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1.
Chemosphere ; 240: 124902, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31563721

RESUMO

Eisenia fetida earthworm is an ecotoxicologically important test species to monitor various pollutants. However, there is a little knowledge about the effects of cadmium (Cd) on earthworms at the transcriptional level. Firstly, we exposed E. fetida to soils supplemented with different concentrations (10, 30, 60 mg/kg soil) of Cd. Moreover, we depicted the characterization of gene expressions with E. fetida using high-throughput profiling of gene expression. In addition, a comparison of the gene expression profiles between each Cd treatment group and the control group suggested that differential expressional genes (DEGs) mainly enriched in enzyme activity, metabolism, oxidative stress, regeneration and apoptosis pathways. 8 DEGs from these pathways had been selected randomly to confirm the data of RNA-seq. Among these DEGs, six genes (metallothionein-2, phytochelatin synthase 1a, CuZn superoxide dismutase, sex determining region Y-box 2, sex determining region Y-box 4b, TP53-regulated inhibitor of apoptosis 1-like) up-regulated and 2 genes (beta-1,4-endoglucanase, apoptosis-stimulating of p53 protein 2-like) down-regulated in response to Cd exposure. The alteration of them indicated that earthworms could reduce the toxicity and bioavailability of Cd in polluted soil ecosystems through different pathways. This work lays an important foundation for linking earthworm transcriptional level with the ecological risk of Cd in soil ecosystem.


Assuntos
Cádmio/toxicidade , Oligoquetos/fisiologia , Poluentes do Solo/toxicidade , Aminoaciltransferases , Animais , Disponibilidade Biológica , Cádmio/análise , Ecossistema , Perfilação da Expressão Gênica , Metalotioneína/genética , Metalotioneína/metabolismo , Oligoquetos/efeitos dos fármacos , Oligoquetos/genética , Estresse Oxidativo/efeitos dos fármacos , Solo , Poluentes do Solo/análise , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo
2.
Parasitol Res ; 118(12): 3359-3370, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31729575

RESUMO

A new species of naidid oligochaete, Dero rwandae, detected in the bladder and the Wolffian ducts of reed frogs Hyperolius kivuensis from Rwanda, is described. Until now, D. bauchiensis was the only endoparasitic Dero known to infect African frogs infesting the eyes and Harderian glands. To the best of our knowledge, the finding of D. rwandae is the first record of an African Dero species infecting the urinary tract of anurans. In general morphology, the two African Dero parasites resemble each other, but differences in the features of ventral setae morphology exist. Parts of the mitochondrial 16S rRNA locus and the nuclear 18S and 28S rRNA loci were sequenced to assess the phylogenetic relationships to other Dero spp. Among those few species, that are barcoded so far, the closest relative of the new taxon is D. superterrenus, a free-living South American species. The species groups formerly termed subgenera Allodero, Aulophorus and Dero within the genus Dero do not represent distinct evolutionary lineages and the genus is paraphyletic including Branchiodrilus.


Assuntos
Anuros/parasitologia , Oligoquetos/classificação , Doenças Parasitárias em Animais/parasitologia , Bexiga Urinária/parasitologia , Animais , Sequência de Bases , Oligoquetos/genética , Filogenia , RNA Ribossômico/genética , RNA Ribossômico 16S/genética , RNA Ribossômico 28S/genética , Ruanda
3.
Environ Int ; 129: 10-17, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31102950

RESUMO

Phthalates (phthalic acid esters) have been widely applied as plasticizers. They are ubiquitous contaminants in soils, thereby posing a threat to human health. In this study, ecotoxicological effects of three typical PAEs (dimethyl phthalate-DMP, di-n-octyl phthalate-DOP and butyl benzyl phthalate-BBP) were investigated. As a biological indicator, earthworms (Eisenia fetida) were exposed to phthalates at various doses (0, 0.1, 1, 10 and 50 mg/kg) for different times (7, 14, 21, and 28 d). We evaluated the effects of phthalates on reactive oxygen species (ROS) generation, antioxidant enzymes (superoxide dismutase-SOD, peroxidase-POD and catalase-CAT) activities, glutathione S-transferase enzyme (GST) activity, malondialdehyde (MDA) content and DNA damage. Results showed that ROS content increased with increasing phthalates, whereas ROS content generally increased and then decreased with exposure time. However, antioxidant enzymes activities in earthworms displayed different trends. The GST activity in high-dose treatment group was significantly activated. For DMP and DOP, lipid peroxidation mainly occurred between 14 and 28 d, while for BBP, it primarily existed after 7 d and then disappeared after 28 d. Besides, comet assay indicated that there was a dose-response relationship between the DNA damage and phthalate dose, following DMP > DOP > BBP. Given their toxicity, it is important to understand the mechanisms associated with their eco-toxicity and to reduce their adverse impacts on the environment.


