Abstract
Vermicomposting is a process of degradation of biowaste which involves complex interactions between earthworms and microorganisms. This process lacks a thermophilic stage and thus, the possible presence of pathogens poses a potential health hazard. To assess the contribution of earthworms during the selective reduction of various pathogens, apple pomace substrate was artificially inoculated with Escherichia coli, Salmonella spp., thermotolerant coliform bacteria, and Enterococci. The artificial bacterial load did not influence the weight, reproduction, or intestinal enzymatic activity of the earthworms, but it caused reversible histological changes to the epithelial layer and chloragogen tissue of their intestines. The reduction of pathogenic Enterococci and E. coli from the substrate was accelerated by earthworms (63-fold, 77-fold, and 840-fold for Enterococci and 6-fold, 36-fold, and 7-fold for E. coli inoculated substrates after 2, 4, and 6 weeks, respectively). Moreover, the rapid elimination of Salmonella spp. was supported by the upregulated expression of two pattern recognition receptors which bind lipopolysaccharide, coelomic cytolytic factor, and lipopolysaccharide-binding protein. Further, the microbiomes of the intestine and the composting substrate differed significantly.
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- CCF:
-
Coelomic cytolytic factor
- CFU:
-
Colony-forming unit
- HTS:
-
High-throughput sequencing
- PCR:
-
Polymerase chain reaction
- PRRs:
-
Pattern recognition receptors
- TCB:
-
Thermotolerant coliform bacteria
- TLR:
-
Toll-like receptor
References
Aira M, Gomez-Brandon M, Gonzalez-Porto P, Dominguez J (2011) Selective reduction of the pathogenic load of cow manure in an industrial-scale continuous-feeding vermireactor. Bioresour Technol 102:9633–9637. https://doi.org/10.1016/j.biortech.2011.07.115
Aira M, Bybee S, Perez-Losada M, Dominguez J (2015) Feeding on microbiomes: effects of detritivory on the taxonomic and phylogenetic bacterial composition of animal manures. FEMS Microbiol Ecol 91
Akira S, Uematsu S, Takeuchi O (2006) Pathogen recognition and innate immunity. Cell 124:783–801. https://doi.org/10.1016/j.cell.2006.02.015
Aronesty E (2013) Comparison of sequencing utility programs. Open Bioinf J 7:1–8
Bilej M, Rossmann P, Sinkora M, Hanusova R, Beschin A, Raes G, De Baetselier P (1998) Cellular expression of the cytolytic factor in earthworms Eisenia foetida. Immunol Lett 60:23–29
Castillo JM, Romero E, Nogales R (2013) Dynamics of microbial communities related to biochemical parameters during vermicomposting and maturation of agroindustrial lignocellulose wastes. Bioresour Technol 146:345–354. https://doi.org/10.1016/j.biortech.2013.07.093
Cole JR, Wang Q, Fish JA, Chai B, McGarrell DM, Sun Y, Brown CT, Porras-Alfaro A, Kuske CR, Tiedje JM (2014) Ribosomal Database Project: data and tools for high throughput rRNA analysis. Nucleic Acids Res 42:D633–D642
Dominguez J and Gomez-Brandon M (2012) Vermicomposting: composting with earthworms to recycle organic wastes. In: Kumar S, Bharti A (eds) Management of Organic Waste. Intech Open Science. https://doi.org/10.5772/33874
Drake HL, Horn MA (2007) As the worm turns: the earthworm gut as a transient habitat for soil microbial biomes. Annu Rev Microbiol 61:169–189. https://doi.org/10.1146/annurev.micro.61.080706.093139
Dvorak J, Mancikova V, Pizl V, Elhottova D, Silerova M, Roubalova R, Skanta F, Prochazkova P, Bilej M (2013) Microbial environment affects innate immunity in two closely related earthworm species Eisenia andrei and Eisenia fetida. PLoS One 8:e79257. https://doi.org/10.1371/journal.pone.0079257
Dvorak J, Roubalova R, Prochazkova P, Rossmann P, Skanta F, Bilej M (2016) Sensing microorganisms in the gut triggers the immune response in Eisenia andrei earthworms. Dev Comp Immunol 57:67–74
Edgar RC (2013) UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nat Methods 10:996–998. https://doi.org/10.1038/Nmeth.2604
Edwards CA (2011) Human pathogen reduction during vermicomposting. In: Edwards CA, Arancon NQ, Sherman R (eds) Vermiculture Technology: Earthworms, Organic Wastes and Environmental Management. CRC Press
Finola M, Rodriguez C, Beoletto V (1995) Bacteriology of the gastrointestinal tract of earthworm Eisenia fetida (Annelida, Oligochaeta) cultivated in composted broiler litter. Rev Argent Microbiol 27:210–213
Godfree A, Farrell J (2005) Processes for managing pathogens. J Environ Qual 34:105–113
Graves A, Weaver RW, Entry J (2009) Characterization of enterococci populations in livestock manure using BIOLOG. Microbiol Res 164:260–266
Hammer O (2011) Data analysis with PAST for micropaleontologists. Grzyb Found Spec Pub 17:90–90
Hanc A, Chadimova Z (2014) Nutrient recovery from apple pomace waste by vermicomposting technology. Bioresour Technol 168:240–244. https://doi.org/10.1016/j.biortech.2014.02.031
Hanc A, Pliva P (2013) Vermicomposting technology as a tool for nutrient recovery from kitchen bio-waste. J Mat Cycl and Waste Manag 15:431–439
