Rahnella perminowiae sp. nov., Rahnella bonaserana sp. nov., Rahnella rivi sp. nov. and Rahnella ecdela sp. nov., isolated from diverse environmental sources, and emended description of the genus Rahnella.

Bacteria isolated from onion bulbs suffering from bacterial decay in the United States and Norway were previously shown to belong to the genus Rahnella based on partial housekeeping gene sequences and/or fatty acid analysis. However, many strains could not be assigned to any existing Rahnella species. Additionally, strains isolated from creek water and oak as well as a strain with bioremediation properties were assigned to Rahnella based on partial housekeeping gene sequences. The taxonomic status of these 21 strains was investigated using multilocus sequence analysis, whole genome analyses, phenotypic assays and fatty acid analysis. Phylogenetic and phylogenomic analyses separated the strains into five clusters, one of which corresponded to Rahnella aceris. The remaining four clusters could be differentiated both genotypically and phenotypically from each other and existing Rahnella species. Based on these results, we propose the description of four novel species: Rahnella perminowiae sp. nov. (type strain SL6T=LMG 32257T=DSM 112609T), Rahnella bonaserana sp. nov. (H11bT=LMG 32256T=DSM 112610T), Rahnella rivi sp. nov. (FC061912-KT=LMG 32259T=DSM 112611T) and Rahnella ecdela sp. nov. (FRB 231T=LMG 32255T=DSM 112612T).

Rahnella laticis sp. nov. and Rahnella contaminans sp. nov., and emended description of the genus Rahnella.

Taxonomic positions of six isolates, which were recovered from two different environments in Jeju, Republic of Korea, were examined by a polyphasic analysis. Cells of the isolates were Gram-reaction-negative, facultatively anaerobic, motile and rod-shaped and showed growth at 4-30 °C, pH 4.0-9.0 and with 0-6 (w/v) NaCl. In phylogenomic analysis based on 92 single-copy core genes, it was shown that the isolates belonged to the genus Rahnella and formed three distinct sublines within the genus. The isolates shared 16S rRNA gene sequence similarities of 97.9-100 % with one another. The isolates contained ubiquinone-8 was as the major isoprenoid quinone. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and an unidentified aminophospholipid. The predominant fatty acids were C16 : 0 and C17 : 0 cyclo. The G+C content of their genomic DNA was 52.8-53.1 %. Average nucleotide identity and digital DNA-DNA hybridization values supported that strains SAP-17T and Lac-M11T represented two new species of the genus Rahnella, whereas strain SAP-10 was a strain of Rahnella victoriana. Based on the results obtained here, Rahnella laticis sp. nov. (type strain SAP-17T=KCTC 72960T=NBRC 114723T=CCM 9079T) and Rahnella contaminans sp. nov. (type strain Lac-M11T=KACC 21743T=NBRC 114406T) are proposed. Also, an emended description of the genus Rahnella is given on the basis of our physiological and chemotaxonomic results.

Septic Shock Caused by Rahnella aquatilis Bacteremia in an Immunocompetent Adult.

BACKGROUND Rahnella aquatilis is a facultatively anaerobic, gram-negative rod bacterium commonly found in freshwater. There are few cases of bacteremia caused by Rahnella aquatilis in the literature and even fewer cases reported of it causing sepsis in immunocompetent individuals. In this case report, we present a rare case of an immunocompetent individual who developed sepsis secondary to bacteremia caused by Rahnella aquatilis. CASE REPORT A 37-year-old immunocompetent man with cerebral palsy and chronic enterocutaneous fistulas, with an indwelling peripherally inserted central catheter (PICC) line for total parenteral nutrition (TPN), presented to the emergency department with complaints of increased enteric drainage from his fistula, rigors, and subjective fevers following a mechanical fall, which occurred approximately 1 week before. The day following admission, the patient developed septic shock and was transferred to the intensive care unit for vasopressor support. He was given intravenous cefepime and metronidazole for empiric therapy. Blood cultures grew Rahnella aquatilis, and antibiotic therapy was de-escalated to monotherapy with intravenous ceftriaxone. The patient's condition stabilized, his PICC line was replaced, and he was successfully discharged, and continued on outpatient antibiotic therapy with ceftriaxone. CONCLUSIONS This case report represents a novel presentation of septic shock secondary to bacteremia caused by a gram-negative rod bacterium, Rahnella aquatilis, in an immunocompetent host dependent on TPN via a PICC line. This case also demonstrates that Rahnella aquatilis can be susceptible to and treated successfully with intravenous ceftriaxone. Bacteremia caused by Rahnella aquatilis can cause a swift, aggressive decompensation and should be treated with antibiotics immediately.

