A proper soil risk assessment needs to estimate the processes that affect the fate and the behaviour of a contaminant, which are influenced by soil biotic and abiotic components. For this reason, the measurement of biomarkers in soil bioindicator organisms, such as earthworms, has recently received increasing attention. In this study, the earthworm Eisenia andrei was used to assess the pollutant-induced stress syndrome after exposure to sublethal concentrations of Cd (10 or 100 μg g(-1)) in OECD soil, after 14 d of exposure. Cadmium bioaccumulation and potential biomarkers such as catalase (CAT), hydrogen peroxide (H2O2), glutathione-S-transferase (GST), malondialdehyde (MDA), phenoloxidase (PO), metallothioneins (MTs) and genotoxic damage were determined. Results suggested that the exposure to 10 and 100 μg g(-1) Cd significantly increased Cd bioaccumulation, MTs and MDA; 100 μg g(-1) Cd contamination evidenced significantly higher values of H2O2 content and PO activity; CAT activity was inhibited at the higher concentration while GST and Comet assay did not show any significant differences from the control. Rank-based biomarker index showed that both different contaminated soils had an effect on the earthworms and allowed to validate the ecotoxicological relevance of this battery of biomarkers for a promising integrated multi-marker approach in soil monitoring and assessment.

• MeSH
• biologické markery analýza   metabolismus   MeSH
• ekotoxikologie metody   MeSH
• glutathiontransferasa analýza   metabolismus   MeSH
• kadmium metabolismus   toxicita   MeSH
• katalasa analýza   metabolismus   MeSH
• kometový test MeSH
• látky znečišťující půdu metabolismus   toxicita   MeSH
• malondialdehyd analýza   metabolismus   MeSH
• metalothionein analýza   metabolismus   MeSH
• monitorování životního prostředí metody   MeSH
• Oligochaeta účinky léků   genetika   metabolismus   MeSH
• peroxid vodíku analýza   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem 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.

• 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

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.

• 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

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

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.

• 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

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
• ferritin 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

Earthworms belonging to oligochaete annelids became a model for comparative immunologists in the early sixties with the publication of results from transplantation experiments that proved the existence of self/nonself recognition in earthworms. This initiated extensive studies on the earthworm immune mechanisms that evolved to prevent the invasion of pathogens. In the last four decades important cellular and humoral pathways were described and numerous biologically active compounds were characterized and often cloned.

• MeSH
• buněčná imunita imunologie   MeSH
• humorální imunita imunologie   MeSH
• Oligochaeta genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy 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
• práce podpořená grantem MeSH

Coelomic fluid of the Lumbricid Eisenia fetida contains a 42-kDa pattern recognition protein named coelomic cytolytic factor (CCF) that binds microbial cell wall components and triggers the activation of the prophenoloxidase cascade, an important invertebrate defense pathway. Here we report on the sequence characterization of CCF-like molecules of other Lumbricids: Aporrectodea caliginosa, Aporrectodea icterica, Aporrectodea longa, Aporrectodea rosea, Dendrobaena veneta, Lumbricus rubellus and Lumbricus terrestris, and show that CCF from E. fetida has a broader saccharide-binding specificity, being the only one recognizing N,N'-diacetylchitobiose. We suggest that the broad recognition repertoire of E. fetida CCF reflects a particular microbial environment this species lives in.

• MeSH
• cytotoxiny farmakologie   genetika   metabolismus   MeSH
• disacharidy metabolismus   MeSH
• financování organizované MeSH
• fylogeneze MeSH
• katecholoxidasa metabolismus   MeSH
• lektiny farmakologie   genetika   metabolismus   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• nádorové buněčné linie MeSH
• Oligochaeta genetika   metabolismus   MeSH
• prekurzory enzymů metabolismus   MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční seřazení MeSH
• substrátová specifita MeSH
• technika náhodné amplifikace polymorfní DNA MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• srovnávací studie MeSH

The coelomic fluid of the earthworm Eisenia fetida has been reported to contain a variety of proteins causing the lysis of red blood cells-EFAF (Eisenia fetida andrei factor), fetidin, lysenin, eiseniapore, and hemolysins isolated either from coelomic fluid (H1, H2, H3) or from cell lysate (CL(39) and CL(41)). We document the presence of two distinct genes with a high level of homology. These genes encode fetidin and lysenin but their level of expression differs in individual E. fetida andrei animals.

• MeSH
• biologické toxiny MeSH
• financování organizované MeSH
• hemolýza MeSH
• komplementární DNA genetika   MeSH
• molekulární sekvence - údaje MeSH
• Oligochaeta genetika   metabolismus   MeSH
• proteiny genetika   chemie   MeSH
• regulace genové exprese MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční homologie nukleových kyselin MeSH
• sekvenční seřazení MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH