Silymarin is an extract obtained from the seeds of milk thistle (Sylibum marianum L., Asteraceae) and contains several structurally related flavonolignans and a small family of flavonoids. Mouse spleen cells represent highly sensitive primary cells suitable for studying the pharmacological potential and biofunctional properties of natural substances. Cultivation of splenocytes for 24 h under standard culture conditions (humidity, 37 °C, 5% CO2, atmospheric oxygen) resulted in decreased viability of splenocytes compared to intact cells. A cytoprotective effect of silybin (SB), silychristin (SCH) and 2,3-dehydrosilybin (DHSB) was observed at concentrations as low as 5 µmol/ml. At 50 µmol/ml, these substances restored and/or stimulated viability and mitochondrial membrane potential and had anti-apoptotic effect in the order SB > DHSB > SCH. The substances demonstrated a concentration-dependent activity in restoring the redox balance based on the changes in the concentration of reactive oxygen species (ROS), hydrogen peroxide (H2O2) and nitric oxide. This was in the order DHSB > SCH > SB, which correlated with the suppressed expression of nuclear factor erythroid 2-related factor 2 (Nrf2), catalase and glutathione peroxidase. The strong stimulation of the superoxide dismutase 1 gene converting ROS to H2O2 points to its dominant role in the maintaining redox homeostasis in splenocytes, which was disrupted by oxidative stress due to non-physiological culture conditions. Our study showed significant differences in the cytoprotective, antioxidant and anti-apoptotic activities of SB, SCH, and DHSB on splenocytes exposed to mild and AAPH-induced oxidative stress.
- Klíčová slova
- 2,3-dehydrosilybin, Apoptosis, Mouse splenocytes, Redox balance, Silybin, Silychristin, Viability,
- MeSH
- antioxidancia * farmakologie MeSH
- apoptóza * účinky léků MeSH
- cytoprotekce účinky léků MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- myši inbrední BALB C * MeSH
- myši MeSH
- oxidační stres účinky léků MeSH
- peroxid vodíku MeSH
- reaktivní formy kyslíku metabolismus MeSH
- silibinin * farmakologie MeSH
- silymarin * farmakologie MeSH
- slezina * cytologie účinky léků metabolismus MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antioxidancia * MeSH
- peroxid vodíku MeSH
- reaktivní formy kyslíku MeSH
- silibinin * MeSH
- silychristin MeSH Prohlížeč
- silymarin * MeSH
The diphyllobothriid tapeworm Dibothriocephalus dendriticus, one of the causative agents of the fish-borne zoonosis dibothriocephalosis, is mainly distributed in the Arctic/subarctic and temperate zones of the Northern Hemisphere (Europe, North America, and Asia), but also in the southern cone region of South America (Patagonia). The genetic structure and gene flow among 589 individuals of D. dendriticus, representing 20 populations, were studied using the mitochondrial cox1 gene as the first choice marker and 10 polymorphic nuclear microsatellite loci as a dominant molecular tool. The haplotype network of 30 cox1 haplotypes and Principal Coordinate Analysis/Structure analysis based on microsatellite data revealed close genetic relationships among populations within continents, namely northern and northwestern Europe (Norway, Finland, and UK/Scotland), North America (USA/Alaska, USA/Oregon, and Greenland), and South America (Argentina and Chile). The population from Iceland, located on the Mid-Atlantic Ridge between North America and Europe, was related to both Europe and North America. The mixed Nearctic and Palaearctic origin of D. dendriticus in Iceland was confirmed by a coalescent-based analysis of the microsatellite loci (in Migrate software). The most likely scenarios suggested that the Icelandic population is a genetic admixture of tapeworms from northwestern Europe and USA/Alaska. These findings corresponded with the distribution and migratory routes of piscivorous birds of the family Laridae, the main definitive hosts of D. dendriticus. The origin of an apparently non-native population of D. dendriticus in Patagonia was investigated in detail. The two most plausible hypotheses based on Migrate analysis represented different scenarios. One of them points to a European origin of the Patagonian population, the other to a North American origin. Future research with more extensive and a geographically broader sampling set is recommended to trace dispersal routes of D. dendriticus in Patagonia.
