Effect of Environmental Factors in Aerobiosis and Anaerobiosis on the Growth and Activity of Propionic Acid Bacteria Evaluated by Pressure Measurement

The effects of air, N₂ and CO₂ atmospheres on the growth and metabolism of P. shermanii CIP 10 30 27 under “optimum conditions” (pH 6.5, NaCl 0 %, temperature 30 °C) and “stressful conditions” (pH 5.3, NaCl 2.1 %, temperature 20 °C) simulating cheese ripening conditions were investigated using a pressure measurement technique. Under both conditions, the inhibition of growth was at its maximum when a CO₂ atmosphere with an increase in the lag phase (L_P) from 3 to 80 h and a reduction in the maximum rate of pressure variation (V_max,P) from 2.5 to 0.4 kPa/h was used. The production of metabolites was not affected under “optimum conditions”, but under “stressful conditions”, propionate production increased and higher ratios of propionate to acetate of up to 2.45 were observed. The effects of air and N₂ atmospheres on both growth and metabolite production were also examined.     

NADP‐glutamate dehydrogenase activity in nonstarter lactic acid bacteria: effects of temperature,pH and NaCl on enzyme activity and expression

Aims: To screen the glutamate dehydrogenase (GDH) activity of nonstarter lactic acid bacteria (NSLAB) and to determine the effects of temperature, pH and NaCl values used for cheese ripening on enzyme activity and expression of GDH gene. Methods and Results: A subcellular fractionation protocol and specific enzyme assays were used. The effect of temperature, pH and NaCl on enzyme activity was evaluated. The expression of GDH gene was monitored by real‐time PCR. One selected strain was also used as adjunct starter for cheese making to evaluate the catabolism of free amino acids and the production of volatile organic compounds (VOC) during cheese ripening. The cytoplasm fraction of all strains showed in vitro NADP‐dependent GDH activity. NADP‐GDH activity was markedly strain dependent and varied according to the interactions between temperature, pH and NaCl. Lactobacillus plantarum DPPMA49 showed the highest NADP‐GDH activity under temperature, pH and NaCl values found during cheese ripening. RT‐PCR analysis revealed that GDH expression of Lact. plantarum DPPMA49 was down‐expressed by low temperature (<13°C) and over‐expressed by NaCl (1·87–5·62%). According to NADP‐GDH activity, the highest level of VOC (alcohols, aldehydes, miscellaneous and carboxylic acids) was found in cheeses made with DPPMA49. Conclusions: The results of this study may be considered as an example of the influence of temperature, pH and NaCl on enzyme activity and expression of functional genes, such as GDH, in cheese‐related bacteria. Significance and Impact of the Study: It focuses on the phenotypic and molecular characterization of the NADP‐GDH in lactobacilli under cheese‐ripening conditions. The findings of this study contribute to the knowledge about enzymes involved in the catabolism of amino acids, to be used as an important selection trait for cheese strains.     

Growth Conditions Influence Expression of Cell Surface Hydrophobicity of Staphylococci and Other Wound Infection Pathogens

The initial adhesion of microbes to tissue and solid surfaces can be mediated by hydrophobic interaction. Expression of microbial cell surface hydrophobicity (CSH) is influenced by growth conditions, and often best expressed after growth under nutrient-poor conditions, or “starvation.” In the present study, the CSH of 133 strains of Enterobacteriaceae, Staphylococcus aureus, coagulase-negative staphylococci, Enterococcus faecalis, group A streptococcus, Pseudomonas aeruginosa, Clostridium perfringens, Bacteroides fragilis, Peptococcus magnus, and of 8 Candida albicans strains was measured by the salt aggregation test after growth on hematin agar in a 5% CO₂ atmosphere, or under anaerobiosis. Cells of all but 8 strains expressed pronounced or moderate CSH, i.e., they aggregated in 0.01-2 M ammonium sulfate. When the agar surface was covered by human serum (diluted 1:5) to mimic growth conditions in a wound, 94 strains expressed higher CSH, and 44 strains the same CSH as after growth without serum. The CSH of 12 strains of different species was measured after growth on blood, hematin and PDM agar, with or without serum, and in an aerobic or a 5% CO₂ atmosphere. The highest CSH was expressed after growth in 5% CO₂ with serum, and the lowest growth after on blood agar in aerobic atmosphere. Identical results were obtained with native and heat-inactivated (56 C, 20 min) serum. The reduced surface tension obtained in 5% CO₂, as well as yet unidentified serum factors, promotes expression of CSH.     

