Genera Acidovorax Research Articles

Nitrate Stimulates Anaerobic Microbial Arsenite Oxidation in Paddy Soils.

Arsenic (As) bioavailability to rice plants is elevated in flooded paddy soils due to reductive mobilization of arsenite [As(III)]. However, some microorganisms are able to mediate anaerobic As(III) oxidation by coupling to nitrate reduction, thus attenuating As mobility. In this study, we investigated the impact of nitrate additions on As species dynamics in the porewater of four As-contaminated paddy soils. The effects of nitrate on microbial community structure and the abundance and diversity of the As(III) oxidase (aioA) genes were quantified using 16S rRNA sequencing, quantitative PCR, and aioA gene clone libraries. Nitrate additions greatly stimulated anaerobic oxidation of As(III) to As(V) and decreased total soluble As in the porewater in flooded paddy soils. Nitrate additions significantly enhanced the abundance of aioA genes and changed the microbial community structure by increasing the relative abundance of the operational taxonomic units (OTUs) from the genera Acidovorax and Azoarcus. The aioA gene sequences from the Acidovorax related OTU were also stimulated by nitrate. A bacterial strain (ST3) belonging to Acidovorax was isolated from nitrate-amended paddy soil. The strain was able to oxidize As(III) and Fe(II) under anoxic conditions using nitrate as the electron acceptor. Abiotic experiments showed that Fe(II), but not As(III), could be oxidized by nitrite. These results show that nitrate additions can stimulate As(III) oxidation in flooded paddy soils by enhancing the population of anaerobic As(III) oxidizers, offering a potential strategy to decrease As mobility in As-contaminated paddy soils.

Extracellular Polyhydroxyalkanoate Depolymerase by Acidovorax sp. DP5.

Bacteria capable of degrading polyhydroxyalkanoates (PHA) by secreting extracellular depolymerase enzymes were isolated from water and soil samples collected from various environments in Malaysia. A total of 8 potential degraders exhibited clear zones on poly(3-hydroxybutyrate) [P(3HB)] based agar, indicating the presence of extracellular PHA depolymerase. Among the isolates, DP5 exhibited the largest clearing zone with a degradation index of 6.0. The highest degradation activity of P(3HB) was also observed with depolymerase enzyme of DP5 in mineral salt medium containing P(3HB). Based on biochemical characterization and 16S rRNA cloning and sequencing, isolate DP5 was found to belong to the genus Acidovorax and subsequently named as Acidovorax sp. DP5. The highest extracellular depolymerase enzyme activity was achieved when 0.25% (w/v) of P(3HB) and 1 g/L of urea were used as carbon and nitrogen source, respectively, in the culture media. The most suitable assay condition of the depolymerase enzyme in response to pH and temperature was tested. The depolymerase produced by strain Acidovorax sp. DP5 showed high percentage of degradation with P(3HB) films in an alkaline condition with pH 9 and at a temperature of 40°C.

The genetic toolbox for Acidovorax temperans

Environmental isolates belonging to the genus Acidovorax play a crucial role in degrading a wide range of pollutants. Studies on Acidovorax are currently limited for many species due to the lack of genetic tools. Here, we described the use of the replicon from a small, cryptic plasmid indigenous to Acidovorx temperans strain CB2, to generate stably maintained shuttle vectors. In addition, we have developed a scarless gene knockout technique, as well as establishing green fluorescent protein (GFP) reporter and complementation systems. Taken collectively, these tools will improve genetic manipulations in the genus Acidovorax.