- Klíčová slova
- Broad tapeworm, Cox1, Diphyllobothriosis, Diphyllobothrium dendriticum, Fish-borne zoonosis, Microsatellite loci, Migratory routes, Mitochondrial DNA,
- Publikační typ
- časopisecké články MeSH
Lyme borreliosis (LB) is the most prevalent tick-borne human infection in Europe, with increasing incidence during the latest decades. Abundant populations of Ixodes ricinus, the main vector of the causative agent, spirochetes from the Borrelia burgdorferi sensu lato (Bbsl) complex, have been observed in urban and suburban areas of Europe, in general, and Slovakia, particularly. Understanding the spread of infectious diseases is crucial for implementing effective control measures. Global changes affect contact rates of humans and animals with Borrelia-infected ticks and increase the risk of contracting LB. The aim of this study was to investigate spatial and temporal variation in prevalence of Bbsl and diversity of its species in questing I. ricinus from three sites representing urban/suburban, natural and agricultural habitat types in Slovakia. Ixodes ricinus nymphs and adults were collected by dragging the vegetation in green areas of Bratislava town (urban/suburban habitat), in the Small Carpathians Mountains (natural habitat) (south-western Slovakia) and in an agricultural habitat at Rozhanovce in eastern Slovakia. Borrelia presence in ticks was detected by PCR and Bbsl species were identified by restriction fragment length polymorphism (RFLP). Borrelia burgdorferi s.l. species in coinfected ticks were identified by reverse line blot. Significant spatial and temporal variability in prevalence of infected ticks was revealed in the explored habitats. The lowest total prevalence was detected in the urban/suburban habitat, whereas higher prevalence was found in the natural and agricultural habitat. Six Bbsl species were detected by RFLP in each habitat type -B. burgdorferi sensu stricto (s.s.), B. afzelii, B. garinii, B. valaisiana, B. lusitaniae and B. spielmanii. Coinfections accounted for 3% of the total infections, whereby B. kurtenbachii was identified by RLB and sequencing in mixed infection with B. burgdorferi s.s, B. garinii and B. valaisiana. This finding represents the first record of B. kurtenbachii in questing I. ricinus in Slovakia and Europe. Variations in the proportion of Bbsl species were found between nymphs and adults, between years and between habitat types. Spatial variations in prevalence patterns and proportion of Bbsl species were also confirmed between locations within a relatively short distance in the urban habitat. Habitat-related and spatial variations in Borrelia prevalence and distribution of Bbsl species are probably associated with the local environmental conditions and vertebrate host spectrum. Due to the presence of Borrelia species pathogenic to humans, all explored sites can be ranked as areas with high epidemiological risk.
- Klíčová slova
- Borrelia burgdorferi, Ixodes ricinus, Slovakia, prevalence, species,
- Publikační typ
- časopisecké články MeSH
Dibothriocephalus latus is the most frequent causative agent of fish-borne zoonosis (diphyllobothriosis) in Europe, where it is currently circulating mainly in the Alpine lakes region (ALR) and Russia. Three mitochondrial genes (cox1, cob and nad3) and 6 microsatellite loci were analysed to determine how is the recently detected triploidy/parthenogenesis in tapeworms from ALR displayed at the DNA level. A geographically distant population from the Krasnoyarsk Reservoir in Russia (RU-KR) was analysed as a comparative population. One or 2 alleles of each microsatellite locus was detected in plerocercoids from RU-KR, corresponding to the microsatellite pattern of a diploid organism. In contrast, 1–3 alleles were observed in tapeworms from ALR, in accordance with their triploidy. The high diversity of mitochondrial haplotypes in D. latus from RU-KR implied an original and relatively stable population, but the identical structure of mitochondrial genes of tapeworms from ALR was probably a consequence of a bottleneck typical of introduced populations. These results indicated that the diploid/sexually reproducing population from RU-KR was ancestral, located within the centre of the distribution of the species, and the triploid/parthenogenetically reproducing subalpine population was at the margin of the distribution. The current study revealed the allelic structure of the microsatellite loci in the triploid tapeworm for the first time.