Assuntos
Oligoquetos/efeitos dos fármacos , Ácidos Ftálicos/toxicidade , Poluentes do Solo/toxicidade , Animais , Antioxidantes/metabolismo , Dano ao DNA , Malondialdeído/metabolismo , Oligoquetos/enzimologia , Oligoquetos/genética , Oligoquetos/fisiologia , Estresse Oxidativo/efeitos dos fármacos
4.
Environ Pollut ; 251: 453-459, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31103005

RESUMO

Microplastic (MP) pollution is everywhere. In terrestrial environments, microfibres (MFs) generated from textile laundering are believed to form a significant component of MPs entering soils, mainly through sewage sludge and compost applications. The aim of this study was to assess the effect of MFs on a keystone soil organism. We exposed the earthworm Lumbricus terrestris to soil with polyester MFs incorporated at rates of 0, 0.1 and 1.0 %w/w MF for a period of 35 days (in the dark at 15 °C; n = 4 for each treatment). Dried plant litter was applied at the soil surface as a food source for the earthworms. We assessed earthworm vitality through mortality, weight change, depurate production and MF avoidance testing. In addition, we measured stress biomarker responses via the expression of metallothionein-2 (mt-2), heat shock protein (hsp70) and superoxide dismutase (sod-1). Our results showed that exposure and ingestion of MFs (as evidenced by subsequent retrieval of MFs within earthworm depurates) were not lethal to earthworms, nor did earthworms actively avoid MFs. However, earthworms in the MF1.0% treatment showed a 1.5-fold lower cast production, a 24.3-fold increase in expression of mt-2 (p < 0.001) and a 9.9-fold decline in hsp70 expression (p < 0.001). Further analysis of soil and MF samples indicated that metal content was not a contributor to the biomarker results. Given that burrowing and feeding behaviour, as well as molecular genetic biomarkers, were modulated in earthworms exposed to MFs, our study highlights potential implications for soil ecosystem processes due to MF contamination.


Assuntos
Oligoquetos/efeitos dos fármacos , Plásticos/toxicidade , Poliésteres/toxicidade , Poluentes do Solo/toxicidade , Animais , Comportamento Animal/efeitos dos fármacos , Biomarcadores Ambientais/efeitos dos fármacos , Biomarcadores Ambientais/genética , Oligoquetos/genética , Oligoquetos/metabolismo , Plásticos/metabolismo , Poliésteres/metabolismo , Poluentes do Solo/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética
5.
Chemosphere ; 227: 570-579, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31004823

RESUMO

In this study, the acute and subchronic toxicity of 1-alkyl-3-methyl imidazole bromide ionic liquids (ILs) [Cnmim]Br (n = 2, 4, 6, 10, 12) was evaluated utilizing earthworms (Eisenia fetida) as a basis for their impact on terrestrial ecosystems. The filter paper tests and artificial soil tests were conducted as acute toxicity tests to investigate the LC50 of ILs, while in subchronic toxicity tests, earthworms were exposed to ILs in artificial soil (5, 10, 20 and 40 mg kg-1) for 28 d. Reactive oxygen species (ROS), antioxidant enzymes, detoxifying enzymes and oxidative damage were measured to determine subchronic effects of ILs on E. fetida. The results showed that when the earthworms were exposed to these five ILs in acute toxicity experiments, [C2mim]Br had the lowest toxicity, as the alkyl length increased, the toxicity increased up to [C10mim]Br: a "cut-off effect" (decreased toxicity) was observed at [C12mim]Br. The results highlight the varying toxicity of ILs with different alkyl chains to E. fetida and provide valuable data for detailing the impact of ILs on ecological receptors.


