Toll-like receptors (TLRs) play an important role in defense responses to pathogens in invertebrates. Here we characterize the first TLR isolated from an oligochaete annelid, namely, Eisenia andrei (EaTLR) and show its expression pattern. The full-length EaTLR cDNA consists of 2615 bp encoding a putative protein of 675 amino acids. The predicted amino acid sequence comprises of an extracellular domain containing 31 amino acid signal peptide and seven leucine-rich repeats (LRR), capped with cysteine-rich N- and C-terminal LRRs followed by a transmembrane domain and cytoplasmic Toll/IL-1R domain (TIR). TIR domains of twenty individual earthworms were sequenced and the variability suggesting the presence of a high number of TLR genes in the genome of E. andrei was observed. Phylogenetic analysis revealed the highest similarity of EaTLR with polychaete annelid, Capitella teleta and TLRs of mollusks and echinoderms. Finally, the highest constitutive expression of EaTLR was observed in the digestive tract. Gene expression was significantly increased in coelomocytes of E. andrei after the challenge with Gram-positive bacteria.

• Klíčová slova
• EaTLR, Eisenia andrei, Eisenia andrei Toll-like receptor, Innate immunity, Invertebrates, LRR, PAMPs, PRR, TLR, Toll-like receptor, leucine-rich repeat TIR, Toll/IL-1R domain, pathogen-associated molecular patterns, pattern recognition receptor,
• MeSH
• exprese genu MeSH
• fylogeneze MeSH
• klonování DNA metody   MeSH
• molekulární sekvence - údaje MeSH
• Oligochaeta genetika   metabolismus   MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční seřazení MeSH
• toll-like receptory biosyntéza   genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• toll-like receptory MeSH

Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) belong to the group of persistent organic pollutants, highly toxic environmental pollutants that include hydrophobic compounds with the tendency to bioaccumulate. Earthworms (Eisenia andrei) were exposed to PCDD/Fs-contaminated soil, and changes in their lipophilic structures and the gene expression of their defense molecules were followed. Damage to the intestinal wall and adjacent chloragogenous tissue was observed. Further, the up-regulation of the expression of several genes was detected. On the basis of these results, the mechanism of the impact of PCDD/Fs on earthworms has been proposed. Dioxins that accumulate in the lipophilic structures cause an increase in reactive oxidative species that triggers oxidative stress followed by the gene expression of two molecules that play a role in protection against oxidant toxicity, calreticulin (CRT) and Hsp70. Moreover, the effect of microbial biomass on the expression of coelomic cytolytic factor (CCF), a pattern recognition receptor, was also observed.

• Klíčová slova
• CCF, Calreticulin, Chloragogenous tissue, Eisenia andrei, Hsp70, PCDD/Fs, ROS,
• MeSH
• benzofurany analýza   metabolismus   MeSH
• dioxiny analýza   metabolismus   MeSH
• látky znečišťující půdu analýza   metabolismus   MeSH
• Oligochaeta účinky léků   genetika   fyziologie   MeSH
• půda chemie   MeSH
• regulace genové exprese účinky léků   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• benzofurany MeSH
• dibenzo(1,4)dioxin MeSH Prohlížeč
• dibenzofuran MeSH Prohlížeč
• dioxiny MeSH
• látky znečišťující půdu MeSH
• půda MeSH

LBP/BPIs are pattern recognition receptors that are often present in vertebrates and in invertebrates, and they play a defense role against pathogens. We have identified 1698 bp cDNA sequence from the Eisenia andrei earthworm with predicted amino acid sequence that shares homology with the LBP/BPI family (EaLBP/BPI). Sequence analysis of EaLBP/BPI proved the existence of two conserved domains with the potential ability to bind LPS. The predicted molecular mass of the EaLBP/BPI protein is 53.5 kDa, and its high basicity (pI 9.8) is caused by its high arginine content. Constitutive transcription of the Ealbp/bpi gene was shown in all tested tissues, with the highest level in coelomocytes and seminal vesicles; the lowest level was detected in the intestine. On the contrary, another earthworm LPS-binding molecule CCF (coelomic cytolytic factor) was expressed only in the intestine and coelomocytes. In E. andrei coelomocytes, the transcription of Ealbp/bpi gene was up-regulated in response to bacterial stimulation, reaching a maximum at 8 and 16 h post stimulation with Bacillus subtilis and Escherichia coli, respectively.