Rahnella aceris sp. nov., isolated from sap drawn from Acer pictum.

A Gram-reaction-negative, facultatively anaerobic bacterium, designated SAP-19T, was isolated from sap extracted from Acer pictum in Mt. Halla in Jeju, Republic of Korea and its taxonomic statue was investigated by a polyphasic approach including genome- and 16S rRNA gene-based phylogenetic analyses. Cells were motile, short rods and showed growth at 20-30 °C, pH 4-9 and 0-6% (w/v) NaCl. The whole genome- and 16S rRNA gene-based phylogenetic analyses exhibited that strain SAP-19T belongs to the genus Rahnella and forms a tight cluster with Rahnella aquatilis. The isolate shared average nucleotide identity of 92.7% and 16S rRNA gene sequence similarity of 99.6% with the type strain of Rahnella aquatilis. The polar lipids contained phosphatidylethanolamine, an unidentified aminophospholipid and an unidentified lipid. The major isoprenoid quinone was Q-8. The predominant fatty acids were C16:0 and C17:0cyclo. The G + C content of the genome was 52.3%. The low average nucleotide identity (92.7%) and digital DNA relatedness (48.6%) values between the isolate and the most closely related strain showed that the isolate can be considered a different genospecies. On the basis of combined data obtained in this study, strain SAP-19T (= KACC 21744T = NBRC 114407T) represents a novel species of the genus Rahnella, for which the name Rahnella aceris sp. nov. is proposed.

The complete genome sequence of Rahnella aquatilis ZF7 reveals potential beneficial properties and stress tolerance capabilities.

Rahnella aquatilis ZF7 is a plant beneficial strain isolated from Sakura tree soil with potential for biocontrol. Here, we present the complete genome sequence of R. aquatilis ZF7, which consists of one 4.49 Mb circular chromosome and a 54-kb plasmid named pRAZF7. Phylogenetic analyses revealed that R. aquatilis ZF7 is much similar to the strains Rahnella sp. Y9602 and R. aquatilis HX2 than others evaluated. In this study, multiple genes encoding functions that likely contribute to plant growth promotion, biocontrol and stress tolerance were identified by comparative genome analyses, including IAA production, phosphate solubilization, antibiotic resistance and formation of Se nanoparticles (SeNPs). In addition, these functions were also confirmed by in vitro experiments. Considering its ability to form SeNPs, strain R. aquatilis ZF7 will contribute to nano-agriculture. Overall, the features of R. aquatilis ZF7 make it a high potential and competitive strain in biocontrol, and the genome data will help further studies on the mechanisms of plant growth promotion and biocontrol.

Rahnella spp. are commonly isolated from onion (Allium cepa) bulbs and are weakly pathogenic.

AIMS: Bacterial decays of onion bulbs have serious economic consequences for growers, but the aetiologies of these diseases are often unclear. We aimed to determine the role of Rahnella, which we commonly isolated from bulbs in the United States and Norway, in onion disease. METHODS AND RESULTS: Isolated bacteria were identified by sequencing of housekeeping genes and/or fatty acid methyl ester analysis. A subset of Rahnella spp. strains was also assessed by multilocus sequence analysis (MLSA); most onion strains belonged to two clades that appear closely related to R. aquatilis. All tested strains from both countries caused mild symptoms in onion bulbs but not leaves. Polymerase chain reaction primers were designed and tested against strains from known species of Rahnella. Amplicons were produced from strains of R. aquatilis, R. victoriana, R. variigena, R. inusitata and R. bruchi, and from one of the two strains of R. woolbedingensis. CONCLUSIONS: Based on binational testing, strains of Rahnella are commonly associated with onions, and they are capable of causing mild symptoms in bulbs. SIGNIFICANCE AND IMPACT OF THE STUDY: While Rahnella strains are commonly found within field-grown onions and they are able to cause mild symptoms, the economic impact of Rahnella-associated symptoms remains unclear.