Pseudacidovorax austerolens sp. nov., a nifH bacterium isolated from Himalayan valley soil, India

Gram-negative, rod-shaped bacterial strain ptl-2T, was isolated from soil of the Himalayan valley, India. The bacterial strain ptl-2T has been characterized using a polyphasic taxonomic approach including morphological characterization, fatty acid analysis, biochemical tests, 16S rRNA and nifH gene sequence analysis. 16S rRNA gene sequence analysis showed that the strain ptl-2T belonged to the genus Pseudacidovorax and is closely related to Pseudacidovorax intermedius (99.3 % similarity). It showed <97 % similarity to species of the genera Acidovorax, Alicycliphilus, Xylophilus, Giesbergeria and Simplicispira. The generic assignment has been confirmed on the basis of chemotaxonomic data, which revealed the fatty acid profile, characteristic of the genus Pseudacidovorax, consisting of C16:0 (21.78) and C18:1w7c (19.78) as major fatty acids. Phylogenetic, chemotaxonomic and phenotypic analysis based on signature sequences, DNA-DNA hybridization and physiological characterizations, confirms that strain ptl-2T represents a different species of the genus Pseudacidovorax for which the name Pseudacidovorax austerolens sp. nov. is proposed. The type strain ptl-2T (= CCUG 58759T, = DSM 24877T) has been submitted to two culture collection centres. GenBank accession numbers for the 16S rRNA and nifH sequence of strain Pseudacidovorax austerolens ptl-2T are FJ581042 and GQ249664, respectively.

Insights into Bacterial Community Structure of a Commercial Copper Bioheapleaching Plant: Growth of Heterotrophic Bacteria in the System

The distribution and diversity of bacterial community in Zijinshan commercial non-aeration copper bioheapleaching system operated at pH 0.8 for three years were investigated. The 24 meters high heap was cut off by mechanical digger. On the trapezoidal cross-section of the heap, 9 ore samples were taken from different vertical and horizontal locations and investigated by 16S rRNA gene clone library. Another 3 liquid samples from raffinate solution pond, spray solution pond and pregnant solution pond were also applied to 16S rRNA gene clone library analysis. The retrieved 1166 clone sequences from 12 samples were mainly related to genus Acidithiobacillus (42.36%), genus Leptospirillum (37.73%) and genus Sulfobacillus (6.52%). Relative high amount of heterotrophic bacteria were distributed at the ore surface in the internal part of the heap and in the liquid samples respectively. The retrieved heterotrophic bacterial sequences were mainly related to genus Acidiphilium (accounting 11.11% to 32.00% percent in the liquid samples), genus Acidovorax (accounting 12.37% in A1 sample), genus Pelomonas (accounting 4.17% to 10.31% in several ore samples) and genus Aquabacterium (accounting 10.31% in C2 sample). Bacterial diversity in the heap was increased from the surfcae layer to the interior of the heap. The proportion of genus Leptospirillum horizontally increased from the inner to the outer part while vertically decreased from lower depth (2-3 years leaching time) to higher depth(3-6 month leaching time), and reverse correlation of genus Acidithiobacillus was found in the heap. Our finding indicated that heterotrophic bacteria may play very important roles in the commercial bioheapleaching system, and revealed high distribution of genus Leptospirillum in the outer part of this non-aerated heap.

Isolation and functional analysis of denitrifiers in an aquifer with high potential for denitrification

Aquifers are among the main freshwater sources. The Raigón aquifer is susceptible to contamination, mainly by nitrate and pesticides, such as atrazine, due to increasing agricultural activities in the area. The capacity of indigenous bacteria to attenuate nitrate contamination in different wells of this aquifer was assessed by measuring denitrification rates with either acetate plus succinate or nitrate amendments. Denitrification activity in nitrate-amended assays was significantly higher than in unamended assays, particularly in groundwater from wells where nitrate concentration was 33.5mgL−1 or lower. Furthermore, groundwater denitrifiers capable of using acetate or succinate as electron donors were isolated, identified by 16S rRNA gene sequencing and evaluated for functional denitrification genes (nirS, nirK and nosZ). Phylogenetic affiliation of 54 isolates showed that all members belonged to nine different genera within the Proteobacteria (Bosea, Ochrobactrum, Azospira, Zoogloea, Acidovorax, Achromobacter, Vogesella, Stenotrophomonas and Pseudomonas). In addition, isolate AR28 that clustered separately from validly described species could potentially belong to a new genus. The majority of the isolates were related to species belonging to previously reported denitrifying genera. However, the phylogeny of the nirS and nosZ genes revealed new sequences of these functional genes. To our knowledge, this is the first isolation and sequencing of the nirS gene from the genus Vogesella, as well as the nosZ gene from the genera Acidovorax and Zoogloea. The results indicated that indigenous bacteria in the Raigón aquifer had the capacity to overcome high nitrate contamination and exhibited functional gene diversity.