• Klíčová slova
• CCF, Earthworm, Eisenia, LBP/BPI, LPS, PRR,
• MeSH
• fylogeneze MeSH
• hybridizace in situ MeSH
• kationické antimikrobiální peptidy genetika   MeSH
• klonování DNA MeSH
• krevní proteiny genetika   MeSH
• molekulární sekvence - údaje MeSH
• Oligochaeta genetika   imunologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bactericidal permeability increasing protein MeSH Prohlížeč
• kationické antimikrobiální peptidy MeSH
• krevní proteiny MeSH

Agriculture is today indispensably connected with enormous use of pesticides. Despite tough regulation, their entrance into soil cannot be excluded and they might enter soil organisms and plants and continue further to terrestrial food chains. This study was conducted to investigate the bioaccumulation of two pesticides currently used in large amounts, the insecticide chlorpyrifos (CLP) and the fungicide tebuconazole (TBZ). Their detailed uptake kinetics in the model earthworm species Eisenia andrei were measured in two arable soils differing in organic carbon content (1.02 and 1.93% respectively). According to our results, a steady state was reached after 3-5 days for both pesticides and soils. The values of bioaccumulation factors calculated at the steady state ranged from 4.5 to 6.3 for CLP and 2.2-13.1 for TBZ. Bioaccumulation factors were also calculated as the ratio of uptake and elimination rate constants with results comparable with steady-state bioaccumulation factors. The results suggested that the degradation and bioaccumulation of tested compounds might be influenced by other factors than only total organic carbon (e.g. clay content). The lower Koc and hydrophobicity of TBZ relative to CLP probably led to higher availability of TBZ through pore water exposure. On the other hand, CLP's higher hydrophobicity probably caused an increase in availability by its additional uptake via ingestion. To enable a proper ecological risk assessment of current pesticides in soils, it is necessary to accurately determine their bioaccumulation in soil invertebrates. We believe that our study not only brings such information for two specific pesticides but also addresses key methodological issues in this area.

• Klíčová slova
• Bioaccumulation, Chlorpyrifos, Currently used pesticides, Eisenia andrei, Soil, Tebuconazole, Toxicokinetics,
• MeSH
• dursban analýza   metabolismus   MeSH
• jíl MeSH
• kinetika MeSH
• látky znečišťující půdu analýza   metabolismus   MeSH
• monitorování životního prostředí MeSH
• Oligochaeta metabolismus   MeSH
• pesticidy analýza   metabolismus   MeSH
• půda chemie   MeSH
• silikáty hliníku analýza   MeSH
• triazoly analýza   metabolismus   MeSH
• zemědělství MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• dursban MeSH
• jíl MeSH
• látky znečišťující půdu MeSH
• pesticidy MeSH
• půda MeSH
• silikáty hliníku MeSH
• tebuconazole MeSH Prohlížeč
• triazoly MeSH

Earthworms Eisenia andrei, similarly to other invertebrates, rely on innate defense mechanisms based on the capability to recognize and respond to nonself. Here, we show a correlation between the expression of CCF, a crucial pattern-recognition receptor, and lysozyme, with enzyme activities in the gut of E. andrei earthworms following a microbial challenge. These data suggest that enzyme activities important for the release and recognition of molecular patterns by pattern-recognition molecules, as well as enzymes involved in effector pathways, are modulated during the microbial challenge. In particular, protease, laminarinase, and glucosaminidase activities were increased in parallel to up-regulated CCF and lysozyme expression.

• Klíčová slova
• Digestive enzyme, Earthworm, Eisenia andrei, Gut, Innate immunity, Microbiota,
• MeSH
• Bacillus subtilis imunologie   MeSH
• celulasy metabolismus   MeSH
• Escherichia coli imunologie   MeSH
• hexosaminidasy metabolismus   MeSH
• messenger RNA metabolismus   MeSH
• muramidasa metabolismus   fyziologie   MeSH
• Oligochaeta enzymologie   imunologie   mikrobiologie   MeSH
• přirozená imunita MeSH
• proteasy metabolismus   MeSH
• Saccharomyces cerevisiae imunologie   MeSH
• upregulace MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• celulasy MeSH
• hexosaminidasy MeSH
• messenger RNA MeSH
• muramidasa MeSH
• proteasy MeSH

The massive production and use of silver nanoparticles (Ag NPs) have led to their increasing release into the environment. Even though the antimicrobial and cytotoxic effects of native nanoparticles have been well studied, the environmental impacts of transformation products such as silver sulfide nanoparticles (Ag2S NPs) have not been elucidated. In the present study, we assessed the toxicity of Ag2S NPs and silver nitrate (AgNO3), as a source of Ag, to the earthworm Eisenia andrei using a nominal concentration of 5 mg Ag kg-1 soil. We used the OECD guidelines to assess effects on weight loss and mortality for 14 days. After exposure, we also extracted the immune effector cells (coelomocytes) and conducted a battery of biomarker tests. To ensure the quality of the toxicological results, the structural changes of NPs during the experiment and the uptake of silver by the earthworms were monitored. During the experiment, mortality effects were not detected, but a weight loss was observed in the earthworms exposed to Ag2S NPs. Altough Ag2S NPs were engulfed by E. andrei cells, neither phenoloxidase activity nor lipid peroxidation differed from the untreated control group. Cells from earthworms treated with Ag2S NPs exerted very broad value range of nitric oxide (NO) generation, suggesting an imbalance in the NO metabolism. Overall, this study suggests minimal risks associated with Ag2S NPs exposure to earthworms. However, further studies are needed to assure no immunotoxicological or chronic effects on a wider range of terrestrial organisms.