Environmental Isolate of Rahnella aquatilis Harbors Class 1 Integron.

The paper presents first description of class 1 integron in an environmental strain of Rahnella aquatilis, a rarely isolated Gram-negative bacterium of the family Enterobacteriaceae. The strain was isolated from the Warta river water, Poland. Class 1 integrase gene was detected by a PCR assay. Sequencing of the integron's variable region showed the presence of a dfrA1-aadA1 gene cassette array. The integron was located in a 54-kbp plasmid that was transferable to Escherichia coli J-53 recipient strain in a conjugation assay. The integron-bearing R. aquatilis strain was resistant to aminoglycosides, penicillins, trimethoprim, sulfamethoxazole, and trimethoprim/sulfamethoxazole. This paper confirms that water environment play a major role in the spread of integrons and, consequently, antimicrobial resistance, among bacteria of various genera.

Co-transmission of Rahnella aquatilis between hospitalized patients.

Rahnella aquatilis is an environmental Gram-negative bacillus that is rarely reported as human pathogen, being mainly associated with infections in immunocompromised patients. Herein we describe two cases of R. aquatilis isolates recovered from endotracheal aspirate cultures of different patients in a tertiary hospital located in the city of São Paulo, Brazil. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rDNA gene sequencing were performed to confirm bacterial identification after the isolates being erroneously identified as Pantoea spp. by automated system. Both isolates showed the same PFGE pattern and presented the ß-lactamase encoding gene blaRAHN-1, responsible for resistance to cephalothin. The isolates were susceptible to broad-spectrum cephalosporins, carbapenems, fluoroquinolones, aminoglycosides, and polymyxin B. This report shows the presence and transmission of uncommon bacteria in the nosocomial environment and alerts us about the need for new tools of correct microbiologic diagnosis.

Identification of tetrodotoxin-producing bacteria from goby Yongeichthys criniger.

Toxic goby fish (Yongeichthys criniger) containing tetrodotoxin (TTX), from Zhanjiang, Guangdong province, China, were screened for TTX-producing bacteria. Two toxic bacterial strains were isolated from the liver of Y. criniger and respectively denoted XC3-3 and XL-1. TTX production by the strains was confirmed by mouse bioassay, enzyme-linked immunosorbent assay and high performance liquid chromatography coupled with mass spectrometry. Based on morphological, physiological and biochemical characteristics and 16S rDNA phylogenetic analysis, strain XC3-3 was identified as Enterobacter cloaca and XL-1 was closely related to Rahnella aquatilis. These findings show for the first time that TTX-producing bacteria are symbiotic bacteria in goby and suggest that bacterial strains are at least partially responsible for TTX accumulation in Y. criniger.

Rahnella sp. strain EK12: Cell surface properties and diesel oil biodegradation after long-term contact with natural surfactants and diesel oil.

The changes in cell surface properties of Rahnella sp. strain EK12 and modifications in genetic material after long-term contact with saponins and rhamnolipids, were investigated. Rhamnolipids caused a decrease of hydrophobicity in liquid cultures compared with saponins. On the other hand, in cultures with rhamnolipids, the addition of diesel oil results in a rapid rise of cell surface hydrophobicity. The similar effect was not so significant in the presence of saponins. For the bacteria grown in the presence of saponins or rhamnolipids, but without diesel oil, the ratio of unsaturated to saturated fatty acids decreased, in comparison to the control culture. The differences observed in hydrophobicity, zeta potential and fatty acids profiles, indicated various mechanisms of an interaction between a surfactant and a bacterial cells. The results have also shown an impact of the long-term contact on changes in genetic material of Rahnella sp. strain EK12 cells. Moreover, the presence of saponins led to significant increase of diesel oil biodegradation.

Identification of effective Pb resistant bacteria isolated from Lens culinaris growing in lead contaminated soils.

