Growing incidence of bacteria resistant to last-line antibiotics represents one of the most important medical issue, complicating the successful treatment of life-threatening infections. In this project, molecular-epidemiological and genomic analysis of carbapenemase-producing Enterobacterales (CPE) isolated from patients in Czech hospitals and its comparison with the carbapenem-susceptible population will be performed. Whole genome sequencing and bioinformatics analysis will allow to identify specific features of CPE, high-risk clones with increased importance for public health and mobile genetic elements associated with the dissemination of resistance to carbapenems in hospitals. Complex genomic analysis will reveal genetic structure of the studied bacterial populations including their dynamics through microevolutionary changes in the clinical context of a hospital outbreak. The project outcomes will be used to improve the diagnostics of infectious diseases and to outline effective interventions for the prevention and control of the transmission of bacterial pathogens.

Narůstající incidence bakterií rezistentních k antibiotikům poslední volby představuje jeden z nejvýznamnějších medicínských problémů, který komplikuje úspěšnou léčbu život ohrožujících infekcí. V rámci navrhovaného projektu bude provedena molekulárně-epidemiologická/genomická analýza Enterobacterales produkujících karbapenemázy (CPE) izolovaných z pacientů hospitalizovaných v českých nemocnicích a srovnání s bakteriální populací citlivou ke karbapenemům. S využitím nejmodernějších postupů celogenomového sekvenování a bioinformatické analýzy budou identifikovány jedinečné rysy CPE, vysoce rizikové klony se zvýšeným významem pro zdraví populace a mobilní genetické elementy spojené s šířením rezistence ke karbapenemům v nemocnicích. Komplexní genomická analýza umožní popsat genetickou strukturu sledovaných bakteriálních populací včetně její dynamiky prostřednictvím mikroevolučních změn během šíření nemocničních nákaz. Výsledky projektu budou bezprostředně využity pro zlepšení a zpřesnění diagnostiky infekčních nemocí a návrh účinnějších postupů v prevenci a kontrole šíření patogenů.

• Klíčová slova
• Whole-genome sequencing, antibiotic resistance, antibiotická rezistence, karbapenemy, populační studie, carbapenems, genomika, genomics, Enterobacterales, celogenomová sekvenace, Enterobacterales, population study,

• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Ticks are vectors of pathogens affecting human and animal health worldwide. Nevertheless, the ecological and evolutionary interactions between ticks, hosts, and pathogens are largely unknown. Here, we integrated a framework to evaluate the associations of the tickIxodes ricinuswith its hosts and environmental niches that impact pathogen circulation. The analysis of tick-hosts association suggested that mammals and lizards were the ancestral hosts of this tick species, and that a leap to Aves occurred around 120 M years ago. The signature of the environmental variables over the host's phylogeny revealed the existence of two clades of vertebrates diverging along a temperature and vegetation split. This is a robust proof that the tick probably experienced a colonization of new niches by adapting to a large set of new hosts, Aves. Interestingly, the colonization of Aves as hosts did not increase significantly the ecological niche ofI. ricinus, but remarkably Aves are super-spreaders of pathogens. The disparate contribution of Aves to the tick-host-pathogen networks revealed thatI. ricinusevolved to maximize habitat overlap with some hosts that are super-spreaders of pathogens. These results supported the hypothesis that large host networks are not a requirement of tick survival but pathogen circulation. The biological cost of tick adaptation to non-optimal environmental conditions might be balanced by molecular mechanisms triggered by the pathogens that we have only begun to understand.