• Klíčová slova
• Coelomocytes, Earthworm, Lipid peroxidation, Nanoparticles, Phenoloxidase,
• MeSH
• dusičnan stříbrný toxicita   MeSH
• hmotnostní úbytek MeSH
• kovové nanočástice * chemie   toxicita   MeSH
• látky znečišťující půdu * toxicita   MeSH
• Oligochaeta * MeSH
• půda chemie   MeSH
• sloučeniny stříbra MeSH
• stříbro metabolismus   toxicita   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• dusičnan stříbrný MeSH
• látky znečišťující půdu * MeSH
• půda MeSH
• silver sulfide MeSH Prohlížeč
• sloučeniny stříbra MeSH
• stříbro MeSH

One 25-kg dog produces about 500 g of excrement per day. Excrement is a potentially hazardous material, as it may contain pathogenic microorganisms. Our samples were tested for the presence of thermotolerant coliform bacteria, Enterococcus spp., Escherichia coli and Salmonella spp., which are indicators of faecal contamination, as well as for the presence of helminths and their eggs. During the experiment, it was observed whether these microorganisms could be eliminated by vermicomposting. There were two variants of vermicomposting piles: one test pile (with continuous feeding) and one control pile (with a single feeding). The vermicomposting process was run in outdoor conditions in park for 51 weeks using Eisenia andrei earthworms. The vermicomposting of dog excrement with waste from park maintenance (1:2) can produce a good quality fertiliser. During the process of vermicomposting, there was a gradual decrease in the content of pathogenic bacteria. At the end of the vermicomposting process, there were no eggs or adult helminths. The vermicompost was very rich in microorganisms and enzymatic activity. The pH value was slightly alkaline, and the C:N ratio corresponded to value of mature vermicompost.

• Klíčová slova
• PLFA analyses, Vermicomposting, dog excrement, endoparasites, enzymatic activity, pathogenic organisms,
• MeSH
• Oligochaeta * MeSH
• průmyslová hnojiva MeSH
• psi MeSH
• půda chemie   MeSH
• zvířata MeSH
• Check Tag
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• průmyslová hnojiva MeSH
• půda MeSH

Survival of earthworms in the environment depends on their ability to recognize and eliminate potential pathogens. This work is aimed to compare the innate defense mechanisms of two closely related earthworm species, Eisenia andrei and Eisenia fetida, that inhabit substantially different ecological niches. While E. andrei lives in a compost and manure, E. fetida can be found in the litter layer in forests. Therefore, the influence of environment-specific microbiota on the immune response of both species was followed. Firstly, a reliable method to discern between E. andrei and E. fetida based on species-specific primers for cytochrome c oxidase I (COI) and stringent PCR conditions was developed. Secondly, to analyze the immunological profile in both earthworm species, the activity and expression of lysozyme, pattern recognition protein CCF, and antimicrobial proteins with hemolytic function, fetidin and lysenins, have been assessed. Whereas, CCF and lysozyme showed only slight differences in the expression and activity, fetidin/lysenins expression as well as the hemolytic activity was considerably higher in E. andrei as compared to E. fetida. The expression of fetidin/lysenins in E. fetida was not affected upon the challenge with compost microbiota, suggesting more substantial changes in the regulation of the gene expression. Genomic DNA analyses revealed significantly higher level of fetidin/lysenins (determined using universal primer pairs) in E. andrei compared to E. fetida. It can be hypothesized that E. andrei colonizing compost as a new habitat acquired an evolutionary selection advantage resulting in a higher expression of antimicrobial proteins.

