The major fatty acids were C15:0 iso 2-OH and/or C16:1ω7, C16:0, C18:1ω7 and C14:0. Based on the polyphasic evidence
presented here, it can be concluded that strains DY05T and 47666-1 belong to the same novel species of the genus Vibrio, for which the name Vibrio owensii Regorafenib in vitro sp. nov. is proposed. The type strain is DY05T (=JCM 16517T=ACM 5300T). Recently, the number of bacterial species of the genus Vibrio (Farmer et al., 2005) has increased noticeably. Currently, the Harveyi clade (Sawabe et al., 2007) includes eight species: Vibrio harveyi, Vibrio campbellii, Vibrio rotiferianus, Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio mytili, Vibrio natriegens and the newly described Vibrio azureus (Yoshizawa et al., 2009). Among this group, V. harveyi has been recognized as the most significant pathogen of marine-reared fish and crustaceans (Karunasagar et al., 1994; Zhang & Austin, 2000), and several studies have reported infections by this species in molluscs and corals (Nishimori et al., 1998; Sutherland selleck chemicals et al., 2004). More recently, however, molecular analyses revealed that some disease causing strains of V. campbellii have been misidentified as V. harveyi, underestimating the significance of the former species as an aquaculture
pathogen (Gómez-Gil et al., 2004). Here, we describe the physiological, chemotaxonomic and phylogenetic characteristics of two bacterial strains pathogenic to cultured crustaceans sharing the highest Dimethyl sulfoxide 16S rRNA gene sequence identities with V. harveyi, V. campbellii and V. rotiferianus. The strain 47666-1 was isolated from diseased Penaeus monodon larvae in a commercial prawn hatchery in North Queensland, Australia, and subsequently shown to be highly virulent to prawn larvae (Harris, 1993; Pizzuto & Hirst, 1995). Similarly, strain DY05T was isolated from diseased larvae of the ornate spiny lobster Panulirus ornatus in the Tropical Aquaculture Facility of the Australian Institute of Marine Science (AIMS), North Queensland, Australia, and subsequently shown to be highly virulent to lobster larvae (unpublished data). Bacteria (strains DY05T and 47666-1, V. harveyi
LMG 4044T, V. campbellii LMG 11216T, V. rotiferianus LMG 21460T and V. rotiferianus CAIM 994) were cultured on thiosulphate–citrate–bile–salts–sucrose (TCBS) agar and marine agar (MA) at 28 °C. Stock cultures were maintained frozen at −80 °C in either marine broth (MB) with 30% v/v glycerol or in Microbank™ cryovials (Pro-Lab Diagnostics). For morphology and physiology studies, cells were grown for 24–48 h at 28 °C on MA or in MB. Gram staining was performed using a Gram stain kit (Becton Dickinson, BD) according to the manufacturer’s instructions. Cell morphology, size and motility were determined by light microscopy (CX31, Olympus). Luminescence was observed in the dark and measured using a 1420 Wallac Multilabel Counter (Perkin Elmer) at 4-h intervals.