• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.885-892
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• 2011

The bacterial communities in the intestinal tracts of earthworm were investigated by culture-dependent and -independent approaches. In total, 72 and 55 pure cultures were isolated from the intestinal tracts of earthworms under aerobic and anaerobic conditions, respectively. Aerobic bacteria were classified as Aeromonas (40%), Bacillus (37%), Photobacterium (10%), Pseudomonas (7%), and Shewanella (6%). Anaerobic bacteria were classified as Aeromonas (52%), Bacillus (27%), Shewanella (12%), Paenibacillus (5%), Clostridium (2%), and Cellulosimicrobium (2%). The dominant microorganisms were Aeromonas and Bacillus species under both aerobic and anaerobic conditions. In all, 39 DNA fragments were identified by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis. Aeromonas sp. was the dominant microorganism in feeds, intestinal tracts, and casts of earthworms. The DGGE band intensity of Aeromonas from feeds, intestinal tracts, and casts of earthworms was 12.8%, 14.7%, and 15.1%, respectively. The other strains identified were Bacillus, Clostridium, Enterobacter, Photobacterium, Pseudomonas, Shewanella, Streptomyces, uncultured Chloroflexi bacterium, and uncultured bacterium. These results suggest that PCR-DGGE analysis was more efficient than the culturedependent approach for the investigation of bacterial diversity and the identification of unculturable microorganisms.

• The Korean Journal of Mycology
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• v.50 no.4
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• pp.373-381
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• 2022

Strain E4, an unrecorded species of dimorphic fungi, was isolated from the gut of earthworms collected in Gyeonggi Province, South Korea. Nucleotide sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region revealed that this species is a member of the genus Blastobotrys, Blastobotrys illinoisensis. Strain E4 differed from its closest known species, B. mokoenaii and B. malaysiensis, by harboring 3-5 and 12-14 nucleotide substitutions in the D1/D2 and ITS regions, respectively. Phylogenetic analysis based on concatenated sequences of the D1/D2 region of the LSU rRNA gene and the ITS region also indicated that strain E4 belongs to the Blastobotrys clade and is distinct from other related species in the clade. The previously unreported isolate could be distinguished from closely related species by its inability to ferment carbon sources. To our knowledge, this is the first report on the isolation of Blastobotrys species from the gut of earthworms in Korea. The strain used was E4 (=KCTC 27831=JCM 33428).

• International Journal of Industrial Entomology and Biomaterials
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• v.48 no.2
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• pp.67-77
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• 2024

This study aims to assess the efficacies of exogenous enzymes, derived from invertebrate gut-associated microbes, as feed additives, in reducing volatile organic compound (VOC) emissions using an in vitro pig intestine continuous fermentation system. An in vitro continuous fermentation model was used to simulate a comparable bionic digestion system by co-reacting feed, enzymatic additives (arazyme, mannanase, and xylanase, derived from the gut bacteria of Nephila clavata, Eisenia fetida, and Moechotypa diphysis, respectively), and gastrointestinal microbes, followed by an analysis of their correlations. A significant correlation was observed between exogenous enzyme supplementation and reduced VOC emissions in the fecal phase of continuous fermentation (p < 0.05). The concentration of VOCs decreased by 3.75 and 2.75 ppm in the treatment group following arazyme and multi-enzyme supplementation, respectively, compared to that in the control group (7.83 ppm). In addition, supplementation with arazyme and multiple enzymes significantly affected the microbial composition of each fermentation phase (p < 0.05). In particular, Lactiplantibacillus pentosus and Pediococcus pentosaceus, which changed in abundance according to arazyme or multi-enzyme supplementation, exhibited a positive relationship with VOC emissions. These results suggest that exogenous enzymes derived from invertebrate gut-associated bacteria can be efficiently applied as feed additives, leading to a reduction in VOC emissions.

• Korean Journal of Environmental Biology
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• v.28 no.4
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• pp.212-217
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• 2010

Soil pollution by heavy metals has become a significant environmental concern due to a variety of human activities. Specially toxicity caused by excessive mercury exposure is now being recognized as a widespread environmental problem and is continuing to attract a great deal of public concerns. The earthworms are very important animals that aerate the soil with their burrowing action and enrich the soil by decomposing organic matters. Especially the earthworm Eisenia fetida is routinely used in ecotoxicological studies. The levels of DNA damage in earthworms treated with HgCl2 and ionizing radiation were investigated in this study. Genotoxic effects were evaluated in the earthworm's coelomocytes using the comet assay (Single Cell Gel Electrophoresis; SCGE). The results showed that the mercury chloride and radiation were responsible for the genotoxic effects on earthworms. The level of DNA damage significantly increased after the treatment of mercury chloride combined with ionizing radiation. The combined treatment of $HgCl_2$ and ionizing radiation had a greater genotoxicity. This study is amenable to further study such as enzyme activation assay.