Assuntos
Dano ao DNA , Hidrocarbonetos Bromados/toxicidade , Imidazóis/toxicidade , Líquidos Iônicos/toxicidade , Oligoquetos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Hidrocarbonetos Bromados/química , Imidazóis/química , Inativação Metabólica , Líquidos Iônicos/química , Dose Letal Mediana , Oligoquetos/genética , Oligoquetos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Testes de Toxicidade Aguda
6.
Chemosphere ; 226: 47-59, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30913427

RESUMO

Exogenous retinoic acid (RA) delays and disturbs the regeneration of Eisenia foetida and inhibits the expression of pluripotent gene Sox2. However, studies of E. foetida conducted at the molecular level have been unable to elucidate its regeneration and mechanisms of RA effects on its regeneration. We merged existing transcriptomic data for E. foetida to generate a high-confidence set of transcriptomes. The de novo assembly of transcriptomes was performed by using the Trinity method, and functional annotations were analysed. We performed RNA-seq on four samples of regenerating tail fragments, three across a time-course (0, 3 and 7 days post amputation) and the fourth sample exposed to RA (7 days post amputation). E. foetida regeneration genes underwent significant upregulation and downregulation over the examined time periods, which may have been caused by a shared regulatory programme controlled by multiple gene families. The inhibition of RA against earthworm regeneration is likely related to the expression of these genes. Using annotation data and clustering, we also identified specific transcripts of 6 gene superfamilies enriched among genes exhibiting differential expression during regeneration periods and exhibiting the same expression patterns as those of the Sox2 gene. The regeneration transcriptome of tail fragment regeneration serves as a strong resource for investigating global expression changes that occur during regeneration and the toxicity of RA. This study offers insight for better understanding the regeneration of lower animals and molecular mechanisms of RA toxicity in invertebrates.


Assuntos
Oligoquetos/genética , Transcriptoma/genética , Tretinoína/uso terapêutico , Animais , Regeneração , Tretinoína/farmacologia
7.
BMC Evol Biol ; 19(Suppl 1): 50, 2019 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-30813890

RESUMO

BACKGROUND: Many earthworm species demonstrate significant cryptic diversity, with several highly diverged mitochondrial lineages found within most of the taxa studied to date. The status of differences between these lineages on the nuclear level is still unclear. Because of widespread polyploidy in earthworms, most studies were limited to two nuclear loci, the ribosomal and the histone clusters. Here we attempted to elucidate the status of a set of genetic lineages within Eisenia nordenskioldi nordenskioldi, an earthworm species from Northern Asia with high intraspecific diversity. We performed RNA-seq on an IonTorrent platform for five specimens of this species belonging to five genetic lineages, as well as two outgroups from the family Lumbricidae, the congenetic E. andrei, and Lumbricus rubellus. RESULTS: We de novo assembled transcriptomes and constructed datasets of genes present in all seven specimens using broad (ProteinOrtho; 809 genes) and narrow (HaMStR; 203 genes) ortholog assignment. The majority of orthologs had identical amino acid sequences in all studied specimens, which we believe was due to strong bias towards the most conserved genes. However, for the rest of genes the differences among the lineages were lower than those between them and the congeneric E. andrei. Both datasets yielded phylogenetic trees with the same topology. E. n. nordenskioldi was found to be monophyletic. The differences on the genetic level had no concordance with geography, implying complex history of dispersal. CONCLUSIONS: We found that genetic lineages of E. n. nordenskioldi are genetically distinct on nuclear level and probably diverged long ago. Current data implies that they might even represent distinct species within the E. nordenskioldi species complex.


Assuntos
Núcleo Celular/genética , Perfilação da Expressão Gênica , Genômica , Oligoquetos/citologia , Oligoquetos/genética , Simpatria , Animais , DNA Mitocondrial/genética , Filogenia
8.
Biosci Rep ; 39(2)2019 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-30705087

RESUMO

Guang-dilong (Pheretima aspergillum) is a traditional Chinese animal medicine that has been used for thousands of years in China. In the present study, we purposed to establish a new rapid identification method for Guang-dilong. We provided a useful technique, loop-mediated isothermal amplification (LAMP), to differentiate Guang-dilong from other species. Four specific LAMP primers were designed based on mitochondrial cytochrome c oxidase I (COI) gene sequences of Guang-dilong. LAMP reaction, containing DNA template, four primers, 10× Bst DNA polymerase reaction buffer, dNTPs, MgSO4, and Bst DNA polymerase, was completed within 60 min at 63°C. The LAMP product can be visualized by adding SYBR Green I or detected by 2% gel electrophoresis. LAMP technology was successfully established for rapid identification of Guang-dilong. In addition, DNA template concentration of 675 fg/µl was the detection limit of LAMP in Guang-dilong, which was 1000-times higher than conventional PCR. The simple, sensitive, and convenient LAMP technique is really suited for on-site identification of Guang-dilong in herbal markets.


