Tsuneo Shiba

Roseobacter litoralis gen. nov., sp. nov., and Roseobacter denitrificans sp. nov., Aerobic Pink-Pigmented Bacteria which Contain Bacteriochlorophyll a

Summary The new genus Roseobacter , with the two new species Roseobacter litoralis and Roseobacter denitrificans , is proposed for certain aerobic heterotrophs containing bacteriochlorophyll a. R. denitrificans was previously called Erythrobacter sp. OCh114. Members of Roseobacter contain spheroidenone as a major carotenoid, do not synthesize bacteriochlorophyll anaerobically, but show aerobic phototrophic activity. Cells are ovoid or rodshaped, motile by subpolar flagella, and require biotin, nicotinic acid, thiamine and Na + ion. In vivo absorption spectra show a large band at 805–7 nm, and a smaller band at 868–73 nm in the near infrared region. R. denitrificans dissimilatorily reduces nitrate, whereas R. litoralis does not.

Similarity of Tetracycline Resistance Genes Isolated from Fish Farm Bacteria to Those from Clinical Isolates

ABSTRACT Tetracycline-resistant (Tetr) bacteria were isolated from fishes collected at three different fish farms in the southern part of Japan in August and September 2000. Of the 66 Tetr gram-negative strains, 29 were identified as carrying tetB only. Four carried tetY, and another four carried tetD. Three strains carried tetC, two strains carried tetB and tetY, and one strain carried tetC and tetG. Sequence analyses indicated the identity in Tetr genes between the fish farm bacteria and clinical bacteria: 99.3 to 99.9% for tetB, 98.2 to 100% for tetC, 99.7 to 100% for tetD, 92.0 to 96.2% for tetG, and 97.1 to 100% for tetY. Eleven of the Tetr strains transferred Tetr genes by conjugation to Escherichia coli HB-101. All transconjugants were resistant to tetracycline, oxycycline, doxycycline, and minocycline. The donors included strains of Photobacterium, Vibrio, Pseudomonas, Alteromonas, Citrobacter, and Salmonella spp., and they transferred tetB, tetY, or tetD to the recipients. Because NaCl enhanced their growth, these Tetr strains, except for the Pseudomonas, Citrobacter, and Salmonella strains, were recognized as marine bacteria. Our results suggest that tet genes from fish farm bacteria have the same origins as those from clinical strains.

Erythrobacter longus gen. nov., sp. nov., an Aerobic Bacterium Which Contains Bacteriochlorophyll a

Four orange-pigmented and seven pink-pigmented strains of bacteria which contained bacteriochlorophyll a were isolated from high-tidal seaweeds, such as Enteromorpha linza (L.) J. Ag. and Porphyra sp. All of the isolates were gram negative. The orange-pigmented bacteria were rods with parallel sides and rounded ends, and the pink-pigmented bacteria were ovoids and short rods. All were motile by means of subpolar flagella. None of the strains produced growth anaerobically in the light. No growth occurred with an atmosphere containing H2 and CO2. All of these bacteria grew aerobically and utilized glucose, pyruvate, acetate, butyrate, and glutamate as sole organic carbon sources. The best growth occurred on complex media formulated for heterotrophic marine bacteria. Biotin was required. Oxidase and catalase were present. Small amounts of acid were produced from a wide range of carbohydrates under microaerobic conditions. Gelatin was hydrolyzed. The strains which we investigated fell into the following three clusters: cluster A, all of the orange strains; cluster B, three pink strains; and cluster C, four pink strains. The strains of clusters B and C required thiamine and nicotinic acid and were susceptible to streptomycin. Tween 80 was hydrolyzed and phosphatase activity was produced by the strains of clusters A and B. Pantothenate was required only by the strains of cluster C. The guanine-plus-cytosine contents of the deoxyribonucleic acids of these organisms ranged from 60 to 64 mol%. These organisms are recognized here as members of a new genus, Erythrobacter. Although these organisms did not grow phototrophically, the presence of bacteriochlorophyll a indicated that they are most closely related to the Rhodospirillaceae. The type species is Erythrobacter longus, the type strain of which is an orange strain, OCh101 (= IFO 14126).

Heterotrophic bacteria attached to seaweeds

Abstract Viable counts of heterotrophic bacteria attached to the green algae, Monostroma nitidum Wittrock and Enteromorpha linza (Linne) J. Agardh, ranged from 10 4 to 10 6 /cm 2 , and those attached to the red alga Porphyra suborbiculata Kjellman from 10 3 to 10 4 /cm 2 . These bacterial populations were larger than those attached to the brown alga Eisenia bicyclis (Kjellman) setchell ranging from 10 1 to 10 4 /cm 2 . The bacterial populations in the environmental sea water. Nabem Inlet and Otsuchi Bay (Japan), were 10 3 /ml. Orange and yellow pigmented bacteria were predominant on the green and red algae, but not in the bacterial populations of the brown alga and the sea water. Most of the pigmented bacteria were identified as belonging to the Flavobacterium-Cytophaga group. A beneficial relationship was suggested between the green algae and the pigmented bacteria. Proportions of Vibrionaceae were small on the green algae.