- Klíčová slova
- Asexual population, Diphyllobothriosis, fish-borne zoonosis, haplotypes, microsatellites, mitochondrial DNA, parthenogenesis, triploid tapeworms,
- MeSH
- Cestoda * genetika MeSH
- cestodózy * MeSH
- Diphyllobothrium * genetika MeSH
- genetická variace MeSH
- jezera MeSH
- lidé MeSH
- mikrosatelitní repetice MeSH
- triploidie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tapeworms of the genus Paracaryophyllaeus Kulakovskaya, 1961 (Cestoda: Caryophyllidea) are specific parasites of loaches (Cypriniformes: Cobitoidea) and occur almost exclusively in the Palaearctic region. The only exception and example of vicariance over the borders of two zoogeographical regions is Paracaryophyllaeus lepidocephali (Kundu, 1985), an insufficiently known species described from the Indomalayan region, with uncertain generic allocation. In the present paper, the species is redescribed based on new material collected from the type host, Lepidocephalichthys guntea, in West Bengal, India. Molecular data reveal this species as a member of Paracaryophyllaeus, within which it is the most closely related to P. cf. gotoi from Misgurnus anguillicaudatus in China and Japan on the basis of large subunit of ribosomal nuclear DNA (28S rDNA). Generic position of P. lepidocephali examined herein is also confirmed by morphology including cross sections, in particular, by a small, cylindrical body, medullary testes with testicular fields crossing the anterior margin of the cirrus sac, medullary vitelline follicles, with some follicles paramuscular, a shallow common genital atrium, short vagina and uterus not extending anteriorly beyond the cirrus sac. It differs from all Palaearctic congeners but Paracaryophyllaeus vladkae Scholz, Oros and Aydogdu, 2014 by the testicular field crossing the anterior margin of the cirrus sac. It differs from P. vladkae by more anterior position of the first vitelline follicles compared to the first testes. This species is a unique example of a fish tapeworm crossing the borders of the Palaearctic and Indomalayan zoogeographical regions.
- Klíčová slova
- Freshwater fishes, Molecular phylogeny, Tapeworms, Taxonomy, Zoogeography,
- MeSH
- Cestoda anatomie a histologie klasifikace genetika ultrastruktura MeSH
- cestodózy epidemiologie parazitologie veterinární MeSH
- fylogeneze MeSH
- máloostní parazitologie MeSH
- mikroskopie elektronová rastrovací veterinární MeSH
- nemoci ryb epidemiologie parazitologie MeSH
- RNA helmintů analýza MeSH
- RNA ribozomální 28S analýza MeSH
- rozšíření zvířat MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Indie epidemiologie MeSH
- Názvy látek
- RNA helmintů MeSH
- RNA ribozomální 28S MeSH
This work introduces new morphological and molecular information on the filaroid nematode Setaria cervi (Rudolphi, 1819) obtained from 13 infected game ungulates out of 96 dissected. The hosts comprised the following: a single moose (Alces alces), ten red deer (Cervus elaphus) and two sika deer (Cervus nippon) originating from the western and northern regions of the Czech Republic. Based on the complete sequences of the gene encoding mitochondrial cytochrome c oxidase subunit 1 (cox1), all 20 females and four males belonged to the species S. cervi. We detected three developmental female stages (adult fertile females, juvenile L5 females and L4 female larvae) differing in size and some morphological traits as the subtle structure of peribuccal crown and shape and features of tail knob. Such differences were described in detail for the first time. The phylogenetic relationships within the family Onchocercidae have been evaluated using new information on the cox1 sequence of S. cervi (maximum likelihood method, GTR + I + G model). In accordance with the latest phylogenetic studies, the present analysis confirmed the ancient separation of the subclass Setariinae from the remaining two onchocercid lineages Dirofilariinae and Onchocerinae.