Microbial communities along biogeochemical gradients in a hydrocarbon‐contaminated aquifer

Micro-organisms are known to degrade a wide range of toxic substances. How the environment shapes microbial communities in polluted ecosystems and thus influences degradation capabilities is not yet fully understood. In this study, we investigated microbial communities in a highly complex environment: the capillary fringe and subjacent sediments in a hydrocarbon-contaminated aquifer. Sixty sediment sections were analysed using terminal restriction fragment length polymorphism (T-RFLP) fingerprinting, cloning and sequencing of bacterial and archaeal 16S rRNA genes, complemented by chemical analyses of petroleum hydrocarbons, methane, oxygen and alternative terminal electron acceptors. Multivariate statistics revealed concentrations of contaminants and the position of the water table as significant factors shaping the microbial community composition. Micro-organisms with highest T-RFLP abundances were related to sulphate reducers belonging to the genus Desulfosporosinus, fermenting bacteria of the genera Sedimentibacter and Smithella, and aerobic hydrocarbon degraders of the genus Acidovorax. Furthermore, the acetoclastic methanogens Methanosaeta, and hydrogenotrophic methanogens Methanocella and Methanoregula were detected. Whereas sulphate and sulphate reducers prevail at the contamination source, the detection of methane, fermenting bacteria and methanogenic archaea further downstream points towards syntrophic hydrocarbon degradation.

Pyrosequencing Analysis of the Bacterial Community in Drinking Water Wells

Wells used for drinking water often have a large biomass and a high bacterial diversity. Current technologies are not always able to reduce the bacterial population, and the threat of pathogen proliferation in drinking water sources is omnipresent. The environmental conditions that shape the microbial communities in drinking water sources have to be elucidated, so that pathogen proliferation can be foreseen. In this work, the bacterial community in nine water wells of a groundwater aquifer in Northern Mexico were characterized and correlated to environmental characteristics that might control them. Although a large variation was observed between the water samples, temperature and iron concentration were the characteristics that affected the bacterial community structure and composition in groundwater wells. Small increases in the concentration of iron in water modified the bacterial communities and promoted the growth of the iron-oxidizing bacteria Acidovorax. The abundance of the genera Flavobacterium and Duganella was correlated positively with temperature and the Acidobacteria Gp4 and Gp1, and the genus Acidovorax with iron concentrations in the well water. Large percentages of Flavobacterium and Pseudomonas bacteria were found, and this is of special concern as bacteria belonging to both genera are often biofilm developers, where pathogens survival increases.

Description of Acidovorax wautersii sp. nov. to accommodate clinical isolates and an environmental isolate, most closely related to Acidovorax avenae

Three Gram-negative strains, NF 1078(T), NF 1598 and NF 1715, were isolated from clinical (two) and environmental (one) samples, respectively. Sequence analysis of the 16S rRNA genes revealed similarity of 100% among the three strains and next highest similarity to the type strain of Acidovorax avenae (98.16%). The three strains were able to acidify lactose and rhamnose on low peptone phenol red agar and to alkalinize citrate on Simmons' agar and were negative for nitrate reduction. The DNA G+C content of strain NF 1078(T) was 67.1 mol%. The level of DNA-DNA relatedness between this strain and the type strains of A. avenae (ATCC 19860(T), LMG 2117(T)) was 29%. Based on these phylogenetic, phenotypic and genotypic analyses, the three strains could be distinguished clearly from all other recognized Acidovorax species and should be classified as representatives of a novel species of the genus Acidovorax, for which the name Acidovorax wautersii sp. nov. is proposed. The type strain is NF 1078(T) (=LMG 26971(T)=CCUG 62584(T)).