Distribution of bacteriochlorophylla in species of the genusAcidiphilium

Bacteriochlorophyll (Bchl)a was found in strains of the strictly aerobic acidophilic heterotrophic bacteria:Acidiphilium rubrum, A. angustum, andA. cryptum. Absorption spectra of the cell-free extracts showed a large peak at 865 nm and a small peak around 802 nm. Anaerobic growth was not observed in either the light or the dark. Bchla was not detected in eitherA. organovorum orA. facilis. Bchla contents were less than ca. 0.7 nmol/mg dry cell weight, being variable among the species.

Isolation of Multidrug-Resistant Stenotrophomonas maltophilia from Cultured Yellowtail (Seriola quinqueradiata) from a Marine Fish Farm

ABSTRACT Six strains of multidrug-resistant Stenotrophomonas maltophilia were isolated from cultured yellowtail. The strains were divided into two clusters based on the 16S rRNA genes, and all of them contained L1 metallo-β-lactamase and L2 β-lactamase genes. Differences in the intercluster divergence between the lactamase genes suggest that horizontal transfer of the genes occurred.

Rubrimonas cliftonensis gen. nov., sp. nov., an aerobic bacteriochlorophyll-containing bacterium isolated from a saline lake.

Phenotypic and phylogenetic studies were performed with two strains (OCh 317T and OCh 318; T = type strain) of aerobic chemoheterotrophic bacteriochlorophyll-containing bacteria isolated from water of a saline lake located on the west coast of Australia. Both strains were Gram-negative, short rods and were motile by means of polar flagella. Catalase, oxidase, nitrate reductase, phosphatase and urease were produced. The cells utilized D-glucose, citrate, glycolate, pyruvate and ethanol. Acids were produced from L-arabinose, D-fructose, D-galactose, D-glucose, D-ribose and D-xylose. The strains could grow in media containing 0.5-7.5% NaCl. Bacteriochlorophyll a was synthesized under aerobic conditions. The results of 16S rRNA gene sequence comparisons revealed that strain OCh 317T represented a new lineage in the alpha-3 group of the class Proteobacteria. Strains OCh 317T and OCh 318 were identified as strains of the same species because of their very similar phenotypic characteristics and their previously described high DNA-DNA homology. Therefore, it was concluded that the two strains should be assigned to a new genus and species, for which the name Rubrimonas cliftonensis is proposed. The type strain is OCh 317T (= JCM 10189T).

Cloning and Sequencing of the Gene Encoding an Aldehyde Dehydrogenase That Is Induced by Growing Alteromonas sp. Strain KE10 in a Low Concentration of Organic Nutrients

ABSTRACT The protein composition of Alteromonas sp. strain KE10 cultured at two different organic-nutrient concentrations was determined by using two-dimensional polyacrylamide gel electrophoresis. The cellular levels of three proteins, OlgA, -B, and -C, were considerably higher in cells grown in a low concentration of organic nutrient medium (LON medium; 0.2 mg of carbon per liter) than cells grown in a high concentration of organic nutrient medium (HON; 200 mg of C liter−1) or cells starved for organic nutrients. In the LON medium, the cellular levels of the Olg proteins were higher at the exponential growth phase than at the stationary growth phase. A sequence of the gene for OlgA revealed that the amino acid sequence had a high degree of similarity to the NAD+-dependent aldehyde dehydrogenases of several bacteria. OlgA, expressed inEscherichia coli, catalyzed the dehydrogenation of acetaldehyde, propionaldehyde, and butyraldehyde. The aldehyde dehydrogenase activity of KE10 was higher in cells growing exponentially in LON medium than in HON. OlgA may be involved in the growth under low-nutrient conditions. The physiological role of OlgA is discussed here.

Similarity in the Structure of tetD-Carrying Mobile Genetic Elements in Bacterial Strains of Different Genera Isolated from Cultured Yellowtail

Structure analysis was performed on the antibiotic-resistance-gene region of conjugative plasmids of four fish farm bacteria.The kanamycin resistance gene, IS26, and tetracycline resistance gene (tetA(D)) were flanked by two IS26s in opposite orientation in Citrobacter sp. TA3 and TA6, and Alteromonas sp. TA55 from fish farm A. IS26-Inner was disrupted with ISRSB101. The chloramphenicol resistance gene, IS26 and tetA (D) were flanked by two IS26s in direct orientation in Salmonella sp. TC67 from farm C. Structures of tetA (D) and IS26 were identical among the four bacteria, but there was no insertion within the IS26-Inner of Salmonella sp. TC67. Horizontal gene transfer between the strains of two different genera in fish farm A was suggested by the structure homologies of mobile genetic elements and antibiotic resistance genes.