• MeSH
• biologická adaptace MeSH
• druhová specificita MeSH
• ekologie metody   MeSH
• interakce hostitele a patogenu MeSH
• ještěři parazitologie   MeSH
• klíšťata mikrobiologie   parazitologie   fyziologie   MeSH
• klíště klasifikace   parazitologie   fyziologie   MeSH
• lidé MeSH
• nemoci přenášené klíšťaty parazitologie   přenos   MeSH
• obratlovci klasifikace   parazitologie   MeSH
• ptáci klasifikace   parazitologie   MeSH
• savci klasifikace   parazitologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• Anaplasma phagocytophilum MeSH
• biologická adaptace genetika   fyziologie   MeSH
• biologická evoluce MeSH
• ekologie * MeSH
• genetická transkripce genetika   MeSH
• interakce hostitele a patogenu genetika   MeSH
• klíšťata mikrobiologie   MeSH
• klíště mikrobiologie   MeSH
• molekulární evoluce MeSH
• regulace genové exprese genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The life cycle of spirochetes of the genus Borrelia includes complex networks of vertebrates and ticks. The tripartite association of Borrelia-vertebrate-tick has proved ecologically successful for these bacteria, which have become some of the most prominent tick-borne pathogens in the northern hemisphere. To keep evolutionary pace with its double-host life history, Borrelia must adapt to the evolutionary pressures exerted by both sets of hosts. In this review, we attempt to reconcile functional, phylogenetic, and ecological perspectives to propose a coherent scenario of Borrelia evolution. Available empirical information supports that the association of Borrelia with ticks is very old. The major split between the tick families Argasidae-Ixodidae (dated some 230-290 Mya) resulted in most relapsing fever (Rf) species being restricted to Argasidae and few associated with Ixodidae. A further key event produced the diversification of the Lyme borreliosis (Lb) species: the radiation of ticks of the genus Ixodes from the primitive stock of Ixodidae (around 217 Mya). The ecological interactions of Borrelia demonstrate that Argasidae-transmitted Rf species remain restricted to small niches of one tick species and few vertebrates. The evolutionary pressures on this group are consequently low, and speciation processes seem to be driven by geographical isolation. In contrast to Rf, Lb species circulate in nested networks of dozens of tick species and hundreds of vertebrate species. This greater variety confers a remarkably variable pool of evolutionary pressures, resulting in large speciation of the Lb group, where different species adapt to circulate through different groups of vertebrates. Available data, based on ospA and multilocus sequence typing (including eight concatenated in-house genes) phylogenetic trees, suggest that ticks could constitute a secondary bottleneck that contributes to Lb specialization. Both sets of adaptive pressures contribute to the resilience of highly adaptable meta-populations of bacteria.

• MeSH
• biologická adaptace MeSH
• biologická evoluce * MeSH
• Borrelia klasifikace   fyziologie   MeSH
• infekce přenášené vektorem * MeSH
• interakce hostitele a patogenu MeSH
• klíšťata mikrobiologie   MeSH
• lidé MeSH
• lymeská nemoc mikrobiologie   přenos   MeSH
• selekce (genetika) MeSH
• zdroje nemoci * mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

BACKGROUND: Although many genetic manipulations of crops providing biofortified or safer food have been done, the acceptance of biotechnology crops still remains limited. We report on a transgenic barley expressing the multi-functional protein osmotin that improves plant defense under stress conditions. METHODS: An Agrobacterium-mediated technique was used to transform immature embryos of the spring barley cultivar Golden Promise. Transgenic barley plants of the T0 and T1 generation were evaluated by molecular methods. Transgenic barley tolerance to stress was determined by chlorophyll, total protein, malondialdehyde and ascorbate peroxidase content. Methanol extracts of i) Fusarium oxysporum infected or ii) salt-stressed plants, were characterized by their acute toxicity effect on human dermal fibroblasts (HDF), genotoxicity and affection of biodiversity interactions, which was tested through monitoring barley natural virus pathogen-host interactions-the BYDV and WDV viruses transmitted to the plants by aphids and leafhoppers. RESULTS: Transgenic plants maintained the same level of chlorophyll and protein, which significantly declined in wild-type barley under the same stressful conditions. Salt stress evoked higher ascorbate peroxidase level and correspondingly less malondialdehyde. Osmotin expressing barley extracts exhibited a lower cytotoxicity effect of statistical significance than that of wild-type plants under both types of stress tested on human dermal fibroblasts. Extract of Fusarium oxysporum infected transgenic barley was not able to damage DNA in the Comet assay, which is in opposite to control plants. Moreover, this particular barley did not affect the local biodiversity. CONCLUSION: Our findings provide a new perspective that could help to evaluate the safety of products from genetically modified crops.

