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Dechloromonas denitrificans sp. nov., Flavobacterium denitrificans sp. nov., Paenibacillus anaericanus sp. nov. and Paenibacillus terrae strain MH72, N₂O-producing bacteria isolated from the gut of the earthworm Aporrectodea caliginosa

Marcus A. Horn¹, Julian Ihssen¹, Carola Matthies^1,, Andreas Schramm^1,, Georg Acker² and Harold L. Drake¹

¹ Department of Ecological Microbiology, University of Bayreuth, 95440 Bayreuth, Germany
² Electron Microscopy Laboratory, University of Bayreuth, 95440 Bayreuth, Germany

Correspondence
Harold L. Drake
hld{at}uni-bayreuth.de

Earthworms emit nitrous oxide (N₂O) via the activity of bacteria in their gut. Four N₂O-producing facultative aerobes, ED1^T, ED5^T, MH21^T and MH72, were isolated from the gut of the earthworm Aporrectodea caliginosa. The isolates produced N₂O under conditions that simulated the microenvironment of the earthworm gut. ED1^T and ED5^T were Gram-negative, motile rods that carried out complete denitrification (i.e. the reduction of nitrate to N₂) and contained membranous c-type cytochromes. ED1^T grew optimally at 30 °C and pH 7. ED1^T oxidized organic acids and reduced (per)chlorate, sulfate, nitrate and nitrite. The closest phylogenetic relative of ED1^T was Dechloromonas agitata. ED5^T grew optimally at 25 °C and pH 7. ED5^T grew mainly on sugars, and nitrate and nitrite were used as alternative electron acceptors. The closest phylogenetic relatives of ED5^T were Flavobacterium johnsoniae and Flavobacterium flevense. MH21^T and MH72 were motile, spore-forming, rod-shaped bacteria with a three-layered cell wall. Sugars supported the growth of MH21^T and MH72. Cells of MH21^T grew in chains, were linked by connecting filaments and contained membranous b-type cytochromes. MH21^T grew optimally at 30–35 °C and pH 7·7, grew by fermentation and reduced low amounts of nitrite to N₂O. The closest phylogenetic relatives of MH21^T were Paenibacillus borealis and Paenibacillus chibensis. Based on morphological, physiological and phylogenetic characteristics, ED1^T (=DSM 15892^T=ATCC BAA-841^T), ED5^T (=DSM 15936^T=ATCC BAA-842^T) and MH21^T (=DSM 15890^T=ATCC BAA-844^T) are proposed as type strains of the novel species Dechloromonas denitrificans sp. nov., Flavobacterium denitrificans sp. nov. and Paenibacillus anaericanus sp. nov., respectively. MH72 is considered a new strain of Paenibacillus terrae.

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of Dechloromonas denitrificans ED1^T, Flavobacterium denitrificans ED5^T and Paenibacillus terrae MH72 are AJ318917, AJ318907 and AJ318910, respectively.

A table of the substrate utilization profiles of earthworm-gut isolates and transmission electron micrographs of strains ED5^T and MH72 are available as supplementary material in IJSEM Online.

Present address: Labor L+S AG, Mangelsfeld 4, 97708 Bad Bocklet, Germany.

Present address: Department of Microbiology, University of Aarhus, 8000 Aarhus, Denmark.

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