Soil bacteria are a new phytoremediation system for the removal of heavy metals from soils. In this study, fifteen soil bacteria were isolated from root nodules of lentil growing in heavy metals contaminated soils, particularly by lead. Molecular characterization of the collection showed a large diversity, including Agrobacterium tumefaciens, Rahnella aquatilis, Pseudomonas, and Rhizobium sp. These soil bacteria had a wide range of tolerance to heavy metals. Among them, strains of A. tumefaciens and R. aquatilis tolerated up to 3.35 mM Pb; whereas Pseudomonas tolerated up to 3.24 mM Pb. The inoculation of lentil grown hydroponically with inoculums formed by these efficient and Pb resistant bacteria enhanced plant biomass. The treatment of this symbiosis by 1 mM Pb for 10 days or by 2 mM Pb for 3 days demonstrated that lentil had Pb accumulation capacity and can be considered a Pb accumulator plant, elsewhere, roots accumulated more Pb than shoots, and the inoculation decreased the Pb up take by the plants, suggesting that this symbiosis should be investigated for use in phytostabilization of Pb-contaminated soils. At the same time, a modulation in the antioxidant enzyme activity and a specific duration was required for the induction of the superoxide dismutase (SOD), peroxidase (POX), and ascorbate peroxidase (APX) response and to adapt to Pb stress. These results suggested that these enzymes may be involved in the main mechanism of antioxidative defense in lentil exposed to Pb oxidative stress.

Rahnella victoriana sp. nov., Rahnella bruchi sp. nov., Rahnella woolbedingensis sp. nov., classification of Rahnella genomospecies 2 and 3 as Rahnella variigena sp. nov. and Rahnella inusitata sp. nov., respectively and emended description of the genus Rahnella.

Isolations from oak symptomatic of Acute Oak Decline, alder and walnut log tissue, and buprestid beetles in 2009-2012 yielded 32 Gram-negative bacterial strains showing highest gyrB sequence similarity to Rahnella aquatilis and Ewingella americana. Multilocus sequence analysis (using partial gyrB, rpoB, infB and atpD gene sequences) delineated the strains into six MLSA groups. Two MLSA groups contained reference strains of Rahnella genomospecies 2 and 3, three groups clustered within the Rahnella clade with no known type or reference strains and the last group contained the type strain of E. americana. DNA-DNA relatedness assays using both the microplate and fluorometric methods, confirmed that each of the five Rahnella MLSA groups formed separate taxa. Rahnella genomospecies 2 and 3 were previously not formally described due to a lack of distinguishing phenotypic characteristics. In the present study, all five Rahnella MLSA groups were phenotypically differentiated from each other and from R. aquatilis. Therefore we propose to classify the strains from symptomatic oak, alder and walnut and buprestid beetles as: Rahnella victoriana sp. nov. (type strain FRB 225(T)=LMG 27717(T)=DSM 27397(T)), Rahnella variigena sp. nov. (previously Rahnella genomosp. 2, type strain CIP 105588(T)=LMG 27711(T)), Rahnella inusitata sp. nov. (previously Rahnella genomosp. 3, type strain DSM 30078(T)=LMG 2640(T)), Rahnella bruchi sp. nov. (type strain FRB 226(T)=LMG 27718(T)=DSM 27398(T)) and Rahnella woolbedingensis sp. nov. (type strain FRB 227(T)=LMG 27719(T)=DSM 27399(T)).

Interactions of three soil bacteria species with phyllosilicate surfaces in hybrid silica gels.

To simulate iron consumption in soils, iron leaching from silicate minerals due to three heterotrophic bacterial strains and a chemical treatment was studied using hybrid silica gel (HSG) doped with two phyllosilicates, nontronite (NAu-2) or low-iron-content montmorillonite (SWy-2). HSG methodology, a novel way of separating bacteria cells from a colloidal mineral source, consisted in embedding colloidal mineral particles into an amorphous porous silica matrix using a classical sol-gel procedure. Pantoae agglomerans PA1 and Rahnella aquatilis RA1 were isolated from silicate-rich soils, that is, beech and wheat rhizospheres (Vosges, France); Burkholderia sp. G5 was selected from acidic and nutrient-poor podzol soils (Vosges, France). Fe release from clay minerals and production of bacterial metabolites, that is, low molecular weight organic acids (LMWOA) and siderophores, were monitored. Two LMWOA profiles were observed with major gluconate production (> 9000 µM) for Burkholderia sp. G5 and moderate production of lactate, acetate, propionate, formate, oxalate, citrate, and succinate (< 300 µM) for R. aquatilis RA1 and P. agglomerans PA1. HSG demonstrated its usefulness in revealing clay mineral-microorganisms interactions. The effect of bacterial exsudates was clearly separated from physical contact effect.

