovací zvyklosti * MeSH
• Trypanosoma izolace a purifikace   genetika   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Toxoplasma gondii (Nicolle et Manceaux, 1908), an intracellular parasite that causes toxoplasmosis, infects a third of the human population. Latent toxoplasmosis has been linked to altered immune responses, including elevated proinflammatory cytokines. In early pregnancy, the immune system adapts to balance inflammation and foetal tolerance. This study assessed whether pregnant women in the first trimester infected with Toxoplasma gondii have different cytokine levels than uninfected women. This study also examined whether women with discordant test results for toxoplasmosis represent a distinct group or a mixed group composed of infected women with unusually low levels of anti-Toxoplasma antibodies and uninfected women with high levels of cross-reacting antibodies. We measured 18 cytokines (IL-1β, IL-1ra, IL-2, IL-4, IL-7, IL-9, IL-17A, Eotaxin, FGF basic, G-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF-BB, RANTES, TNF-α) in 78 pregnant women, classified as Toxoplasma-positive, Toxoplasma-negative or Toxoplasma-discordant (negative by IgG ELISA, positive by complement fixation test [CFT]). Using exploratory factor analysis, we identified two factors, the first explaining 29.6% and the second 24.9% of the total variability in cytokine concentrations. Toxoplasma-positive women scored significantly higher in the second factor, primarily associated with cytokines linked to Th1-driven inflammation and cellular immunity. Specifically, these women exhibited elevated levels of IL-1β, IL-1ra, IL-2, FGF basic and PDGF-BB compared to Toxoplasma-negative women. This finding suggests that pregnant women with latent toxoplasmosis experience some degree of chronic inflammation. Additionally, our results indicate that Toxoplasma-discordant women are likely Toxoplasma-negative individuals with detectable anti-Toxoplasma IgM antibodies. However, as this study focused on pregnant women, further research is necessary to validate these conclusions in broader populations.

• Klíčová slova
• Toxoplasma gondii, cytokines, immunity, pregnancy,
• MeSH
• cytokiny krev   MeSH
• dospělí MeSH
• latentní infekce * MeSH
• lidé MeSH
• mladý dospělý MeSH
• parazitární komplikace těhotenství * imunologie   diagnóza   MeSH
• protilátky protozoální krev   MeSH
• sérologické testy MeSH
• těhotenství MeSH
• Toxoplasma imunologie   MeSH
• toxoplazmóza * diagnóza   imunologie   MeSH
• zánět * imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cytokiny MeSH
• protilátky protozoální MeSH

Toxoplasma gondii, an apicomplexan parasite and agent of the human disease toxoplasmosis, possesses a non-photosynthetic relic plastid, named the apicoplast. Thought to be evolved from a red algal plastid, the apicoplast houses major metabolic pathways, such as heme, isoprenoid and lipid synthesis, crucial for parasite survival, and thus considered attractive drug targets. However, despite similarities with plant chloroplast lipid synthesis pathways, the apicoplast lacks canonical plant/chloroplast lipid transporters and so metabolite import/export is at present, poorly characterised. Here we identify TgFLP12, a newly identified P5-ATPase transporter localised to the Toxoplasma apicoplast. TgFLP12 is found in the SAR (Stramenopile-Alveolata-Rhizaria) supergroup (to which belong Apicomplexa parasites and chromerids) but absent in higher plants. Disruption of TgFLP12 causes major defects on apicoplast morphology. Lipidomic analyses and stable isotope labelling reveal a unique accumulation of C14:0 in the apicoplast, which is then lacking in most major lipid classes subsequently synthesized in the ER. Successful complementation of a yeast mutant deficient in fatty acid transport with TgFLP12 validates TgFLP12 as a fatty acid transporter. Overall, we identify a potentially important drug target: the apicoplast fatty acid exporter, specific to Apicomplexa which unexpectedly also highlights Toxoplasma's utility as a model organism for investigating algal biology.

• MeSH
• adenosintrifosfatasy * metabolismus   genetika   MeSH
• apikoplasty * metabolismus   MeSH
• biologický transport MeSH
• chloroplasty * metabolismus   MeSH
• fylogeneze MeSH
• mastné kyseliny * metabolismus   MeSH
• metabolismus lipidů MeSH
• plastidy metabolismus   MeSH
• protozoální proteiny * metabolismus   genetika   MeSH
• Toxoplasma * metabolismus   genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adenosintrifosfatasy * MeSH
• mastné kyseliny * MeSH
• protozoální proteiny * MeSH

BackgroundMost Toxoplasma gondii infections in humans are considered foodborne, but the relative importance of the various routes of infection is largely unknown. Consumption of green produce contaminated with T. gondii oocysts has been identified as a possible source.AimWe aimed to estimate the occurrence and prevalence of T. gondii oocysts in commercially available ready-to-eat (RTE) salad mixes in 10 European countries.MethodsA real-time PCR-based method for oocyst detection was developed and optimised by two laboratories and validated in an interlaboratory test. This detection method and a harmonised sampling strategy were applied in a multi-country study. Multivariable logistic regression was used to investigate risk factors for oocyst contamination of RTE salad.ResultsThe real-time PCR method had a detection limit of 10 oocysts per 30 g of salad. We collected 3,329 RTE salad samples (baby leaf and cut leaf mixes) from October 2021 to September 2022. The prevalence of T. gondii oocyst contamination was 4.1% (95% confidence interval (CI): 3.4-4.8%; n = 3,293). In multivariable regression analysis, winter season, sampling and packaging of salad in Northern Europe and production of salad in Western Europe were associated with detection of T. gondii, with no statistically significant differences between salad types.ConclusionWe estimated the prevalence of T. gondii oocysts in RTE leafy green salads using a validated and standardised procedure to assess the potential risk for human infection; highlighting the need to address this risk at each critical point of the salad production chain.

• Klíčová slova
• Europe, Leafy greens, Toxoplasma gondii, harmonisation, oocysts, ready-to-eat salads, ring-trial,
• MeSH
• kontaminace potravin * analýza   MeSH
• kvantitativní polymerázová řetězová reakce * metody   MeSH
• lidé MeSH
• oocysty izolace a purifikace   MeSH
• potravinářská parazitologie * metody   MeSH
• prevalence MeSH
• Toxoplasma * izolace a purifikace   genetika   MeSH
• toxoplazmóza * epidemiologie   MeSH
• zelenina * parazitologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa epidemiologie   MeSH