• MeSH
• bezpečnost potravin * MeSH
• biologická adaptace MeSH
• ektopická exprese * MeSH
• fyziologický stres genetika   MeSH
• geneticky modifikované rostliny MeSH
• interakce hostitele a patogenu genetika   MeSH
• ječmen (rod) genetika   MeSH
• lidé MeSH
• rekombinantní proteiny * MeSH
• rostlinné proteiny genetika   MeSH
• tabák genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

To date, most molecular investigations of schistosomatids have focused principally on blood flukes (schistosomes) of humans. Despite the clinical importance of cercarial dermatitis in humans caused by Trichobilharzia regenti and the serious neuropathologic disease that this parasite causes in its permissive avian hosts and accidental mammalian hosts, almost nothing is known about the molecular aspects of how this fluke invades its hosts, migrates in host tissues and how it interacts with its hosts' immune system. Here, we explored selected aspects using a transcriptomic-bioinformatic approach. To do this, we sequenced, assembled and annotated the transcriptome representing two consecutive life stages (cercariae and schistosomula) of T. regenti involved in the first phases of infection of the avian host. We identified key biological and metabolic pathways specific to each of these two developmental stages and also undertook comparative analyses using data available for taxonomically related blood flukes of the genus Schistosoma. Detailed comparative analyses revealed the unique involvement of carbohydrate metabolism, translation and amino acid metabolism, and calcium in T. regenti cercariae during their invasion and in growth and development, as well as the roles of cell adhesion molecules, microaerobic metabolism (citrate cycle and oxidative phosphorylation), peptidases (cathepsins) and other histolytic and lysozomal proteins in schistosomula during their particular migration in neural tissues of the avian host. In conclusion, the present transcriptomic exploration provides new and significant insights into the molecular biology of T. regenti, which should underpin future genomic and proteomic investigations of T. regenti and, importantly, provides a useful starting point for a range of comparative studies of schistosomatids and other trematodes.

• MeSH
• biologická adaptace * MeSH
• interakce hostitele a patogenu * MeSH
• kachny parazitologie   MeSH
• metabolické sítě a dráhy genetika   MeSH
• molekulární sekvence - údaje MeSH
• Schistosomatidae genetika   růst a vývoj   MeSH
• sekvenční analýza DNA MeSH
• stadia vývoje MeSH
• stanovení celkové genové exprese * MeSH
• výpočetní biologie * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Gregarines represent a very diverse group of early emerging apicomplexans, parasitising numerous invertebrates and urochordates, and are considered of little practical significance. Recently, they have gained more attention since some analyses showed that cryptosporidia are more closely related to the gregarines than to coccidia. METHODOLOGY/PRINCIPAL FINDINGS: Using a combined microscopic approach, this study points out the spectacular strategy of Gregarina cuneata for attachment to host tissue and nutrient acquisition while parasitising the intestine of yellow mealworm larvae, and reveals the unusual dynamics of cellular interactions between the host epithelium and parasite feeding stages. Trophozoites of G. cuneata develop epicellularly, attached to the luminal side of the host epithelial cell by an epimerite exhibiting a high degree of morphological variability. The presence of contractile elements in the apical region of feeding stages indicates that trophozoite detachment from host tissue is an active process self-regulated by the parasite. A detailed discussion is provided on the possibility of reversible retraction and protraction of the eugregarine apical end, facilitating eventual reattachment to another host cell in better physiological conditions. The gamonts, found in contact with host tissue via a modified protomerite top, indicate further adaptation of parasite for nutrient acquisition via epicellular parasitism while keeping their host healthy. The presence of eugregarines in mealworm larvae even seems to increase the host growth rate and to reduce the death rate despite often heavy parasitisation. CONCLUSIONS/SIGNIFICANCE: Improved knowledge about the formation of host-parasite interactions in deep-branching apicomplexans, including gregarines, would offer significant insights into the fascinating biology and evolutionary strategy of Apicomplexa. Gregarines exhibit an enormous diversity in cell architecture and dimensions, depending on their parasitic strategy and the surrounding environment. They seem to be a perfect example of a coevolution between a group of parasites and their hosts.