Acidovorax citrulli: generating basic and applied knowledge to tackle a global threat to the cucurbit industry

Acidovorax citrulli is the causal agent of bacterial fruit blotch (BFB) of cucurbit plants. In recent years, the disease has spread to many parts of the world, mainly via the inadvertent distribution of contaminated commercial seeds. Because of the costly lawsuits filed by growers against seed companies and the lack of efficient management methods, BFB represents a serious threat to the cucurbit industry, and primarily to watermelons and melons. Despite the economic importance of the disease, little is known about the basic aspects of A. citrulli pathogenesis. Nevertheless, the release of the genome of one A. citrulli strain, as well as the optimization of molecular manipulation and inoculation methods, has prompted basic studies and allowed advances towards an understanding of A. citrulli pathogenicity. In this article, we summarize the current knowledge about this important pathogen, with emphasis on its epidemiology and the factors involved in its pathogenicity and virulence. Bacteria; Betaproteobacteria; order Burkholderiales; family C omamonadaceae; genus Acidovorax; species citrulli. Gram-negative, strictly aerobic, rod-shaped; average dimensions of 0.5 μm × 1.7 μm; motile by means of an ~5.0-μm-long polar flagellum; colonies on King's medium B are round, smooth, transparent and nonpigmented; optimal temperatures for growth around 27-30 °C; induces a hypersensitive response on nonhost tobacco and tomato leaves. Acidovorax citrulli strains are pathogenic to various species of the Cucurbitaceae family, including watermelon, melon, squash, pumpkin and cucumber. Significant economic losses have been reported in watermelon and melon. Watermelon and melon seedlings and fruits are highly susceptible to A. citrulli. Typical seedling symptoms include water-soaked lesions on cotyledons that are often adjacent to the veins and later become necrotic, lesions on the hypocotyl, and seedling collapse and death. On watermelon fruits, symptoms begin as small, irregular, water-soaked lesions which later extend through the rind, turn brown and crack. On melon fruits, symptoms are characterized by small, often sunken rind lesions and internal fruit decay. Symptoms on the leaves of mature plants are difficult to diagnose because they are often inconspicuous or similar to those caused by other biotic or abiotic stresses. When they occur, leaf lesions can spread along the midrib and main veins. Lesions appear dark-brown to black on watermelon and light to reddish-brown on melon. Bacterial fruit blotch of cucurbits at APSnet, http://www.apsnet.org/edcenter/intropp/lessons/prokaryotes/Pages/BacterialBlotch.aspx; bacterial fruit blotch guide from ASTA, http://www.amseed.com/pdfs/DiseaseGuide-BFB-English.pdf; Acidovorax citrulli AAC00-1 genome at JGI, http://genome.jgi-psf.org/aciav/aciav.info.html.

Acidovorax radicis sp. nov., a wheat-root-colonizing bacterium

Strain N35(T) was isolated from surface-sterilized wheat roots and is a Gram-negative, aerobic, motile straight rod. Strain N35(T) tested oxidase-positive and catalase-negative and grew optimally at pH 7.0, 30 °C and in the absence of NaCl. 16S rRNA gene sequence analysis showed over 97 % sequence similarity to strains of the environmental species Acidovorax delafieldii, A. facilis, A. defluvii, A. temperans, A. caeni and A. soli, as well as Acidovorax valerianellae, A. anthurii and Simplicispira metamorpha. DNA-DNA hybridization between strain N35(T) and phylogenetically closely related type strains was 25.3-55.7 %, which clearly separates the strain from these closely related species. Additionally, phenotypic properties, such as substrate metabolism profiles as determined by a Biolog GN2 assay and cell-wall fatty acid profiles, particularly contents of the fatty acids C(16 : 0), C(16 : 1)ω7c/t, C(17 : 0), C(17 : 0) cyclo, C(18 : 0) cyclo and C(19 : 0) cyclo, facilitated the differentiation of the newly isolated strain N35(T) from its closest relatives. The isolate underwent phenotypic variation at high frequency in laboratory media. The DNA G+C content was 64.9 mol%. We propose that strain N35(T) is classified as a representative of a novel species within the genus Acidovorax, and suggest the name Acidovorax radicis sp. nov. The type strain is strain N35(T) ( = DSM 23535(T) = LMG 25767(T)).