• Korean Journal of Environmental Biology
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• v.27 no.1
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• pp.40-47
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• 2009

Heavy reliance on organosynthetic chemical methods to control agricultural pest tends to deteriorate and lower fertility of soil. We studied annual occurrence of earthworm in environment-friendly agroecosystem and attempted to determine the degree of soil contamination. Candidate bioindicators of earthworm in agroecosystem were selected through analysis of earthworm diversities. Depending on the degree of pesticide use, the fields were devided into conventional, less-pesticide, pesticide free, and organic fields. A nearby field not in agricultural use was chosen as a control. This study was carried out in the greenhouses (lettuce and grape) in 2006 and in orchards (apple, pear, peach, citrus and grape) in 2007. Earthworm was collected from 50 $\times$ 50 $\times$ 10 cm$^3$ of respective soil. We found 11 species from 5 genera and 3 families, and 12 species from 4 genera and 3 families in the greenhouses and in orchards, respectively. The contents of organic matter in soil in the greenhouses were variable but quite less so in orchards. Amynthas heteropodus occurred most frequently in both sites suggesting that it could be adopted as a bioindicator in agroecosystem in Korea.

• Journal of the Korea Organic Resources Recycling Association
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• v.16 no.4
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• pp.64-73
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• 2008

The cDNA of CCF (coelomic cytolytic factor)-like gene (EC 3.2.1.16), a kind of glycosyl hydorlase, was isolated and cloned from the midgut of the earthworm Eisenia anderi. The size of nucleotide sequence appeared to be 1,152 bp and its predicted coding region was composed of 384 amino acid residues including the initiation methionine. The 17 residues at N-terminal end in the deduced amino acid sequence were regarded to be a signal peptide. Based on the amino acid sequence analysis, it appeared that this CCF-like protein could belong to glycosyl hydrolase family 16 (GHF16) and showed a high sequence homology of about 79~99% with CCF and CCF-like proteins from other earthworm species. The CCFs and CCF-like proteins from various earthworm species exhibited a 100% homology in the polysacchride-binding motif and glucanase motif. It has been reported that the CCFs isolated from E. fedita appeared to show a broader pattern recognition specificity than those from other earthworm species because this species resides in decaying organic matter showing very high microbial activity, implying that CCF-like protein isolated in this study from E. andrei might exhibit a broad substrate specificity that is a useful characteristic for industrial application. A phylogenetic analysis using the deduced amino acid sequences of CCF-related proteins through the BLASTX revealed that GHF16 families could be divided into three groups of metazoa, viriplantae and eubacteria subfamily. Subsequently the CCF-related proteins of metazoa subfamily could clearly be subgroup into lophotrochozoan and edysozoan type including a deuterostome origin. Further understanding of the biological properties of E. andrei CCF-like protein should be addressed to regulate the ${\beta}$-D-glucan hydrolysis and production for the industrial uses.

• Journal of Soil and Groundwater Environment
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• v.19 no.5
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• pp.35-44
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• 2014

In this study, the bio-phytoremediation and phytoremediation technologies were applied to the soils contaminated with total petroleum hydrocarbons (TPH) and heavy metals to evaluate the remediation efficacy from May 2012 to December 2013. Poplar (Populus bonatii Levl.) and Sun Hemp (Crotalaria juncea L.) were selected and planted in phytoremediation practice. These plants were also utilized in the bio-phytoremediation practice, with the addition of earthworm (Eisenia fetida) and petroleum-degrading bacteria (Pseudomonos sp. NKNU01). Furthermore, physiological characteristics, such as photosynthesis rate and maximal photochemical yield, of all testing plants were also measured in order to assess their health conditions and tolerance levels in adverse environment. After 20 months of remedial practice, the results showed that bio-phytoremediation practice had a higher rate of TPH removal efficacy at 30-60 cm depth soil than that of phytoremediation. However, inconsistent results were discovered while analyzing the soil at 100 cm depth. The study also showed that the removal efficiency of heavy metals was lower than that of TPH after remediation treatment. The results from test field tissue sample analysis revealed that more Zinc than Chromium was absorbed and accumulated by the tested plants. Plant height measurements of Poplar and Sun Hemp showed that there were insignificant differences of growth between the plants in remediation plots and those in the control plot. Physiological data of Poplar also suggested it has higher tolerance level toward the contaminated soils. These results indicated that the two testing plants were healthy and suitable for this remediation study.

