Corynebacterium doosanense sp. nov., isolated from activated sludge

Abstract

The taxonomic position of a Gram-positive, non-motile, non-spore-forming coryneform, isolated from activated sludge and designated strain CAU 212 T , was investigated using a polyphasic approach. Cellular morphology, biochemical tests and chemotaxonomic investigations revealed that strain CAU 212 T had the characteristics of the genus Corynebacterium. Comparative 16SrRNA gene sequence analysis showed that the organism formed a hitherto-unknown subline within the genus Corynebacterium. Sequence divergence values of more than 4.3 % from recognized Corynebacterium species, together with phenotypic differences, showed that the bacterium represents a previously unrecognized member of the genus Corynebacterium, for which the name Corynebacterium doosanense sp. nov. is proposed. The type strain is CAU 212The genus Corynebacterium was established by Lehmann & Neumann (1896) and represents one of the largest groups within the Actinobacteria. At the time of writing, Corynebacterium comprised 95 recognized taxa. In recent years, many of the recognized species within this genus originated from clinical specimens of humans (Hall et al., 2003;Renaud et al., 2007;Yassin, 2007;Yassin & Siering, 2008), animals (Goyache et al., 2003a, b; Fernández-Garayzábal et al., 2003Collins et al., 2004;Vela et al., 2006) and environments (Yassin et al., 2003; Chen et al., 2004;Feurer et al., 2004). Until now, however, there have been relatively few reports of corynebacteria from activated sludge. During the course of routine screening of bacteria for industrial purposes, a Corynebacterium-like strain, designated CAU 212 T , was isolated from activated sludge from the wastewater treatment plant in Yeongdeuk-gun, Republic of Korea.The isolation procedure for strain CAU 212 T followed that of Gordon & Mihm (1962) by using glucose-yeast extract agar (GYEA; comprising per litre: 10 g yeast extract, 10 g glucose and 15 g agar) supplemented with 50 mg cycloheximide l 21 and 20 mg nalidixic acid l 21. An activated sludge sample was diluted with sterilized distilled water and appropriate dilutions were spread onto the GYEA medium and incubated aerobically for 3 days at 30 u C. An opaque, yellow, low-convex bacterial colony with a diameter of 1-2 mm was subcultured on sheep blood agar plates (Asan Pharm Co.). The pure culture of CAU 212 T was preserved in 25 % (v/v) glycerol at 270 u C. The cell morphology of colonies was observed at different incubation times, every 24 h for 3 days, with a microscope following Gram staining. Strain CAU 212T was characterized biochemically using the API Coryne, API 20 Strep and API ZYM kits (bioMérieux) according to the manufacturer's instructions. The ChristieAtkins-Munch-Petersen (CAMP) test with Staphylococcus aureus was performed as described by von Graevenitz & Funke (1996). Cell-wall murein was prepared by mechanical disruption of cells and complete acid hydrolysates were analysed as described by Schleifer & Kandler (1972). Cellular fatty acid methyl esters were extracted by acid methanolysis (Minnikin et al.,...

Genome-Based Taxonomic Classification of the Phylum Actinobacteria

“…The description is as before (Lee et al, 2009 ) with the following modification. The G+C content of the type-strain genome is 66.9%, its approximate size 2.65 Mbp, its GenBank deposit SAMN02256506 .…”

The Family Corynebacteriaceae

“…A selective medium for the isolation of Corynebacterium urealyticum from urine and skin samples of hospitalized patients has been described (Zapardiel et al 1998). Specific media have been designed and applied for the isolation of environmental corynebacteria from activated sludge (Corynebacterium doosanense) (Lee et al 2009), the anode of a wastewater-fed microbial fuel cell (Corynebacterium humireducens) (Wu et al 2011), saline soil (Corynebacterium halotolerans) (Chen et al 2004), and coastal sediment (Corynebacterium marinum) (Du et al 2010).…”

“…Other Corynebacterium species have been described as being recovered from food products or from the environment, such as soil, water, marine ecosystems, or plant surfaces: Corynebacterium callunae from heather (Yamada and Komagata 1972;Bernard and Funke 2012); Corynebacterium casei from the surface of smear-ripened cheeses Mounier et al 2005); Corynebacterium deserti from a mixed sand sample collected in a desert in the west of China (Zhou et al 2012); Corynebacterium doosanense from activated sludge taken from a wastewater treatment plant in Yeongdeuk-gun, Republic of Korea (Lee et al 2009); Corynebacterium efficiens from soil, vegetables, and onion bulbs (Fudou et al 2002); Corynebacterium flavescens from dairy products (Bernard and Funke 2012); Corynebacterium glaucum from a cosmetic dye (Yassin et al 2003); Corynebacterium glutamicum from soil contaminated with bird feces, sewage, manure, vegetables, and fruits (Abe et al 1967;Liebl 2005); Corynebacterium halotolerans from a saline soil sample that was collected in Xinjiang Province, China (Chen et al 2004); Corynebacterium humireducens from the anode of a microbial fuel cell fed with artificial wastewater (Wu et al 2011); Corynebacterium lubricantis from a coolant lubricant ); Corynebacterium marinum from coastal sediment close to a coal-fired power station in Qingdao, China (Du et al 2010); Corynebacterium maris from the mucus of the coral Fungia granulosa from northern Red Sea, Gulf of Eilat, Israel (Ben-Dov et al 2009); Corynebacterium nuruki from nuruk which is a Korean alcohol fermentation starter (Shin et al 2011a); Corynebacterium terpenotabidum from soil (Takeuchi et al 1999); and Corynebacterium variabile from the surface of smear-ripened cheeses Mounier et al 2005).…”

Corynebacterium guangdongense sp. nov., isolated from a contaminated plate

“…At the time of writing, the genus comprises more than 100 members characterized by Grampositive, asporogenous, rod-shaped and containing menaquinone MK-8(H 2 ) and/or MK-9(H 2 ) as isoprenoid quinones. They were isolated from various sources, such as birds (Fern andez-Garayz abal et al, 2004;K€ ampfer et al, 2015), milk (Wiertz et al, 2013), pen shell (Kim et al, 2015), activated sludge (Lee et al, 2009), human clinical samples (Otsuka et al, 2005;Riegel et al, 2006), etc. Here we described a novel member of the genus Corynebacterium, designated strain S01 T , which was recovered from a plate contaminated from an unknown source in our laboratory, based on phenotypic, phylogenetic and chemotaxonomic approaches.…”