Bacteria from drinking water supply and their fate in gastrointestinal tracts of germ-free mice: A phylogenetic comparison study

Microorganisms in drinking water sources may colonize in gastrointestinal (GI) tracts and this phenomenon may pose a potential health risk especially to immunocompromised population. The microbial community diversity of the drinking water was compared with the GI tracts of the mice using phylogenetic and statistical analyses of 16S rRNA gene sequences. A group of germ-free mice were fed with drinking water from public water supply that passed through an automated watering system with documented biofilm accumulation. From drinking water and GI tracts of the germ-free mice, 179 bacteria were isolated and 75 unique 16S rRNA gene phylotypes were sequenced as operational taxonomic unit (OTU, >97% similarity). Three major groups of the genus Acidovorax (21%), Variovorax (42%) and Sphingopyxis (15%) were found in drinking water. Three major groups of the genus Ralstonia (24%), Staphylococcus (20%) and Bosea (22%) were found in GI tracts. Ralstonia (6%, 24%), Sphingopyxis (15%, 2%), Bacillus (3%, 5%), Escherichia coli (3%, 2%) and Mesorhizobium (3%, 5%) were found in both sources – drinking water and GI tract. A lineage-per-time plot shows that the both bacterial communities have convex shape lines, suggesting an excess of closely related ecotypes. A significant F ST test (0.00000–0.00901) coupled with an insignificant P test (0.07–0.46) implies that the tree contained several clades of closely related bacteria. Both phylogenetic and statistical results suggest a correlation between the bacterial communities originating in the drinking water and those associated with the GI tracts. The GI tract showed a higher genetic diversity than the drinking water, but a similar lineage-per-time plot was obtained overall. It means a sudden evolutionary transformation and colonization occurred with high selective forces.

Acidovorax soli sp. nov., isolated from landfill soil

A Gram-negative, aerobic, rod-shaped, non-motile strain, BL21(T), was isolated from landfill soil in Pohang, Korea. Strain BL21(T) grew optimally at pH&emsp14;7.0, 30 °C and 0 % NaCl (w/v). Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain BL21(T) belonged to the class Betaproteobacteria and was related to the genus Acidovorax. The 16S rRNA gene sequence of strain BL21(T) was less than 98.30 % similar to those of other species in the genus Acidovorax. DNA-DNA hybridization values with phylogenetically related species of the genus Acidovorax were only 11.7-28.4 %. The major fatty acid components included summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)ω7c), C(16 : 0), C(18 : 1)ω7c and C(10 : 0) 3-OH. The DNA G+C content was 60.9 mol%. For these reasons, strain BL21(T) (=KCTC 22399(T) =JCM 15909(T)) is proposed as a novel species in the genus Acidovorax, with the name Acidovorax soli sp. nov.

Verminephrobacter eiseniae gen. nov., sp. nov., a nephridial symbiont of the earthworm Eisenia foetida (Savigny)

A Gram-negative, flagellated, heterotrophic, catalase-negative, rod-shaped bacterium previously identified as an earthworm symbiont was isolated from nephridia of the earthworm Eisenia foetida. Comparisons of 16S rRNA gene sequences indicated its relatedness to the betaproteobacterial genus Acidovorax and the novel isolates shared 92-94% sequence similarity with recognized species of this genus. Gene sequence phylogenies revealed that the group of earthworm symbionts formed a cohesive and independent clade. The DNA G+C content was 67.0+/-0.2 mol%. Major fatty acids were C(16:0), C(16:1)omega7c and C(17:0) cyclo. While capable of growing in fully aerated media, all isolates favoured low oxygen concentrations and all required biotin or a mix of amino acids in order to grow on defined mineral media. Based on phylogenies inferred from three housekeeping gene sequences (gap, recA and rpoC), DNA-DNA hybridization values, the unique ecology and the distinct physiology of the novel strains, the new genus Verminephrobacter gen. nov. is proposed for the earthworm nephridial symbionts. The name Verminephrobacter eiseniae sp. nov. is proposed for the type species with strain EF01-2(T) (=ATCC BAA-1489(T)=DSM 19286(T)) as the type strain of the type species.

