A polyphasic taxonomic study was performed on two previously unidentified Arcanobacterium-like Gram-positive strains isolated from harbour seals. Comparative 16S rRNA gene sequencing showed that both bacteria belonged to the genus Arcanobacterium and were most closely related to Arcanobacterium haemolyticum CIP 103370 T (98.4 % 16S rRNA gene sequence similarity), A. canis P6775 T (97.4 %), A. phocae DSM 10002 T (97.4 %), A. pluranimalium M430/94/2 T (95.7 %) and A. hippocoleae CCUG 44697 T (95.5 %). The presence of the major menaquinone MK-9(H 4 ) supported the affiliation of the isolates with the genus Arcanobacterium. The polar lipid profile consisted of major amounts of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, an unidentified phospholipid and two unidentified glycolipids. The major fatty acids were C 16 : 0 , C 18 : 0 , C 18 : 1 v9c and summed feature 5 (comprising C 18 : 2 v6,9c and/or anteiso-C 18 : 0 ). Physiological and biochemical tests clearly distinguished the isolates from other members of the genus Arcanobacterium. Based on the common origin and various physiological properties comparable to those of A. phocae, it is proposed that the isolates are classified as members of a novel species with the name Arcanobacterium phocisimile sp. nov. The type strain is 2698 T (5LMG 27073 T 5CCM 8430 T ).

In a concept study the ability to induce heterogeneous ice formation by Bacterial Ghosts (BGs) from Escherichia coli carrying ice nucleation protein InaZ from Pseudomonas syringae in their outer membrane was investigated by a droplet-freezing assay of ultra-pure water. As determined by the median freezing temperature and cumulative ice nucleation spectra it could be demonstrated that both the living recombinant E. coli and their corresponding BGs functionally display InaZ on their surface. Under the production conditions chosen both samples belong to type II ice-nucleation particles inducing ice formation at a temperature range of between ¡5.6 C and ¡6.7 C, respectively. One advantage for the application of such BGs over their living recombinant mother bacteria is that they are non-living native cell envelopes retaining the biophysical properties of ice nucleation and do no longer represent genetically modified organisms (GMOs).

A yellowish pigmented, Gram-negative, rod-shaped, non-spore-forming bacterium (strain CC-TBT-3 T ), was isolated on marine agar 2216 from a coastal hot spring of Green Island (Lutao), located off Taituang, Taiwan. 16S rRNA gene sequence analysis of strain CC-TBT-3 T showed a relatively low similarity (,95.5 %) to representatives of the genera Novosphingobium, Sphingosinicella and Sphingomonas of the Sphingomonadaceae, with the most related strain being the type strain of Novosphingobium soli. In addition to the relatively low 16S rRNA gene sequence similarity to members of established species, the isolate also showed some unique chemotaxonomic features, including the presence of some glycolipids with unusual chromatographic behaviour. The major components of the polar lipid profile were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid and three unidentified glycolipids. The major respiratory quinone was ubiquinone Q-10. The polyamine pattern was characterized by the triamine sym-homospermidine as a major component. Although the predominant fatty acids were C 18 : 1 v7c and summed feature 3 (C 16 : 1 v7c and/ or iso-C 15 : 0 2-OH), the isolate did not show the typical hydroxyl fatty acids, such as C 14 : 0 2-OH, C 15 : 0 2-OH and C 16 : 0 2-OH, found in members of the genera Novosphingobium, Sphingomonas and Sphingosinicella, but showed instead high amounts of C 18 : 1 2-OH (12.0 %). The DNA G+C content of strain CC-TBT-3 T was 63.4 mol%. 16S rRNA gene sequence, chemotaxonomic and physiological analyses revealed that strain CC-TBT-3 T represents a novel species in a new genus in the family Sphingomonadaceae for which the name Sphingomicrobium lutaoense gen. nov., sp. nov. is proposed; the type strain is of the type species S. lutoaense, CC-TBT-3 T (5DSM 24194 T 5CCM 7794 T ).On the basis of 16S rRNA gene sequence and chemotaxonomic analyses, Takeuchi et al. (2001) proposed a dissection of the former genus Sphingomonas (Yabuuchi et al., 1990(Yabuuchi et al., , 1999, and established the genera Sphingobium, Novosphingobium and Sphingopyxis. Maruyama et al. (2006) added an additional genus, Sphingosinicella, to this group of Abbreviations: pNA, para-nitroanilide; pNP, para-nitrophenyl.

The taxonomy of strain CCUG 55240 T , a Gram-staining-positive, aerobic, endospore-forming bacterium that was isolated from a paper mill, was investigated using a polyphasic approach. In phylogenetic analysis based on 16S rRNA gene sequences, the novel strain was grouped with established members of the genus Paenibacillus and appeared most closely related to the type strains of Paenibacillus chinjuensis (93.7 % sequence similarity), P. elgii (93.7 %) and P. chitinolyticus (93.6 %). The levels of 16S rRNA gene sequence similarity with other species of the genus Paenibacillus, including the type species of the genus, Paenibacillus polymyxa, were all ,93.5 %. The fatty acid profile of strain CCUG 55240 T , which showed a predominance of iso-and anteiso-branched fatty acids, supported the allocation of the strain to the genus Paenibacillus. Unusually high amounts of some iso-branched fatty acids, especially iso-C 15 : 0 and iso-C 16 : 0 , allowed differentiation of strain CCUG 55240 T from the most closely related species of the genus Paenibacillus. The diagnostic diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. The predominant menaquinone was MK-7. The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, an unknown glycolipid, an unknown aminophosphoglycolipid and an unknown phospholipid. Spermidine was the major polyamine. The results of some physiological and biochemical tests also allowed the phenotypic differentiation of strain CCUG 55240 T from the most closely related recognized species. On the basis of the phylogenetic, phenotypic and molecular evidence, strain CCUG 55240 T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus chartarius sp. nov. is proposed. The type strain of the novel species is CCUG 55240 T (5CCM 7759 T ).

The Bordetella species are Gram-negative bacterial pathogens that colonizes mammalian respiratory tract causing respiratory diseases in humans and animals. B. bronchiseptica causes clinical conditions in many mammals including immunocompromised humans. Using the dog model of respiratory infection, it has been shown in this study that a newly developed B. bronchiseptica Bacterial Ghost (BbBG) vaccine exhibited significant protection in the face of a severe pathogenic bacterial challenge in seronegative dogs. The protein E -specific lysis mechanism was used to produce BbBGs. Bacterial Ghosts (BGs) are the empty cell envelope of Gram-negative bacterium. They are genetically processed to form a microscopic hole in their membrane, through which all the cytoplasmic contents are expelled leaving behind intact empty bacterial shells. Due to the intact surface structures of BGs, they offer the safety of inactivated but efficacy of live attenuated vaccines. In this study, seronegative dogs were vaccinated subcutaneously (s/c) with two different doses of a newly developed BbBG vaccine [lower 10 ∧ 5 (BbBG – 5) and higher 10 ∧ 7 (BbBG – 7)] on day 0 and 21. The animals were challenged (by aerosol) with virulent live B. bronchiseptica strains 41 days after first vaccination. The dogs vaccinated s/c with BbBG – 7 vaccine had significantly lower spontaneous coughing scores ( P = 0.0001) than dogs in negative control group. Furthermore, the tested BbBG – 7 vaccine was equivalent to the positive control vaccine Bronchicine CAe in terms of safety and efficacy. For the first time, we report the successful use of liquid formulated BGs vaccines in animal studies. Earlier reported studies using BGs vaccines were performed with resuspended freeze-dried BGs preparations.