• MeSH
• aktiny metabolismus   MeSH
• Apicomplexa cytologie   fyziologie   ultrastruktura   MeSH
• epitel metabolismus   parazitologie   ultrastruktura   MeSH
• epitelové buňky metabolismus   parazitologie   ultrastruktura   MeSH
• fyziologická adaptace * MeSH
• interakce hostitele a patogenu MeSH
• myosiny metabolismus   MeSH
• symbióza * MeSH
• Tenebrio parazitologie   MeSH
• transport proteinů MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Klíčová slova
• adaptace patogenu, multiplexová analýza,
• MeSH
• acelulární vakcíny aplikace a dávkování   imunologie   MeSH
• bakteriologické techniky MeSH
• Bordetella parapertussis izolace a purifikace   MeSH
• Bordetella pertussis * imunologie   izolace a purifikace   patogenita   MeSH
• dítě MeSH
• dospělí MeSH
• ELISA MeSH
• faktory virulence rodu Bordetella MeSH
• imunita získaná od matky imunologie   MeSH
• incidence MeSH
• infekce bakteriemi rodu Bordetella diagnóza   imunologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• novorozenec MeSH
• očkovací schéma MeSH
• odběr biologického vzorku MeSH
• pertuse * diagnóza   epidemiologie   imunologie   prevence a kontrola   MeSH
• pertusová vakcína aplikace a dávkování   imunologie   MeSH
• pertusový toxin imunologie   MeSH
• protilátky bakteriální imunologie   krev   MeSH
• sekundární imunizace MeSH
• sérologické testy metody   MeSH
• těhotenství imunologie   MeSH
• vakcína proti záškrtu, tetanu a černému kašli aplikace a dávkování   imunologie   MeSH
• vakcinace MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• novorozenec MeSH
• těhotenství imunologie   MeSH
• ženské pohlaví MeSH

Viroid-caused pathogenesis is a specific process dependent on viroid and host genotype(s), and may involve viroid-specific small RNAs (vsRNAs). We describe a new PSTVd variant C3, evolved through sequence adaptation to the host chamomile (Matricaria chamomilla) after biolistic inoculation with PSTVd-KF440-2, which causes extraordinary strong ('lethal') symptoms. The deletion of a single adenine A in the oligoA stretch of the pathogenicity (P) domain appears characteristic of PSTVd-C3. The pathogenicity and the vsRNA pool of PSTVd-C3 were compared to those of lethal variant PSTVd-AS1, from which PSTVd-C3 differs by five mutations located in the P domain. Both lethal viroid variants showed higher stability and lower variation in analyzed vsRNA pools than the mild PSTVd-QFA. PSTVd-C3 and -AS1 caused similar symptoms on chamomile, tomato, and Nicotiana benthamiana, and exhibited similar but species-specific distributions of selected vsRNAs as quantified using TaqMan probes. Both lethal PSTVd variants block biosynthesis of lignin in roots of cultured chamomile and tomato. Four 'expression markers' (TCP3, CIPK, VSF-1, and VPE) were selected from a tomato EST library to quantify their expression upon viroid infection; these markers were strongly downregulated in tomato leaf blades infected by PSTVd-C3- and -AS1 but not by PSTVd-QFA.

