Preparation of a Stable Mycobacterial Tween Hydrolysis Test Substrate

Kilburn, J. O.; O'Donnell, K. F.; Silcox, V. A.; David, H. L.

doi:10.1128/aem.26.5.826-826.1973

Cited by 12 publications

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“…Growth ability on MacConkey agar plates was tested using standard MacConkey agar (Scharlau Microbiology) at 18 °C for seven days. Hydrolysis of casein, gelatin and starch was determined as described by Smibert and Krieg (1994) and hydrolysis of Tween 80 was determined as described by Kilburn et al (1973). Citrate utilization was tested at 18 °C for up to 7 days (Smibert & Krieg, 1994).…”

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Paenibacillus darwinianus sp. nov., isolated from gamma-irradiated Antarctic soil

Dsouza

Taylor

Ryan

et al. 2014

International Journal of Systematic and Evolutionary Microbiology

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A novel bacterium, strain Br T , was isolated from gamma-irradiated soils of the Britannia drift, Lake Wellman Region, Antarctica. This isolate was rod-shaped, endospore forming, Gram-stainvariable, catalase-positive, oxidase-negative and strictly aerobic. Cells possessed a monotrichous flagellum. Optimal growth was observed at 18 6C, pH 7.0 in PYGV or R2A broth. The major cellular fatty acid was anteiso-C 15 : 0 (63.4 %). Primary identified lipids included phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. Total phospholipid was 60 % (w/w) of the total lipid extract. MK-7 was the dominant isoprenoid quinone. The genomic DNA G+C content was 55.6 mol%. Based on 16S rRNA gene sequence similarity, strain Br T clusters within the genus Paenibacillus with similarity values ranging from 93.9 to 95.1 %. Phylogenetic analyses by maximum-likelihood, maximum-parsimony and neighbourjoining methods revealed that strain Br T clusters with Paenibacillus daejeonensis (AF290916), Paenibacillus tarimensis (EF125184) and Paenibacillus pinihumi (GQ423057), albeit with weak bootstrap support. On the basis of phenotypic, chemotaxonomic and phylogenetic characteristics, we propose that strain Br T represents a novel species, Paenibacillus darwinianus sp. nov. The type strain is Br T (5DSM 27245 T 5ICMP 19912 T ).The genus Paenibacillus belonging to family Paenibacillaceae was first resolved from members of rRNA group 3 bacilli by Ash et al. (1993) based on comparative 16S rRNA gene sequence analysis. Members of the genus Paenibacillus are characterized as rod-shaped, Gram-positive or variable (depending on growth phase), endospore-forming, aerobic or facultatively anaerobic bacteria, possessing anteiso-C 15 : 0 as the major cellular fatty acid and unsaturated menaquinone with seven units (MK-7) as the predominant isoprenoid quinone, and having a genomic DNA G+C content ranging between 40 and 54 mol% (Ash et al., 1993;Shida et al., 1997). The genus Paenibacillus contains several species that have been isolated from a wide range of cold environments including sediment (Montes et al., 2004) and soil (Marshall & Ohye, 1966; Rodríguez-Díaz et al., 2005) from the Antarctic Peninsula, a cold spring in China (Tang et al., 2011), soil on the Kafni glacier, Himalayas (Kishore et al., 2010), Alaskan soil (Nelson et al., 2009) and a peat bog in Northern China (Ming et al., 2012). In this study we investigated the taxonomic status of strain Br T isolated from Antarctic soils which were exposed to gamma irradiation, using a combination of physiological, biochemical, chemotaxonomic and phylogenetic techniques. Based on these analyses we propose strain Br T as a representative of a novel species, Paenibacillus darwinianus sp. nov.Soil samples on the Britannia drift, Lake Wellman Region (79 u 559 16.20 S 156 u 559 30.70 E) in the south-eastern Darwin Mountains, Antarctica, were collected for the isolation of bacteria (Aislabie et al., 2012 Colony morphology was observed on PYGV gellan gum plates after culturing for 4 days at...

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confidence: 99%

Paenibacillus darwinianus sp. nov., isolated from gamma-irradiated Antarctic soil

Dsouza

Taylor

Ryan

et al. 2014

International Journal of Systematic and Evolutionary Microbiology

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“…Assays of amylase activity (starch) on nutrient agar supplemented with potato starch, according to the FDA Bacteriological Analytical Manual, yielded negative results. Furthermore, Tween 80 hydrolysis (lipase) by N384 T was shown to be negative by two separate methods (Kilburn et al , 1973; Oliver et al , 1986). Casein hydrolysis by N384 T was negative on heart infusion agar containing 1 % NaCl and 1 % skimmed milk, as described by Hülsmann et al (2003).…”

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Vibrio caribbeanicus sp. nov., isolated from the marine sponge Scleritoderma cyanea