• Environmental Analysis Health and Toxicology
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• v.20 no.4 s.51
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• pp.279-286
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• 2005

Earthworm (Eisenia fetida) acute toxicity test was carried out and ecological risk assessment in soil was performed with national monitoring data. 14 day - $LC_{50}$ of nonylphenol and bisphenol A were 288.1 mg/kg and 90.1 mg/kg, respectively. And NOECs of nonylphenol and bisphenol A were 250 mg/kg and 50 mg/kg, respectively. Significant weight decrement was appeared at 70 mg/kg of bisphenol A, however, nonylphenol at concentrations tested did not severe adverse effect on the weight decrement. The environmental monitoring has been carrying out by NIER since 1999. Exposure levels of nonylphenol in soil were ND$\sim$10.55 $\mu$g/kg and those of bisphenol A were ND$\sim$15.50$\mu$g/kg in National Monitoring data which had been performed from 2000 to 2004. The measured soil exposure level was applied to evaluate the environmental risk assessment. The values of PNEC for bisphenol A and nonylphenol were determined as 0.5 mg/kg and 2.5 mg/kg, respectively using the safety factors which were suggested in EU and OECD. The values of HQ (PEC/PNEC) were determined to be below I for bisphenol A and nonylphenol when the maximum exposure levels for bispheol A (15.50$\mu$g/kg) and nonylphenol (10.55$\mu$g/kg) were applied. Conclusively, the environmental risk assessment of bisphenol A and nonylphenol was not critical in soil.

• Journal of Applied Biological Chemistry
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• v.50 no.4
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• pp.281-287
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• 2007

Microorganisms were isolated from earthworm intestine and examined for their ability to degrade 3-methyl-4-nitrophenol (MNP), a main degradation product of the insecticide fenitrothion. An isolate that showed the best degradation of MNP was selected for further study. The 16S rRNA analysis showed that the isolate belongs to the genus of Burkholderia, close to phenanthrene-degrading Burkholderia sp. S4.9, and is named Burkholderia sp. SH-1. When time-course degradation of MNP by SH-1 was examined by high performance liquid chromatographic analysis, almost complete degradation of MNP was observed within 26 h. Colony forming unit value assays indicated that the isolate SH-1 was capable of utilizing MNP as a sole carbon source. SH-1 could also degrade p-nitrophenol (PNP) but could not degrade ortho-substituted nitroaromatics such as 2,4-, 2,6- and 2,5-dinitrophenol. Catechol was detected as the main degration product of MNP and PNP. SH-1 was also found in the soil from which earthworms were obtained. These results suggest that the dispersal of Burkholderia sp. SH-1 into different environment with the aid of earthworms is likely to play a role in bioremediation of the soil contaminated with MNP.

• Korean Journal of Soil Science and Fertilizer
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• v.40 no.1
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• pp.51-56
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• 2007

The toxicities of contaminated soils with 8 consecutive year applications of three levels (12.5, 25.0, and $50.0t\;dry\;matter\;ha^{-1}yr^{-1}$) of four organic sludge [municipal sewage sludge (MSS), industrial sewage sludge (ISS), alcohol fermentation processing sludge (AFPS) and leather processing sludge (LPS)] on earthworm (Eisenia fetida) were examined by using microcosm container in the laboratory. Results were compared with those of pig manure compost (PMC) treated soil. In tests with three treatment levels (12.5, 25.0, and 50.0 t per plot), ISS treated soil showed higher contents of Cu (18.9~26.2 fold), Cr (7.7~34.7 fold), and Ni (14.8~18.8 fold) at 8 years post treatment, than PMC treated soil. LPS treated soil showed higher contents of Cr (35.7~268.0 fold) and Ni (4.5~7.6 fold) than PMC treated soil. There were no great differences in heavy metal contents among MSS, AFPS, and PMC treated soils. In these contaminated soils, earthworm mortalities of MSS and AFPS treated soils at 8 weeks post-exposure were similar to those of PMC treated soil regardless of each treatment level. Toxic effect (26.7~96.7 mortality) on the ISS and LPS treated soils was significantly higher than one of PMC treated soil, with an exception of LPS soil treated with 25.0 t per plot. At 16 weeks post-exposure, earthworm mortalities of AFPS' 12.5 and 25.0 t treated soils were similar to those of PMC treated soil. Toxic effect (53.3~100 mortality) on the 12.5, 25.0, and 50.0 t treated soils of MSS, ISS and LPS, and AFPS' 50.0 t treated soils was significantly higher than those of PMC treated soil. The data suggested that the 12.5, 25.0, and 50.0 t of MSS, ISS and LPS, and AFPS' 50.0 t treated soils were evaluated to have toxicity on earthworm.