Orbus sasakiae sp. nov., a bacterium isolated from the gut of the butterfly Sasakia charonda, and emended description of the genus Orbus

Abstract

Orbus sasakiae sp. nov., a bacterium isolated from the gut of the butterfly Sasakia charonda, and emended description of the genus Orbus Joon Yong Kim, Jina Lee, Na-Ri Shin, Ji-Hyun Yun, Tae Woong Whon, Min-Soo Kim, Mi-Ja Jung, Seong Woon Roh, Dong-Wook Hyun and Jin-Woo Bae ). An emended description of the genus Orbus is provided.Orbus, a genus in the class Gammaproteobacteria, was first introduced by Volkmann et al. (2010). At the time of writing, Orbus hercynius is the only recognized species within this genus and was originally isolated from the faeces of wild boars in Germany. The major fatty acids of the genus Orbus are monounsaturated, even-numbered, straight-chain C 18 (C 18 : 1 v7c) and saturated, even-numbered, straight-chain C 16 (C 16 : 0 ) components (Volkmann et al., 2010). Here, we isolated an Orbus-like strain, designated C7 T , from butterfly gut. Based on taxonomic characterization and phylogenetic analysis, we propose that strain C7T represents a novel species belonging to the genus Orbus. Strain C7T was isolated from the gut of the butterfly Sasakia charonda in South Korea. Cells were isolated by the standard dilution-plating method and were incubated under aerobic conditions at 20 u C on trypticase soy agar (TSA; Difco). To obtain pure cultures, a single colony was repeatedly transferred. Gram reactions were performed using a Gram staining kit (bioMérieux) according to the manufacturer's instructions. Cell morphology and Gram staining of strain C7T were observed using a light microscope (ECLIPSE 50i; Nikon). Motility was examined by the method of Tittsler & Sandholzer (1936). Growth under anaerobic conditions was monitored after 7 days incubation in an anaerobic chamber (N 2 /H 2 /CO 2 , 90 : 5 : 5) at 37 u C on TSA medium. The growth of strain C7 T on CIN agar medium (CIN; BBL) was tested. Based on the experiments described above, strain C7T was determined to be coccoid (0.5-1.0 mm in diameter), Gram-stain-negative, facultatively anaerobic and non-motile. Growth of strain C7T was observed on CIN agar. All experiments were performed in triplicate.Growth at different temperatures (4, 15, 20, 25, 30, 37, 45 and 65 u C), pH values (pH 4.0-11.0, at intervals of 1.0 pH unit) and NaCl concentrations (0, 1, 2, 3, 4, 5, 8, 10, 12 and 15 %, w/v) was tested in trypticase soy broth (TSB; Difco). pH was adjusted by adding the following buffers: 10 mM MES (C 6 H 13 NO 4 S) (for pH 4, 5 and 6), 10 mM TAPS (C 7 H 14 NO 6 S) (for pH 7, 8 and 9) or 10 mM Na 2 HPO 4 (for pH 10 and 11). The strains were cultivated in TSB at 20 u C for all experiments, unless stated otherwise. After incubation for 24 h, 48 h or 7 days, the turbidity of the cultures was measured as the optical density at 600 nm (OD 600 ) using a spectrophotometer (SYNERGY MX; BioTek). Strain C7T grew with 0-5 % (w/v) NaCl, atAbbreviations: PE, phosphatidylethanolamine; PG, phosphatidylglycerol.

Frischella perrara gen. nov., sp. nov., a gammaproteobacterium isolated from the gut of the honeybee, Apis mellifera

“…Strain PEB0191 T was grown in peptoneyeast-glucose broth at 35 u C for 20-24 h under anaerobic conditions, corresponding to the late-exponential phase of physiological growth. Similar to other members of the family Orbaceae (Kwong & Moran, 2013;Kim et al, 2013;Volkmann et al, 2010), palmitic acid (C 16 : 0 ) and cisvaccenic acid (C 18 : 1 v7c and/or C 18 : 1 v6c) were found to be the major fatty acids of strain PEB0191 T (Table 1). Respiratory quinones were extracted and identified as described by Kwong & Moran (2013); the major quinone detected in PEB0191 T was ubiquinone 8, which is also consistent with other members of the family Orbaceae.…”

“…Bars, 5 mm. were conducted with the API50 CH method (bioMérieux) (Kim et al, 2013); fermentation data for PEB0191 T and G. apicola wkB1 T were obtained in this study under the same laboratory conditions using the method described by MacFaddin (2000). 2, Not detected.…”

“…2010). Recently, a second species of the genus Orbus has been described; Orbus sasakiae C7 T was isolated from the gut of the butterfly Sasakia charonda (Kim et al, 2013). Sequencing-based methods revealed the existence of additional taxa within the family Orbaceae.…”

Draft Genome Sequence of Commensalibacter papalotli MX01, a Symbiont Identified from the Guts of Overwintering Monarch Butterflies

“…Overwintering monarchs reduce their metabolism and limit their feeding. Few studies have described gut bacteria in butterflies (1–8), and there are no studies on the molecular identification of symbionts from monarch butterflies.…”

Paenalcaligenes hermetiae sp. nov., isolated from the larval gut of Hermetia illucens (Diptera: Stratiomyidae), and emended description of the genus Paenalcaligenes

“…BSF larvae are known to dramatically reduce food waste and to convert the nutrients from food waste and manure into insect larval biomass (Kim et al, 2008;Diener et al, 2009). Thus, the BSF larva itself is a potential high protein and high energy feed as well as a resource for isolation of novel species of bacteria, archaea, yeasts and protists from insect guts of adults and larvae (Breznak, 1982;Brauman et al, 2001;Hongoh et al, 2003;Schmitt-Wagner et al, 2003;Ohkuma, 2008;Kudo, 2009;Jeon et al, 2011;Kim et al, 2013). …”