Huang K, Xia H, Cui GY, Li FS (2017) Effects of earthworms on nitrification and ammonia oxidizers in vermicomposting systems for recycling of fruit and vegetable wastes. Sci Total Environ 578:337–345. https://doi.org/10.1016/j.scitotenv.2016.10.172
Joskova R, Silerova M, Prochazkova P, Bilej M (2009) Identification and cloning of an invertebrate-type lysozyme from Eisenia andrei. Dev Comp Immunol 33:932–938. https://doi.org/10.1016/j.dci.2009.03.002
Kiernan JA (2008) Nuclear stains. CSH Protoc 2008:pdb top50. https://doi.org/10.1101/pdb.top50
Malinska K, Zabochnicka-Swiatek M, Caceres R, Marfa O (2016) The effect of precomposted sewage sludge mixture amended with biochar on the growth and reproduction of Eisenia fetida during laboratory vermicomposting. Ecol Eng 90:35–41. https://doi.org/10.1016/j.ecoleng.2016.01.042
Milochau A, Lassegues M, Valembois P (1997) Purification, characterization and activities of two hemolytic and antibacterial proteins from coelomic fluid of the annelid Eisenia fetida andrei. Biochim Biophys Acta 1337:123–132
Monroy F, Aira M, Dominguez J (2009) Reduction of total coliform numbers during vermicomposting is caused by short-term direct effects of earthworms on microorganisms and depends on the dose of application of pig slurry. Sci Total Environ 407:5411–5416. https://doi.org/10.1016/j.scitotenv.2009.06.048
Pachepsky YA, Sadeghi AM, Bradford SA, Shelton DR, Guber AK, Dao T (2006) Transport and fate of manure-borne pathogens: modeling perspective. Agric Water Manag 86:81–92
Pass DA, Morgan AJ, Read DS, Field D, Weightman AJ, Kille P (2015) The effect of anthropogenic arsenic contamination on the earthworm microbiome. Environ Microbiol 17:1884–1896. https://doi.org/10.1111/1462-2920.12712
Prochazkova P, Silerova M, Felsberg J, Joskova R, Beschin A, De Baetselier P, Bilej M (2006) Relationship between hemolytic molecules in Eisenia fetida earthworms. Dev Comp Immunol 30:381–392
Roubalova R, Dvorak J, Prochazkova P, Elhottova D, Rossmann P, Skanta F, Bilej M (2014) The effect of dibenzo-p-dioxin- and dibenzofuran-contaminated soil on the earthworm Eisenia andrei. Environ Pollut 193:22–28
Sekizawa Y, Kubo T, Kobayashi H, Nakajima T, Natori S (1997) Molecular cloning of cDNA for lysenin, a novel protein in the earthworm Eisenia foetida that causes contraction of rat vascular smooth muscle. Gene 191:97–102
Skanta F, Roubalova R, Dvorak J, Prochazkova P, Bilej M (2013) Molecular cloning and expression of TLR in the Eisenia andrei earthworm. Dev Comp Immunol 41:694–702. https://doi.org/10.1016/j.dci.2013.08.009
Skanta F, Prochazkova P, Roubalova R, Dvorak J, Bilej M (2016) LBP/BPI homologue in Eisenia andrei earthworms. Dev Comp Immunol 54:1–6. https://doi.org/10.1016/j.dci.2015.08.008
Soobhany N, Mohee R, Garg VK (2017) Inactivation of bacterial pathogenic load in compost against vermicompost of organic solid waste aiming to achieve sanitation goals: a review. Waste Manag 64:51–62. https://doi.org/10.1016/j.wasman.2017.03.003
Swati A, Hait S (2018) A comprehensive review of the fate of pathogens during vermicomposting of organic wastes. J Environ Qual 47:16–29. https://doi.org/10.2134/jeq2017.07.0265
Titilawo Y, Obi L, Okoh A (2015) Occurrence of virulence gene signatures associated with diarrhoeagenic and non-diarrhoeagenic pathovars of Escherichia coli isolates from some selected rivers in south-western Nigeria. BMC Microbiol 15:204. https://doi.org/10.1186/s12866-015-0540-3
Turner C (2002) The thermal inactivation of E-coli in straw and pig manure. Bioresour Technol 84:57–61
Vetrovsky T, Baldrian P (2013) Analysis of soil fungal communities by amplicon pyrosequencing: current approaches to data analysis and the introduction of the pipeline SEED. Biol Fert Soils 49:1027–1037. https://doi.org/10.1007/s00374-013-0801-y
Yu Y, Lee C, Kim J, Hwang S (2005) Group-specific primer and probe sets to detect methanogenic communities using quantitative real-time polymerase chain reaction. Biotechnol Bioeng 89:670–679. https://doi.org/10.1002/bit.20347
Acknowledgements
We thank L. Matějů for technical help with selective bacterial cultivation.
Funding
This research was supported by the Institutional Research Concept RVO 61388971 and by the Ministry of Agriculture of the Czech Republic under the NAZV project No. QJ1530034 and by CULS Prague under the CIGA project No. 20172018.
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Responsible editor: Diane Purchase
Highlights
• Overdosed bacterial load did not influence earthworms’ weight or reproduction
• Bacterial load caused reversible histological changes of earthworms’ intestine
• Reduction of E. coli and Enterococci pathogens was accelerated by earthworms
• Overdose of Salmonella spp. elicited higher mRNA levels of molecules binding LPS
• Intestine microbiome of earthworms differed from microbiome of substrate
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Procházková, P., Hanč, A., Dvořák, J. et al. Contribution of Eisenia andrei earthworms in pathogen reduction during vermicomposting. Environ Sci Pollut Res 25, 26267–26278 (2018). https://doi.org/10.1007/s11356-018-2662-2
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DOI: https://doi.org/10.1007/s11356-018-2662-2