Investigations on influencing plant-associated bacteria in tissue cultures of black locust (Robinia pseudoacacia L.)

During tissue culture of black locust (Robinia pseudoacacia L.), serious problems with plant-associated bacteria led to a reduction of propagation potential in several clones. Four dominant strains of plant-associated bacteria could be isolated and were assigned to the genera Acidovorax, Dyella, Microbacterium and Sphingomonas. Out of five essential oils tested, thyme and lemongrass oil at a concentration of 0.03% each and 0.015% of both oils in combination clearly inhibited the growth of these bacteria strains on bacteriologic medium. There were no significant differences in total bacterial population density when penicillin, thyme and lemongrass oil or thyme plus lemongrass oil were added to the plant propagation media. The use of lemongrass oil changed the proportion of dominant bacterial strains.

Acidovorax caeni sp. nov., a denitrifying species with genetically diverse isolates from activated sludge

Four Gram-negative, rod-shaped, non-spore-forming, denitrifying isolates were obtained from the activated sludge of an aerobic-anaerobic wastewater treatment plant in Belgium. Analysis of repetitive sequence-based PCR showed that the four isolates were genetically different from each other. Results of 16S rRNA gene sequence analysis and DNA-DNA hybridization experiments indicated that these four isolates were affiliated to the genus Acidovorax and could be differentiated from all recognized species of the genus. Analysis of whole-cell proteins and results of physiological and biochemical tests allowed differentiation of the new isolates from their closest phylogenetic neighbours. These new isolates therefore represent a novel species of the genus Acidovorax, for which the name Acidovorax caeni sp. nov. is proposed. The type strain is R-24608(T) (=LMG 24103(T) =DSM 19327(T)).

Diversity of Ferrous Iron-Oxidizing, Nitrate-Reducing Bacteria and their Involvement in Oxygen-Independent Iron Cycling

In previous studies, three different strains (BrG1, BrG2, and BrG3) of ferrous iron-oxidizing, nitrate-reducing bacteria were obtained from freshwater sediments. All three strains were facultative anaerobes and utilized a variety of organic substrates and molecular hydrogen with nitrate as electron acceptor. In this study, analyses of 16S rDNA sequences showed that strain BrG1 was affiliated with the genus Acidovorax, strain BrG2 with the genus Aquabacterium, and strain BrG3 with the genus Thermomonas. Previously, bacteria similar to these three strains were detected with molecular techniques in MPN dilution series for ferrous iron-oxidizing, nitrate-reducing bacteria inoculated with different freshwater sediment samples. In the present study, further molecular analyses of these MPN cultures indicated that the ability to oxidize ferrous iron with nitrate is widespread amongst the Proteobacteria and may also be found among the Gram-positive bacteria with high GC content of DNA. Nitrate-reducing bacteria oxidized ferrous iron to poorly crystallized ferrihydrite that was suitable as an electron acceptor for ferric iron-reducing bacteria. Biologically produced ferrihydrite and synthetically produced ferrihydrite were both well suited as electron acceptors in MPN dilution cultures. Repeated anaerobic cycling of iron was shown in a coculture of ferrous iron-oxidizing bacteria and the ferric iron-reducing bacterium Geobacter bremensis. The results indicate that iron can be cycled between its oxidation states +II and +III by microbial activities in anoxic sediments.

Acidovorax-like symbionts in the nephridia of earthworms.