- Klíčová slova
- Game ruminants, Setaria cervi larval stages, morphology, nematode, phylogenetic analysis, setariinae,
- MeSH
- larva anatomie a histologie genetika růst a vývoj MeSH
- proteiny červů analýza MeSH
- Setaria (Nematoda) anatomie a histologie enzymologie genetika růst a vývoj MeSH
- setariáza diagnóza parazitologie MeSH
- vysoká zvěř * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- proteiny červů MeSH
In Slovakia, the Roma population forms the second-largest ethnic minority. A large part of the Roma is semi-nomadic or lives in segregated settlements with poor living standards and limited access to health facilities. More than 40 years ago, a cross-sectional survey revealed a high prevalence of parasitic infections. There is a paucity of recent data, and hence, we designed a study to investigate the current status of intestinal parasitic infections in this population. Overall, 259 children aged 7 months to 18 years from 32 different segregated settlements provided faecal samples for microscopic examination using a sodium acetate-acetic acid-formalin concentration and the Paraprep L technique. Almost 40% of the samples yielded a positive result, with Ascaris lumbricoides (27.4%) and Giardia intestinalis (9.3%) being the most frequent helminth and intestinal protozoa species, respectively. Many children younger than 2 years were found to be infected, which suggests that community transmission is important. In view of our findings, there is a pressing need for targeted action to improve the health status of this neglected population.
- Klíčová slova
- Cross-sectional survey, Diagnosis, Intestinal parasites, Paraprep L, Roma, Slovakia, Sodium acetate-acetic acid-formalin (SAF) concentration technique,
- MeSH
- dítě MeSH
- etnicita MeSH
- feces parazitologie MeSH
- formaldehyd MeSH
- giardiáza epidemiologie parazitologie MeSH
- helmintóza epidemiologie parazitologie MeSH
- kojenec MeSH
- lidé MeSH
- menšiny MeSH
- mladiství MeSH
- paraziti MeSH
- předškolní dítě MeSH
- prevalence MeSH
- průřezové studie MeSH
- rizikové faktory MeSH
- zvířata MeSH
- Check Tag
- dítě MeSH
- kojenec MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Slovenská republika epidemiologie MeSH
- Názvy látek
- formaldehyd MeSH
BACKGROUND: A cytogenetic analysis of the new local triploid population of the caryophyllidean tapeworm Atractolytocestus huronensis, a unique parthenogenetic species with the ability to colonise new regions, was performed to understand the inner structure of its chromosome complement. METHODS: A karyotype analysis was carried out using classical Giemsa staining and C-banding combined with fluorescent DAPI staining. A hypothesis that triplets are composed from three homologue chromosomes of approximately the same length and same centromere position was tested statistically for multiple dependent variables using a non-parametric Friedman's ANOVA. The chromosomal location of ribosomal DNA clusters within the nucleolar organization region (NORs) and telomeric (TTAGGG)n sequences were detected by fluorescent in situ hybridization (FISH). Chromosomes were subjected to AgNO3 staining in order to determine whether the rDNA sites represent active NORs. RESULTS: The cytogenetic analysis confirmed the karyotype composed from eight chromosome triplets (3n = 24) as well as the existence of a pair of NORs located on each chromosome of the second triplet. Six NORs varied their activity from cell to cell, and it was reflected in the numbers of nucleoli (from 1 to 5). A huge morphological diversification of homologue chromosomes was originally detected in six out of eight triplets; the homologue elements differed significantly either in length and/or morphology, and some of them carried discernible interstitial telomeric sequences (ITSs), while the end telomeres were minute. The heterochromatin bands with high AT content varied irregularly, and the course of aberrant spermatogenesis was evident. CONCLUSIONS: Diversification of homologues is a unique phenomenon very likely caused by the long-term absence of a recombination and consequential accumulation of chromosome rearrangements in the genome of A. huronensis during species evolution. Unalterable asexual reproduction of the tapeworm, along with international trade in its host (carp), is facilitating its ongoing spread.