• MeSH
• Bacteria klasifikace   genetika   imunologie   MeSH
• biologické toxiny genetika   imunologie   MeSH
• cytotoxicita imunologická genetika   imunologie   MeSH
• druhová specificita MeSH
• ekosystém * MeSH
• exprese genu MeSH
• hemolýza genetika   imunologie   MeSH
• hnůj mikrobiologie   parazitologie   MeSH
• molekulární sekvence - údaje MeSH
• muramidasa genetika   imunologie   MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• Oligochaeta klasifikace   genetika   imunologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• přirozená imunita genetika   imunologie   MeSH
• proteiny genetika   imunologie   MeSH
• půdní mikrobiologie MeSH
• respirační komplex IV genetika   MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie nukleových kyselin MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické toxiny MeSH
• fetidin MeSH Prohlížeč
• hnůj MeSH
• lysenin MeSH Prohlížeč
• muramidasa MeSH
• proteiny MeSH
• respirační komplex IV MeSH

Lysozyme is a widely distributed antimicrobial protein having specificity for cleaving the beta-(1,4)-glycosidic bond between N-acetylmuramic acid (NAM) and N-acetylglucosamine (GlcNAc) of peptidoglycan of the bacterial cell walls and thus efficiently contributes to protection against infections caused mainly by Gram-positive bacteria. In the present study, we assembled a full-length cDNA of a novel invertebrate-type lysozyme from Eisenia andrei earthworm (EALys) by RT-PCR and RACE system. The primary structure of EALys shares high homology with other invertebrate lysozymes; however the highest, 72% identity, was shown for the destabilase I isolated from medicinal leech. Recombinant EALys expressed in Escherichia coli exhibited the lysozyme and isopeptidase activity. Moreover, real-time PCR revealed increased levels of lysozyme mRNA in coelomocytes of E. andrei after the challenge with both Gram-positive and Gram-negative bacteria.

• MeSH
• Bacillus subtilis imunologie   patogenita   MeSH
• bakteriální adheze MeSH
• chitinasy metabolismus   MeSH
• Echinodermata genetika   MeSH
• endopeptidasy metabolismus   MeSH
• Escherichia coli genetika   imunologie   patogenita   MeSH
• glukosamin analogy a deriváty   imunologie   metabolismus   MeSH
• grampozitivní bakteriální infekce imunologie   MeSH
• hydrolýza MeSH
• infekce vyvolané Escherichia coli imunologie   MeSH
• interakce hostitele a patogenu MeSH
• klonování DNA MeSH
• kyseliny muramové imunologie   metabolismus   MeSH
• lyasy štěpící vazby C-N metabolismus   MeSH
• muramidasa genetika   imunologie   metabolismus   MeSH
• Oligochaeta enzymologie   genetika   imunologie   MeSH
• pijavka lékařská genetika   MeSH
• sekvenční homologie MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chitinasy MeSH
• endopeptidasy MeSH
• fibrin destabilase MeSH Prohlížeč
• glukosamin MeSH
• isopeptidase MeSH Prohlížeč
• kyseliny muramové MeSH
• lyasy štěpící vazby C-N MeSH
• muramidasa MeSH
• N-acetylglucopyranosylamine MeSH Prohlížeč
• N-acetylmuramic acid MeSH Prohlížeč

Ferritin is a storage protein that plays a key role in iron metabolism. In this study, we report on the sequence characterization of a ferritin-coding cDNA in Eisenia andrei earthworms isolated by RT-PCR using degenerated primers, and we suggest the presence of a putative IRE in the 5'-UTR of ferritin mRNA. The obtained ferritin sequence was compared with those of other animals showing sequence and structure homology in consensus sites, including the iron-responsive element (IRE) and ferroxidase centers. Despite the sequence homology in the E. andrei mRNA of ferritin with the sequences of other animals in consensus IRE sites, the presented cytosine in the IRE of E. andrei ferritin in the expected position does not form a conventional bulge. The presence of ferritin in the coelomic fluid of E. andrei was proven by iron staining assay. Moreover, aconitase activity in the coelomic fluid was assessed by aconitase assay, suggesting the presence of an iron regulatory protein. Quantitative analysis revealed changes in the gene expression levels of ferritin in coelomocytes in response to bacterial challenge, reaching the maximum level 8h after the stimulation with both Gram-positive and Gram-negative bacteria.

• MeSH
• akonitáthydratasa analýza   genetika   metabolismus   MeSH
• Bacillus subtilis metabolismus   MeSH
• ceruloplasmin genetika   MeSH
• DNA primery MeSH
• Escherichia coli K12 metabolismus   MeSH
• ferritiny chemie   genetika   MeSH
• klonování DNA MeSH
• komplementární DNA biosyntéza   genetika   izolace a purifikace   MeSH
• konformace nukleové kyseliny MeSH
• messenger RNA chemie   genetika   MeSH
• molekulární sekvence - údaje MeSH
• Oligochaeta genetika   metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• regulace genové exprese MeSH
• RNA genetika   izolace a purifikace   MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• akonitáthydratasa MeSH
• ceruloplasmin MeSH
• DNA primery MeSH
• ferritiny MeSH
• komplementární DNA MeSH
• messenger RNA MeSH
• RNA MeSH