Dense accumulations of bacteria in the excretory organs, nephridia, were first described more than 75 years ago in members of the annelid family Lumbricidae (earthworms). These nephridial symbionts were assumed to play a role in the degradation of proteins in the excretory fluid for nitrogen recycling. In the present study, the phylogenetic affiliation of the nephridial bacteria of the earthworms Lumbricus terrestris, Aporrectodea tuberculata, Octolasion lacteum and Eisenia foetida was resolved. The 16S rRNA gene sequences of the symbionts formed a monophyletic cluster within the genus Acidovorax. Similarity between symbiont sequences from different host species was 95.5-97.6%, whereas similarity was> 99% between symbiont sequences from individuals of the same species. Densely packed bacteria were detected in the ampulla of the nephridia by fluorescence in situ hybridization (FISH) using Acidovorax-specific oligonucleotide probes. No other bacterial cells could be found by FISH, although a few sequences other than Acidovorax had been found by PCR and cloning. These results suggest that the Acidovorax-earthworm symbiosis is a stable, host-specific association that has evolved from a common bacterial ancestor. Given the close phylogenetic relationship of the symbionts to proteolytic, free-living Acidovorax species, they may indeed play a role in protein degradation during nitrogen excretion by earthworms.

Acidovorax valerianellae sp. nov., a novel pathogen of lamb's lettuce [Valerianella locusta (L.) Laterr

Bacterial spot disease of lamb's lettuce [Valerianella locusta (L.) Laterr.] was first observed in fields in 1991. This new bacterial disease is localized in western France in high-technology field production of lamb's lettuce for the preparation of ready-to-use salad. Nineteen strains isolated in 1992 and 1993 from typical black leaf spots of naturally infected lamb's lettuce were characterized and compared with reference strains of Acidovorax and Delftia. The pathogenicity of the 19 strains was confirmed by artificial inoculation. Biochemical and physiological tests, fatty acid profiles, DNA-DNA hybridization and other nucleic acid-based tests were performed. A numerical taxonomic analysis of the 19 lamb's lettuce strains showed a single homogeneous phenon closely related to previously described phytopathogenic taxa of the genus Acidovorax. DNA-DNA hybridization studies showed that the lamb's lettuce strains were 91-100% related to a representative strain, strain CFBP 4730(T), and constituted a discrete DNA hybridization group, indicating that they belong to the same novel species. Results from DNA-rRNA hybridization, 16S rRNA sequence analysis and fatty acid analysis studies confirmed that this novel species belongs to the beta-subclass of the Proteobacteria and, more specifically, to the family Comamonadaceae and the genus Acidovorax. The name Acidovorax valerianellae sp. nov. is proposed for this novel taxon of phytopathogenic bacteria. The type strain is strain CFBP 4730(T) (= NCPPB 4283(T)).

Polyphasic Characterization of Poly-3-hydroxybutyrate-co-3-hydroxyvalerate (P(HB-co-HV)) Metabolizing and Denitrifying Acidovorax sp. Strains

For the purpose of denitrification in small drinking water plants, a bacterial mixed population was isolated from a packed bed column bioreactor with poly-3-hydroxybutyrate-co-3-hydroxyvalerate (P(HB-co-HV)) as a substrate for the denitrification of ground water (10 degrees C). Isolates 2nIII from the mixed culture, with the ability to denitrify and metabolize P(HB-co-HV), were used as starter cultures for the elimination of nitrate in ground water. The strains were characterized by diverse techniques. Classical phenotypic studies lead to rRNA group III of the genus Pseudomonas. Results obtained by molecular techniques demonstrated that the 2nIII strains are members of the Comamonadaceae and shows similarities to the genus Acidovorax. However, an integration of the 2nIII isolates within one of the known Acidovorax species is not possible for the moment. The 2nIII starter cultures clustered close to Av. temperans according to their whole cell proteins and fatty acids, whereas in DNA/DNA hybridization no significant DNA binding (< 25%) was found. In contrast a significant but low degree of DNA/DNA hybridization was found between the 2nIII strains and Av. facilis and Av. delafieldii. Our polyphasic results lead to the conclusion that the 2nIII strains may constitute a separate Acicdovorax species.