Assuntos
Oligoquetos/genética , Reação em Cadeia da Polimerase/métodos , Animais , DNA/genética , Primers do DNA/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Genes Mitocondriais , Oligoquetos/classificação , Reação em Cadeia da Polimerase/economia
9.
Mol Med Rep ; 19(4): 2537-2550, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30720084

RESUMO

Earthworms are useful indicator organisms of soil health and Eisenia fetida have been extensively used as test organisms in ecotoxicological studies. In order to gain insight into the gene expression profiles associated with physiological functions of earthworms, a full­length enriched cDNA library of the Eisenia fetida genome was successfully constructed using Switching Mechanism at 5'End of RNA Template technology. Construction of a cDNA library and analysis of Expressed Sequence Tags (ESTs) are efficient approaches for collecting genomic information and identifying genes important for a given biological process. Furthermore, analysis of the expression abundance of ESTs was performed with the aim of providing genetic and transcriptomic information on the development and regenerative process of earthworms. Phrep and Crossmatch were used to process EST data and a total of 1,140 high­quality EST sequences were determined by sequencing random cDNA clones from the library. Clustering analysis of sequences revealed a total of 593 unique sequences including 225 contiguous and 368 singleton sequences. Basic Local Alignment Search Tool analysis against the Kyoto Encyclopedia of Genes and Genomes database resulted in 593 significant hits (P­value <1x10­8), of which 168 were annotated through Gene Ontology analysis. The STRING database was used to determine relationships among the 168 ESTs, identifying associated genes involved in protein­protein interactions and gene expression regulation. Based on nucleic acid and protein sequence homology, the mutual relationships between 287 genes could be obtained, which identified a portion of the ESTs as known genes. The present study reports on the construction of a high­quality cDNA library representative of adult earthworms, on a preliminary analysis of ESTs and on a putative functional analysis of ESTs. The present study is expected to enhance our understanding of the molecular basis underlying the biological development of earthworms.


Assuntos
Etiquetas de Sequências Expressas , Biblioteca Gênica , Oligoquetos/genética , Análise de Sequência de DNA , Animais , Biologia Computacional/métodos , Ontologia Genética , Anotação de Sequência Molecular
10.
PLoS One ; 14(1): e0208904, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30699114

RESUMO

Lumbricidae taxonomy is vastly restricted by the morphological simplicity of earthworms and their lack of complex appendices. This has led to confusing results in the Lumbricidae classifications, which in turn, has hindered our ability to identify and assign new and cryptic species to the family. Here we propose the addition of a new Lumbricidae genus from the Zagros and Elburz Mountains of Iran, i.e. Philomontanus gen. nov, including three new species. Our taxonomic inferences were based on the phylogenetic analysis of two nuclear gene regions (28S rDNA and 18S rDNA) and 11 mitochondrial gene regions (16S rDNA, 12S rDNA, NADH dehydrogenase I, cytochrome oxidase subunits I and II and tRNAs Asn, Asp, Val, Leu, Ala and Ser). Philomontanus gen. nov comprises the earthworm species Philomontanus sarii sp. nov., Philomontanus mahmoudi sp. nov. and Philomontanus baloutchi sp. nov. These three species are morphologically similar to each other with only a few characters separating them (e.g. size, pigmentation and position of clitellum). Our findings support the adoption of an integrative approach including molecular information (e.g., DNA sequences) to aid earthworm classification and develop a robust taxonomy.


Assuntos
Oligoquetos/genética , Animais , DNA Ribossômico/genética , Complexo I de Transporte de Elétrons/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Evolução Molecular , Oligoquetos/classificação , Filogenia
11.
Genes Genomics ; 41(3): 367-371, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30610619

RESUMO

BACKGROUND: The invertebrate type (i-type) lysozyme not showing a clear homology with the known types of lysozyme was first demonstrated from a marine bivalve, conch and earthworm by N-terminal sequence. An i-type lysozyme isolated from the earthworm found to be up-regulated upon bacterial challenge, suggesting this lysozyme to function as an inducible immune factor. However, information on the i-type lysozyme related with digestive function is very limited in the earthworm. OBJECTIVE: The objective of this study is to investigate the molecular characteristics and function of the new i-type lysozyme from the earthworm. METHODS: To identify a new i-type lysozyme, multiple amino acid sequence alignment and phylogenetic analyses were employed. Its mRNA expression pattern was observed by fluorescent in situ hybridization (FISH). RESULTS: A new i-type lysozyme (Ea-iLys) from an earthworm, Eisenia andrei with the open reading frame of 678 bp (226 amino acid residues) appeared to comprise conserved 14 cysteine residues for disulfide bridges and amino acid residues for the enzyme activities of lysozyme and isopeptidase, of which mRNA expression is mainly localized in the lining of midgut epithelium. No significant expression signal was detected in immune competent sites such as chloragogue tissue, typhlosole region, body coelom and muscle layers. CONCLUSION: Our results suggest that this enzyme primarily acts as a digestive enzyme rather than an innate immune factor.