The spoilage characteristics of Brochothrix thermosphacta and two psychrotolerant Enterobacteriacae in vacuum packed lamb and the comparison between high and low pH cuts.

The spoilage potential of Brochothrix thermosphacta, Serratia proteamaculans and Rahnella aquatilis was investigated in vacuum packaged high (5.9 to 6.4) and low (5.4 to 5.8) pH lamb. Vacuum packaged fore shank (m. extensor carpi radialis) and striploins (m. longissimus dorsi) (n=306) inoculated with ~100 CFU of individual bacteria were stored for twelve weeks at temperatures -1.5, 0, 2 and 7°C. Spoilage characteristics and bacterial numbers were recorded and analysed in comparison to un-inoculated control samples. All three bacterial species were shown to grow in vacuum packaged lamb of pH values between 5.4 and 6.4, when stored at chilled temperatures (-1.5 to 7°C) for up to 84 days. B. thermosphacta and S. proteamaculans caused spoilage to the meat under these conditions whilst R. aquatilis spoiled high pH meat at 7°C. These results go against previous beef models stipulating that Brochothrix and Enterobacteriacae species cannot grow on or cause spoilage of low pH meat in the absence of oxygen.

Isolation and identification of phytate-degrading rhizobacteria with activity of improving growth of poplar and Masson pine.

A number of soil microorganisms can convert insoluble forms of phosphorus (P) to an accessible form to increase plant yields. Phytate is such a large kind of insoluble organic phosphorus that plants cannot absorb directly in soil, so the objectives of this study were to isolate, screen phytate-degrading rhizobacteria (PDRB), and to select potential microbial inocula that could increase the P uptake by plants. In this study, a total of 24 soil samples were collected from natural habitats of eight poplar and pine planting areas from the eastern to southern China. 17 PDRB strains were preliminarily screened from the rhizosphere soil of poplars and pines by the visible decolorization in the phytate selective medium. The highest ratio of the total diameter (colony + halo zone) to the colony diameter of the isolates was JZ-GX1, 3.85. Afterward, 17 PDRB strains were further determined for their abilities to degrade sodium phytate based on the amount of liberated inorganic P in liquid phytate specific medium. The results showed that the phytase ability of the three highest PDRB strains: JZ-GX1, JZ-DZ1 and JZ-ZJ1 were up to 2.58, 2.36 and 2.24 U/mL, respectively, much better than most of the bacteria reported in previous studies. In the soil-plant experiment, compared to CK, the best three strains of PDRB all could significantly promote growth of poplar and Masson pine under container growing. The three efficient PDRB strains were identified as follow: JZ-GX1, Rahnella aquatilis, both JZ-DZ1 and JZ-ZJ1 being autofluorescent, Pseudomonas fluorescens, by 16S rDNA gene sequencing technology, Biolog Identification System and biological characterization. The present study suggests that the three screened PDRB strains would have great potential application as biological fertilizers in the future.

[Screening, identification and phosphate-solubilizing characteristics of Rahnella sp. phosphate-solubilizing bacteria in calcareous soil].