- Klíčová slova
- Aberrant meiosis, Cestoda, Diversification of homologues, Interstitial telomere sequences (ITSs), Polyploidy,
- MeSH
- Cestoda klasifikace genetika fyziologie MeSH
- chromozomy MeSH
- kapři parazitologie MeSH
- karyotypizace MeSH
- nemoci ryb parazitologie MeSH
- rozmnožování MeSH
- spermatocyty cytologie MeSH
- triploidie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The broad fish tapeworm Dibothriocephalus latus is a causative agent of human food-borne disease called diphyllobothriosis. Medical importance, scattered geographical distribution and unknown origin of D. latus in Europe and North America make this species to be an interesting model for population genetics. Microsatellite markers were originally designed by library screening using NGS approach and validated as tools for future studies on population genetics of D. latus. Out of 122 candidates selected after NGS analysis, 110 yielded PCR products of the expected size, and in 78 of them, a declared repetitive motif was confirmed by Sanger sequencing. After the fragment analysis, six loci were proved to be polymorphic and tested for observed (Ho) and expected (He) heterozygosity, and deviations from Hardy-Weinberg equilibrium (HWE). They promise future application in studies on genetic interrelationships, origin and migratory routes of this medically important emerging tapeworm.
- Klíčová slova
- Broad Fish tapeworm, Diphyllobothriidea, Microsatellites, Population genetics, Short tandem repeats,
- MeSH
- Diphyllobothrium klasifikace genetika MeSH
- genetická variace * MeSH
- genotypizační techniky metody MeSH
- mikrosatelitní repetice * MeSH
- populační genetika metody MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Evropa MeSH
- Severní Amerika MeSH
BACKGROUND: Population structure and genetic interrelationships of giant liver fluke Fascioloides magna from all enzootic North American regions were revealed in close relation with geographical distribution of its obligate definitive cervid hosts for the first time. METHODS: Variable fragments of the mitochondrial cytochrome c oxidase subunit I (cox1; 384 bp) and nicotinamide dehydrogenase subunit I (nad1; 405 bp) were applied as a tool. The concatenated data set of both cox1 and nad1 sequences (789 bp) contained 222 sequences that resulted in 50 haplotypes. Genetic data were analysed using Bayesian Inference (BI), Maximum Likelihood (ML) and Analysis of Molecular Variance (AMOVA). RESULTS: Phylogenetic analysis revealed two major clades of F. magna, which separated the parasite into western and eastern populations. Western populations included samples from Rocky Mountain trench (Alberta) and northern Pacific coast (British Columbia and Oregon), whereas, the eastern populations were represented by individuals from the Great Lakes region (Minnesota), Gulf coast, lower Mississippi, and southern Atlantic seaboard region (Mississippi, Louisiana, South Carolina, Georgia, Florida) and northern Quebec and Labrador. Haplotype network and results of AMOVA analysis confirmed explicit genetic separation of western and eastern populations of the parasite that suggests long term historical isolation of F. magna populations. CONCLUSION: The genetic makeup of the parasite's populations correlates with data on historical distribution of its hosts. Based on the mitochondrial data there are no signs of host specificity of F. magna adults towards any definitive host species; the detected haplotypes of giant liver fluke are shared amongst several host species in adjacent populations.
- MeSH
- Fasciola hepatica klasifikace enzymologie genetika izolace a purifikace MeSH
- fasciolóza epidemiologie parazitologie veterinární MeSH
- fylogeneze MeSH
- genetická variace MeSH
- molekulární sekvence - údaje MeSH
- přežvýkavci parazitologie MeSH
- proteiny červů genetika metabolismus MeSH
- respirační komplex IV genetika metabolismus MeSH
- vysoká zvěř MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- provincie Québec MeSH
- Spojené státy americké MeSH
- Názvy látek
- proteiny červů MeSH
- respirační komplex IV MeSH