Assuntos
Muramidase/genética , Oligoquetos/enzimologia , Animais , Mucosa Intestinal/metabolismo , Muramidase/química , Muramidase/metabolismo , Músculo Esquelético/metabolismo , Oligoquetos/genética
12.
Environ Sci Pollut Res Int ; 26(3): 2851-2863, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30499083

RESUMO

In order to ensure better use of treated wastewater (TWW), we investigated the effect of three increasing doses of TWW, 10%, 50%, and 100%, on biochemical and transcriptomic statuses of earthworms Eisenia andrei exposed during 7 and 14 days. The effect of TWW on the oxidative status of E. andrei was observed, but this effect was widely dependent on the dilution degree of TWW. Results showed a significant decrease in the catalase (CAT) activity and an increase in the glutathione-S-transferase (GST) activity, and considerable acetylcholinesterase (AChE) inhibition was recorded after 14 days of exposure. Moreover, malondialdehyde (MDA) accumulation was found to be higher in exposed animals compared to control worms. The gene expression level revealed a significant upregulation of target genes (CAT and GST) during experimentation. These data provided new information about the reuse of TWW and its potential toxicity on soil organisms.


Assuntos
Irrigação Agrícola/métodos , Expressão Gênica/efeitos dos fármacos , Oligoquetos/efeitos dos fármacos , Poluentes do Solo/toxicidade , Solo/química , Águas Residuárias/toxicidade , Animais , Catalase/genética , Catalase/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Malondialdeído/metabolismo , Oligoquetos/enzimologia , Oligoquetos/genética , Poluentes do Solo/química , Águas Residuárias/química
13.
Sci Total Environ ; 651(Pt 1): 1236-1242, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30360255

RESUMO

Persistent Organic Pollutants (POPs) are stable organic chemicals that represent a potential risk for ecosystems due to their high toxicity, persistence and biomagnification through food chains. Bioindicators in ecosystems have emerged to assess the effect of environmental pollutants. Earthworms are some of the most common bioindicator organisms in terrestrial ecosystems. The main objective of this study was to evaluate the geontoxicity of POP exposure in wild earthworms captured at different levels of urbanization throughout the lower basin of the Coatzacoalcos River (industrial, urban and rural areas). POP soil and earthworm tissue concentrations were measured via Gas-Mass Chromatography, and earthworm DNA damage was evaluated through the comet assay. The greatest concentrations of ΣPOPs, DDT and HCH were found in soil from industrial sites, followed by urban and rural areas (504.68, 383.10, 298.16; 22.6, 4.6, 2.6 and 433.7, 364, 255.6 mg/kg, respectively). Unlike other pollutants, mean ΣPCBs values were highest for industrial soil samples, followed by those from rural and urban areas (41.10, 33.97 and 12.44 mg/kg respectively). For all earthworm tissue POP analyses, the highest concentrations were found in individuals from industrial sites, followed by the urban and rural areas. Furthermore, the highest levels of DNA damage were registered in the industrial area, followed by the urban and rural areas. These assays suggest a strong links among regional soil contamination, POPs bioavailability and the potential risk of detrimental health effects for organisms that inhabit surface soil (soil life). Earthworms contribute vital ecosystem services that could be affected by these results. This work provides evidence of the potential ecological risk that exists in the Lower Basin of the Coatzacoalcos River.