Hoffmann

Monday

Allard

et al. 2012

International Journal of Systematic and Evolutionary Microbiology

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A Gram-negative, oxidase-positive, catalase-negative, facultatively anaerobic, motile, curved rodshaped bacterium, strain N384 T , was isolated from a marine sponge (Scleritoderma cyanea; phylum Porifera) collected from a depth of 795 feet (242 m) off the west coast of Curaç ao. On the basis of 16S rRNA gene sequencing, strain N384 T was shown to belong to the genus Vibrio, most closely related to Vibrio brasiliensis LMG 20546 T (98.8 % similarity), Vibrio nigripulchritudo ATCC 27043 T (98.5 %), Vibrio tubiashii ATCC 19109 T (98.6 %) and V. sinaloensis DSM 21326 T (98.2 %). The DNA G+C content of strain N384 T was 41.6 mol%. An analysis of concatenated sequences of five genes (gyrB, rpoA, pyrH, mreB and ftsZ; 4068 bp) demonstrated a clear separation between strain N384 T and its closest neighbours and clustered strain N384 T into the 'Orientalis' clade of vibrios. Phenotypically, the novel species belonged to the arginine dihydrolase-positive, lysine decarboxylase-and ornithine decarboxylase-negative (A+/L"/O") cluster. The novel species was also differentiated on the basis of fatty acid composition, specifically that the proportions of iso-C 13 : 0 , iso-C 15 : 0 , C 15 : 0 , iso-C 16 : 0 , C 16 : 0 , iso-C 17 : 0 , C 17 : 1 v8c and C 17 : 0 were significantly different from those found in V. brasiliensis and V. sinaloensis. The results of DNA-DNA hybridization, average nucleotide identity and physiological and biochemical tests further allowed differentiation of this strain from other described species of the genus Vibrio. Collectively, these findings confirm that strain N384 T represents a novel Vibrio species, for which the name Vibrio caribbeanicus sp. nov. is proposed, with the type strain N384 T (5ATCC BAA-2122 T 5DSM 23640 T ).The family Vibrionaceae is a member of the class Gammaproteobacteria and consists of the genera Vibrio, Photobacterium, Salinivibrio, Grimontia, Enterovibrio and Aliivibrio (Chimetto et al., 2011). At the time of writing, the genus Vibrio consisted of 89 species (http://www.vibriobiology.net). Some vibrios, such as Vibrio cholerae, V. parahaemolyticus and V. vulnificus, are well-known human pathogens (FDA, 2004). Generally, Vibrio species are halophilic, mesophilic and chemo-organotrophic in nature and have a facultatively fermentative metabolism (Thompson et al., 2004). Typically, vibrios are inhabitants of aquatic environments that show a remarkable degree of biodiversity, persisting in a variety of geographical locales and eukaryotic hosts, including corals, molluscs, sponges and zooplankton (Thompson et al., 2004).Sponges can maintain diverse symbiotic microbe populations that differ in composition from the microbial communities Abbreviations: ANI, average nucleotide identity; HGT, horizontal gene transfer; ILD, incongruence length difference; ML, maximum-likelihood; MLSA, multilocus sequence analysis; MP, maximum parsimony. The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene, topA, ftsZ, gapA, gyrB, mreB, pyrH, recA and rpoA gene sequences derived in this study are H...

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“…E. coli DH5 α cells were used as a negative control. The streptococcus thermophiles activity of the bacteria were assessed using the Tween 80 hydrolysis test that was performed as previously reported [ 24 ].…”

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confidence: 99%

Identification of a NewAlcaligenes faecalisStrain MOR02 and Assessment of Its Toxicity and Pathogenicity to Insects

Quiroz-Castañeda

Mendoza-Mejía

Obregón-Barboza

et al. 2015

BioMed Research International

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We report the isolation of a bacterium from Galleria mellonella larva and its identification using genome sequencing and phylogenomic analysis. This bacterium was named Alcaligenes faecalis strain MOR02. Microscopic analyses revealed that the bacteria are located in the esophagus and intestine of the nematodes Steinernema feltiae, S. carpocapsae, and H. bacteriophora. Using G. mellonella larvae as a model, when the larvae were injected with 24,000 CFU in their hemocoel, more than 96% mortality was achieved after 24 h. Additionally, toxicity assays determined that 1 μg of supernatant extract from A. faecalis MOR02 killed more than 70% G. mellonella larvae 96 h after injection. A correlation of experimental data with sequence genome analyses was also performed. We discovered genes that encode proteins and enzymes that are related to pathogenicity, toxicity, and host/environment interactions that may be responsible for the observed phenotypic characteristics. Our data demonstrates that the bacteria are able to use different strategies to colonize nematodes and kill insects to their own benefit. However, there remains an extensive group of unidentified microorganisms that could be participating in the infection process. Additionally, a nematode-bacterium association could be established probably as a strategy of dispersion and colonization.

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Preparation of a Stable Mycobacterial Tween Hydrolysis Test Substrate

Abstract: A Tween 80 hydrolysis test substrate was found to be stable at room temperature for at least 6 months.

Cited by 12 publications

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Paenibacillus darwinianus sp. nov., isolated from gamma-irradiated Antarctic soil

Paenibacillus darwinianus sp. nov., isolated from gamma-irradiated Antarctic soil

Vibrio caribbeanicus sp. nov., isolated from the marine sponge Scleritoderma cyanea

Identification of a NewAlcaligenes faecalisStrain MOR02 and Assessment of Its Toxicity and Pathogenicity to Insects

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