• MeSH
• fyziologická adaptace * MeSH
• genetické markery genetika   MeSH
• interakce hostitele a patogenu MeSH
• lignin metabolismus   MeSH
• malá nekódující RNA genetika   MeSH
• Matricaria virologie   MeSH
• messenger RNA genetika   metabolismus   MeSH
• molekulární evoluce * MeSH
• molekulární sekvence - údaje MeSH
• RNA virová genetika   MeSH
• sekvence nukleotidů MeSH
• Solanum lycopersicum virologie   MeSH
• Solanum tuberosum metabolismus   virologie   MeSH
• termodynamika MeSH
• viroidy genetika   patogenita   fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Cíl studie: Cílem bylo získat informace o klinickém průběhu matek pozitivních na virus SARS-CoV-2 a jejich novorozenců v období první vlny pandemie nemoci COVID-19 v České republice (1. 3.–31. 8. 2020). Design studie: Retrospektivní epidemiologická studie. Soubory: Soubor tvořilo 18 žen pozitivních na SARS-CoV-2 a stejný počet novorozenců. Sběr dat: Sběr dat probíhal formou otázek zaslaných e-mailem, případně doplněných telefonicky, na předem zvolená gynekologicko-porodnická a neonatologická pracoviště v České republice (n = 90). Výsledky: Z celkového počtu 18 žen s pozitivitou SARS-CoV-2 v nazofaryngeálním stěru bylo 15 rodiček asymptomatických, 2 ženy měly mírné příznaky a pouze 1 měla závažné klinické projevy. Všichni novorozenci se narodili v rozmezí od 36. do 41. gestačního týdne, ve 12 případech (67,7 %) se jednalo o spontánní vaginální porod, 6 novorozenců (32,3 %) se narodilo císařským řezem. Průměrná porodní hmotnost byla 3275 g, u žádného novorozence nebyla stanovena diagnóza intrauterinní růstové restrikce. Poporodní adaptace probíhala bez komplikací, pouze v jednom případě se rozvinul lehký RDS (syndrom dechové tísně). U 13 novorozenců byl proveden výtěr z nazofaryngu. Pozitivní nález byl pouze v 1 případě, a to u asymptomatického novorozence 14. den života. K rozvoji RDS došlo pouze u 1 novorozence v situaci, kdy nebylo možné vyloučit ev. podíl aspirace plodové vody, ostatní novorozenci (94,4 %) byli klinicky zcela v pořádku. Závěr: Shromážděná data z České republiky v průběhu první vlny pandemie potvrzují závěry současných studií ukazujících, že infekce SARS-CoV-2 u těhotných žen ve většině případů nezpůsobuje nepříznivé následky u novorozenců. Z literatury vyplývá, že naprostá většina matek prodělá mírné až středně závažné příznaky a prognóza novorozenců je dobrá, což je v souladu s výsledky naší studie. Současně u sledovaných žen nedošlo k vertikálnímu přenosu infekce.

Object of the study: Object of the study was to obtain information about clinical state of COVID-19 positive mothers and their newborns during the first wave of COVID-19 pandemic in the Czech Republic (1.3.–31.8.2020). Design of the study: Retrospective epidemiological study. Groups: Group of patients consisted of 18 SARS-CoV-2 positive pregnant women and the same amount of newborns. Data collection: Data was collected from previously designated gynaecology and obstetrics and neonatology departments in the Czech Republic (n=90) through set of questions sent via e-mail and when necessary completed through phone call. Outcomes: Out of 18 women with SARS-CoV-2 positive nasopharyngeal swab, 15 were asymptomatic, 2 women had mild symptoms and only 1 had severe clinical symptoms. All newborns were born between the 36th and 41st gestational week, 12 of them (67.7%) were born by spontaneous vaginal birth, 6 newborns (32.3%) were born via cesarean section. Average birth weight was 3275 g, no newborn was diagnosed with intrauterine growth restriction. Postnatal adaptation was uncomplicated except for one case of mild RDS (respiratory distress syndrome). 13 newborns underwent nasopharyngeal swab. Only 1 was positive in an asymptomatic newborn on the 14th day of life. RDS developed only in 1 newborn, there it was not able to rule out amniotic fluid aspiration, other newborns (94.4%) were clinically with no symptoms. Conclusion: Collected data from the Czech Republic during the first wave of pandemic confirms conclusions of up-to-date studies. SARS-CoV-2 infection in pregnant women has in majority of cases no adverse effect on their newborns. Majority of mothers overcame mild or moderate symptoms and prognosis of their newborns is good as reported in literature and also in unity with conclusion of our study. At the same time no vertical transmission of infection was reported.

• MeSH
• COVID-19 * MeSH
• infekční komplikace v těhotenství * MeSH
• lidé MeSH
• novorozenec MeSH
• retrospektivní studie MeSH
• SARS-CoV-2 MeSH
• těhotenství MeSH
• vertikální přenos infekce * MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• těhotenství MeSH
• ženské pohlaví MeSH