Assuntos
Dano ao DNA , Exposição Ambiental , Oligoquetos/efeitos dos fármacos , Poluentes do Solo/efeitos adversos , Animais , Monitoramento Ambiental , México , Oligoquetos/genética
14.
Dev Comp Immunol ; 90: 41-46, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30179632

RESUMO

Lumbricin and its orthologue antimicrobial peptides were typically isolated from annelids. In this report, mRNA for lumbricin and -serendipitously- a novel lumbricin-related mRNA sequence were identified in Eisenia andrei earthworms. The determined mRNA sequences of E. andrei lumbricin and lumbricin-related peptide consist of 477 and 575 nucleotides. The precursors of proline-rich E. andrei lumbricin and the related peptide contain 63 and 59 amino acids, respectively. Phylogenetic analysis indicated close relationship with other annelid lumbricins. Highest expression of both mRNAs appeared in the proximal part of the intestine (pharynx, gizzard), while other tested organs had moderate (body wall, midgut, ovary, metanephridium, seminal vesicles, ventral nerve cord) or low (coelomocytes) levels. During ontogenesis their expression revealed continuous increase in embryos. Following 48 h of in vivo Gram-positive bacteria challenge both mRNAs were significantly elevated in coelomocytes, while Gram-negative bacteria or zymosan stimulation had no detectable effects.


Assuntos
Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Positivas/imunologia , Proteínas de Helminto/genética , Intestinos/fisiologia , Oligoquetos/imunologia , Peptídeos/genética , Animais , Anti-Infecciosos/metabolismo , Clonagem Molecular , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Helminto/metabolismo , Imunidade Inata , Oligoquetos/genética , Oligoquetos/microbiologia , Peptídeos/metabolismo , Filogenia , Transcriptoma
15.
Nanoscale ; 10(46): 21960-21970, 2018 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-30444228

RESUMO

Titanium dioxide (TiO2) based nanomaterials (NMs) are among the most produced NMs worldwide. When irradiated with light, particularly UV, TiO2 is photoactive, a property that is explored for several purposes. There are an increasing number of reports on the negative effects of photoactivated TiO2 on non-target organisms. We have here studied the effect of a suite of reference type TiO2 NMs (i.e. NM103, NM104, and NM105 and compared these to the bulk) with and without UV radiation to the oligochaete Enchytraeus crypticus. High-throughput gene expression was used to assess the molecular mechanisms, while also anchoring it to the known effects at the organism level (i.e., reproduction). Results showed that the photoactivity of TiO2 (UV exposed) played a major role in enhancing TiO2 toxicity, activating the transcription of oxidative stress, lysosome damage and apoptosis mechanisms. For non-UV activated TiO2, where toxicity at the organism level (reproduction) was lower, results showed potential for long-term effects (i.e., mutagenic and epigenetic). NM specific mechanisms were identified: NM103 affected transcription and translation, NM104_UV negatively affected the reproductive system/organs, and NM105_UV activated superoxide anion response. Results provided mechanistic information on UV-related phototoxicity of TiO2 materials and evidence for the potential long-term effects.


Assuntos
Nanopartículas Metálicas/química , Oligoquetos/genética , Titânio/química , Animais , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/efeitos da radiação , Nanopartículas Metálicas/toxicidade , Oligoquetos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/efeitos da radiação , Superóxidos/metabolismo , Raios Ultravioleta
16.
J Agric Food Chem ; 66(45): 11902-11908, 2018 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-30372061

RESUMO

Testing for effects of pesticides on nontarget organisms is an integral part of ecological risk assessment. In the present study, the acute toxicity of sulfoxaflor to earthworms was evaluated using an artificial soil toxicity test, and sublethal effects were assessed through oxidative stress and metabolomics. Sulfoxaflor is a supertoxic pollutant to earthworms that easily bioaccumulates in earthworms, and contains LC2, LC10, and LC50 values of 0.08 (0.04-0.13), 0.19 (0.11-0.25), and 0.54 (0.45-0.65) mg/kg, respectively. Sublethal doses of sulfoxaflor resulted in oxidative damage to earthworms in which antioxidant enzymatic activities including SOD, CAT, and GST were significantly inhibited and MDA content accumulated. Metabolomics analysis suggested that the energy metabolism and the urea cycle in earthworms were significantly activated, while nucleotide metabolism was depressed, which could cause DNA damage. The results suggest earthworms have the potential to be a new entry point for sulfoxaflor into the wildlife food chain. Since earthworms significantly contribute to soil function and ecosystems, the high safety risks of sulfoxaflor to the earthworm could extend to the environment. In view of these findings, more attention should be given to the risks sulfoxaflor poses to the environment through its effects on earthworms.