Several strains of phosphate-solubilizing bacteria were isolated and screened from the crop rhizosphere of calcareous soil in Shanxi Province of China. After repeated isolation and purification, the strain W25 with strong phosphate-solubilizing activity was obtained, and identified as Rahnella sp., based on the morphological, physiological and biochemical properties and the analysis of 16S rRNA gene sequence. Further studies on the W25 showed that the maximum phosphate-solubilizing capability of the W25 on tricalium phosphate, aluminum phosphate and ferric phosphate reached 385.5, 110.4 and 216.6 mg x L(-1), respectively. In the liquid culture with aluminum phosphate and ferric phosphate, the solubilized phosphorous by the W25 was significantly negatively correlated with the liquid pH, with the correlation coefficient being 0.56 and 0.81, respectively. Among the carbon and nitrogen sources, glucose and ammonium nitrate were the optimum for the solubilization of tricalium phosphate by W25. The utilization of carbon source was in the order of glucose > lactose > sucrose > mannitose > starch, and that of nitrogen source was in the order of ammonium nitrate > ammonium chloride > ammonium sulfate > potassium nitrate > sodium nitrate. Different nitrogen sources had greater effects on the production of organic acids by W25. Formic acid and acetic acid would be produced when the nitrogen source was NH4+, oxalic acid and succinic acid would be produced when the nitrogen source was NO3(-), and citric acid would be extra produced when the ammonium nitrate was used as the nitrogen source.

Culturable bacterial microbiota of Plagiodera versicolora (L.) (Coleoptera: Chrysomelidae) and virulence of the isolated strains.

Plagiodera versicolora (Laicharting, 1781) (Coleoptera: Chrysomelidae) is an important forest pest which damages many trees such as willow, poplar, and hazelnut. In order to find new microbes that can be utilized as a possible microbial control agent against this pest, we investigated the culturable bacterial flora of it and tested the isolated bacteria against P. versicolora larvae and adults. We were able to isolate nine bacteria from larvae and adults. The isolates were characterized using a combination of morphological, biochemical, and physiological methods. Additionally, we sequenced the partial sequence of the 16S rRNA gene to verify conventional identification results. Based on characterization studies, the isolates were identified as Staphylococcus sp. Pv1, Rahnella sp. Pv2, Rahnella sp. Pv3, Rahnella sp. Pv4, Rahnella sp. Pv5, Pantoea agglomerans Pv6, Staphylococcus sp. Pv7, Micrococcus luteus Pv8, and Rahnella sp. Pv9. The highest insecticidal activity against larvae and adults was obtained from M. luteus Pv8 with 50 and 40 % mortalities within 10 days after treatment, respectively. Extracellular enzyme activity of the bacterial isolates such as amylase, proteinase, lipase, cellulose, and chitinase was also determined. Consequently, our results show that M. luteus Pv8 might be a good candidate as a possible microbial control agent against P. versicolora and were discussed with respect to biocontrol potential of the bacterial isolates.

Draft genome sequence of Rahnella aquatilis strain HX2, a plant growth-promoting rhizobacterium isolated from vineyard soil in Beijing, China.

Rahnella aquatilis strain HX2 is a plant growth-promoting, disease-suppressive rhizobacterium that was isolated from a vineyard soil in Beijing, China. Here, we report the genome sequence of this strain, which provides a valuable resource for future research examining the mechanisms of traits associated with plant growth promotion and biocontrol.

Complete genome sequence of Rahnella aquatilis CIP 78.65.

Rahnella aquatilis CIP 78.65 is a gammaproteobacterium isolated from a drinking water source in Lille, France. Here we report the complete genome sequence of Rahnella aquatilis CIP 78.65, the type strain of R. aquatilis.

[Chemical characterisation of structural components of Rahnella aquatilis lipopolysaccharides].

The studies of lipopolysaccharides (LPS) of eight Rahnella aquatilis strains, isolated from different sources have shown that they contain both S- and R-types of molecules. This fact is evidenced by the presence of high-molecular fraction of O-specific polysaccharides (O-PS) and low-molecular fraction of core oligosaccharides. The predominant monosaccharides of core oligosaccharides were glucose, galactose. The presence of only one high-molecular fraction O-PS was a characteristic feature of all investigated strains. The predominant monosaccharides of O-PS were galactose, glucose, mannose, rhamnose and fucose. 3-hydroxytetradecanoic (48.9-93.1%), dodecanoic (2.9-12.1%), tetradecanoic (4.1-25.3%) and hexadecanoic (2.8-15.3%) acids have been obtained in lipids A of LPS depending on the strain. The presence in lipid A of R. aquatilis of only 3-hydroxytetradecanoic acid which is characteristic of Enterobacteriaceae proves the correct ascribing of the isolated strains to this family.