Assuntos
Oligoquetos/efeitos dos fármacos , Praguicidas/toxicidade , Piridinas/toxicidade , Poluentes do Solo/toxicidade , Compostos de Enxofre/toxicidade , Animais , Catalase/genética , Catalase/metabolismo , Dano ao DNA/efeitos dos fármacos , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Malondialdeído/metabolismo , Oligoquetos/genética , Oligoquetos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo
17.
PLoS One ; 13(9): e0204234, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30260966

RESUMO

Earthworms show a wide spectrum of regenerative potential with certain species like Eisenia fetida capable of regenerating more than two-thirds of their body while other closely related species, such as Paranais litoralis seem to have lost this ability. Earthworms belong to the phylum Annelida, in which the genomes of the marine oligochaete Capitella telata and the freshwater leech Helobdella robusta have been sequenced and studied. Herein, we report the transcriptomic changes in Eisenia fetida (Indian isolate) during regeneration. Following injury, E. fetida regenerates the posterior segments in a time spanning several weeks. We analyzed gene expression changes both in the newly regenerating cells and in the adjacent tissue, at early (15days post amputation), intermediate (20days post amputation) and late (30 days post amputation) by RNAseq based de novo assembly and comparison of transcriptomes. We also generated a draft genome sequence of this terrestrial red worm using short reads and mate-pair reads. An in-depth analysis of the miRNome of the worm showed that many miRNA gene families have undergone extensive duplications. Sox4, a master regulator of TGF-beta mediated epithelial-mesenchymal transition was induced in the newly regenerated tissue. Genes for several proteins such as sialidases and neurotrophins were identified amongst the differentially expressed transcripts. The regeneration of the ventral nerve cord was also accompanied by the induction of nerve growth factor and neurofilament genes. We identified 315 novel differentially expressed transcripts in the transcriptome, that have no homolog in any other species. Surprisingly, 82% of these novel differentially expressed transcripts showed poor potential for coding proteins, suggesting that novel ncRNAs may play a critical role in regeneration of earthworm.


Assuntos
Perfilação da Expressão Gênica/métodos , Redes Reguladoras de Genes , Oligoquetos/fisiologia , Análise de Sequência de DNA/métodos , Animais , Evolução Molecular , Regulação da Expressão Gênica , Genoma , MicroRNAs/genética , Família Multigênica , Oligoquetos/genética , Filogenia , Regeneração , Fatores de Transcrição SOXC/genética , Análise de Sequência de RNA/métodos
18.
PLoS One ; 13(9): e0204469, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30240427

RESUMO

Uniformly pigmented Eisenia andrei (Ea) and striped E. fetida (Ef) lumbricid earthworms are hermaphrodites capable of self-fertilization, cross-fertilization, and asymmetrical hybridization. The latter was detected by genotyping of F1 and F2 progeny of the controlled Ea+Ef pairs by species-specific sequences of maternal mitochondrial COI genes and maternal/paternal nuclear S28 rRNA genes. Among F1offspring there were self-fertilized Ea (aAA), Ef (fFF), and cross-fertilized fertile Ea-derived hybrids (aAF); the latter mated with Ea and gave new generation of Ea and hybrids, while mated with Ef gave Ea, Ef, Ea-derived hybrids and sterile Ef-derived hybrids (fFA). Coelomic fluid of Ea exhibits unique fluorescence spectra called here the M-fluorescence considered as a molecular biomarker of this species. Since similar fluorescence was detected also in some Ef (hypothetical hybrids?), the aim of present investigations was to identify the M-positive earthworms among families genotyped previously. It was assumed that factor/s responsible for metabolic pathways leading to production of undefined yet M-fluorophore might be encoded/controlled by alleles of hypothetical nuclear gene of Eisenia sp. segregating independently from species-specific S28 rRNA nuclear genes, where 'MM' or 'Mm' alleles determine M-positivity while 'mm' alleles determine M-negative phenotypes. Spectra of M-fluorescence were detected in all 10 Ea (aAAMM) and 19 Ea-derived hybrids (aAFMm), three of four Ef-derived hybrids (fFAMm) and one 'atypical' Ef (fFFMm) among 13 Ef earthworms. Among progeny of 'atypical' M-positive Ef (fFFMm) reappeared 'typical' M-negative Ef (fFFmm), confirming such hypothesis. Alternatively, the M-fluorescence might be dependent on unknown gene products of vertically-transmitted Ea-specific symbiotic bacteria sexually transferred to the Ef partner. Hypotheses of intrinsic and external origin of M-fluorescence might complement each other. The presence/absence of M-fluorophore does not correspond with body pigmentation patterns; Ef-characteristic banding appeared in posterior parts of hybrids body. In conclusion, Ea/Ef hybridization may serve for further studies on bi-directional gene flow.


Assuntos
Fluxo Gênico , Hibridização Genética , Oligoquetos/genética , Alelos , Animais , Fluorescência , Genótipo , Oligoquetos/metabolismo , Fenótipo , Pigmentação/genética , RNA Ribossômico 28S , Especificidade da Espécie
19.
J Aquat Anim Health ; 30(4): 264-279, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30133011

RESUMO

Placer Creek, a tributary of Sangre de Cristo Creek in Colorado's San Luis Valley, supported an allopatric core conservation population of native Rio Grande Cutthroat Trout Oncorhynchus clarkii virginalis during much of the 20th century. After the failure of gabion barriers in the late 1990s, Brook Trout Salvelinus fontinalis infected with Myxobolus cerebralis invaded from Sangre de Cristo Creek. By 2005, whirling disease (WD) and competition from Brook Trout reduced Rio Grande Cutthroat Trout numbers to less than 10% of the total trout population. New barriers were constructed in 2006 and the stream was treated with rotenone in 2007 and 2009 to eliminate all fish prior to the reintroduction of Rio Grande Cutthroat Trout. Results of WD research studies in Montana, California, and Colorado indicated it might be possible to break the life cycle of the parasite in some situations. Our management interventions included (1) reducing the fish population in the stream to zero for approximately 14 months, (2) introducing lineage V and VI Tubifex tubifex worms, which are not susceptible to M. cerebralis, and (3) eliminating a small off-channel pond that provided optimal habitat that sustained a localized high-density population of lineage III T. tubifex, the oligochaete host susceptible to M. cerebralis. Electrofishing during the fall of 2009 and spring of 2010 indicated the drainage was devoid of fish. Fry, juvenile, and adult Rio Grande Cutthroat Trout were stocked in September and October of 2010 and 2011. Approximately 975,000 lineage V and VI T. tubifex were introduced into Placer Creek between 2010 and 2012 as possible oligochaete competitors for the lineage III worms. The off-channel pond was filled in, and the surface was reseeded in April 2012. No evidence of M. cerebralis infection was detected among more than 280 Rio Grande Cutthroat Trout tested between July 2012 and July 2016, indicating the parasite had been eradicated from the Placer Creek basin upstream of the barriers.


Assuntos
Doenças dos Peixes/parasitologia , Myxobolus , Oncorhynchus/parasitologia , Animais , Colorado , Reservatórios de Doenças/parasitologia , Doenças dos Peixes/prevenção & controle , Oligoquetos/genética , Oligoquetos/parasitologia , Doenças Parasitárias em Animais/parasitologia , Doenças Parasitárias em Animais/prevenção & controle , Rios , Rotenona , Truta/parasitologia
20.
Chemosphere ; 212: 87-94, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30142569

RESUMO

Gene expression can vary with the organisms' life stage. It is known that embryos can be more sensitive to toxicant exposure, as previously demonstrated for Enchytraeus crypticus (Oligochaeta) exposed to cadmium (Cd), known to cause embryotoxicity and hatching delay. It was shown that Ca enters embryos via the L-type Ca channels in the cocoon membrane, this being affected in Cd exposed embryos (Cd-Ca competition is well-known). In the present study, the embryotoxic mechanisms of Cd were studied via high-throughput gene expression for E. crypticus. Cocoons (1-2 days old), instead of the adult organism, were exposed in Cd spiked LUFA 2.2 soil during 1 day. Results showed that Cd affected Ca homeostasis which is implicated in several other molecular processes. Several of the major modulators of Cd toxicity (e.g., impaired gene expression, cell cycle arrest, DNA and mitochondrial damage) were identified in the embryos showing its relevancy as a model in ecotoxicogenomics. The draft Adverse Outcome Pathway was improved. Previously was hypothesized that gene regulation mechanisms were activated to synthesize more Ca channel proteins - this was confirmed here. Further, novel evidences were that, besides the extracellular competition, Cd competes intracellularly which causes a reduction in Ca efflux, and potentiates Cd embryotoxicity.


Assuntos
Cádmio/toxicidade , Expressão Gênica , Oligoquetos/embriologia , Animais , Cálcio/metabolismo , Invertebrados/embriologia , Invertebrados/genética , Oligoquetos/genética , Oligoquetos/fisiologia , Solo/química , Poluentes do Solo/toxicidade
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