Stress Tolerance and Genetic Variability of Phosphate-solubilizing Fluorescent Pseudomonas from the Cold Deserts of the Trans-Himalayas

Nineteen efficient phosphate-solubilizing fluorescent Pseudomonas from the cold deserts of the trans-Himalayas were screened for stress tolerance against temperature, alkalinity, salinity, calcium salts, and desiccation. Phylogenetic analysis based on 16S rRNA gene sequencing placed these bacteria under three groups with fourteen strains in Group I including Pseudomonas trivialis and P. poae, two strains in Group II together with Pseudomonas kilonensis and P. corrugata, and three strains in Group III along with Pseudomonas jessenii and P. moraviensis. Genetic diversity assessed by ERIC and BOX-PCR revealed variability among strains belonging to the same phylogenetic groups. Cluster analysis based on the growth characteristics under regimes of different stress levels placed the strains into three distinct clusters displaying no correlation to their phylogenetic groups. Stress-tolerant strains differed in the level of decline in phosphate solubilization under increasing intensity of various stress parameters. The highest decrease occurred with 5% CaCO_3, followed by 2.5% CaCO₃, pH 11, 5% NaCl, temperature of 37°C, 40% PEG, 5% CaSO₄, 2.5% NaCl, 2.5% CaSO₄, pH 9 and temperature of 15°C. Two strains belonging to Phylogenetic Group I exhibited higher phosphate solubilization at lower temperature. The results revealed that stress-tolerance ability was not limited to any particular phylogenetic group. Knowledge about the genetic variants of phosphate-solubilizing fluorescent Pseudomonas with potential for tolerance to desiccation, alkalinity, temperature, and salinity could be useful in understanding their ecological role under stressful environments of low phosphate availability.     

Dietary Restraint and Control Over “Wanting” Following Consumption of “Forbidden” Food

Eating behavior can be influenced by the rewarding value of food, i.e., “liking” and “wanting.” The objective of this study was to assess in normal‐weight dietary restrained (NR) vs. unrestrained (NU) eaters how rewarding value of food is affected by satiety, and by eating a nonhealthy perceived, dessert‐specific food vs. a healthy perceived, neutral food (chocolate mousse vs. cottage cheese). Subjects (24NR age = 25.0 ± 8.2 years, BMI = 22.3 ± 2.1 kg/m²; 26NU age = 24.8 ± 8.0 years, BMI = 22.1 ± 1.7 kg/m²) came to the university twice, fasted (randomized crossover design). Per test‐session “liking” and “wanting” for 72 items divided in six categories (bread, filling, drinks, dessert, sweets, stationery (placebo)) was measured, before and after consumption of chocolate mousse/cottage cheese, matched for energy content (5.6 kJ/g) and individual daily energy requirements (10%). Chocolate mousse was liked more than cottage cheese (P < 0.05). After consumption of chocolate mousse or cottage cheese, appetite and “liking” vs. placebo were decreased in NR and NU (P < 0.03), whereas “wanting” was only decreased in NR vs. NU (P ≤ 0.01). In NR vs. NU “wanting” was specifically decreased after chocolate mousse vs. cottage cheese; this decrease concerned especially “wanting” for bread and filling (P < 0.05). To conclude, despite similar decreases in appetite and “liking” after a meal in NR and NU, NR decrease “wanting” in contrast to NU. NR decrease “wanting” specifically for a nonhealthy perceived, “delicious,” dessert‐specific food vs. a nutritional identical, yet healthy perceived, slightly less “delicious,” “neutral” food. A healthy perceived food may thus impose greater risk for control of energy intake in NR.     

Influences of elevated CO2 and pest damage on the allocation of plant defense compounds in Bt‐transgenic cotton and enzymatic activity of cotton aphid

Abstract Plant allocation to defensive compounds by elevated CO₂‐grown non‐transgenic and transgenic Bt cotton in response to infestation by cotton aphid, Aphis gossypii (Glover) in open‐top chambers under elevated CO₂ were studied. The results showed that significantly lower foliar nitrogen concentration and Bt toxin protein occurred in transgenic Bt cotton with and without cotton aphid infestation under elevated CO_2. However, significantly higher carbon/nitrogen ratio, condensed tannin and gossypol were observed in transgenic Bt cotton “GK‐12” and non‐transgenic Bt cotton ‘Simian‐3’ under elevated CO_2. The CO₂ level and cotton variety significantly influenced the foliar nitrogen, condensed tannin and gossypol concentrations in the plant leaves after feeding by A. gossypii. The interaction between CO₂ level × infestation time (24 h, 48 h and 72 h) showed a significant increase in cotton condensed tannin concentrations, while the interaction between CO₂ level × cotton variety significantly decreased the true choline esterase (TChE) concentration in the body of A. gossypi. This study exemplified the complexities of predicting how transgenic and non‐transgenic plants will allocate defensive compounds in response to herbivorous insects under differing climatic conditions. Plant defensive compound allocation patterns and aphid enzyme changes observed in this study appear to be broadly applicable across a range of plant and herbivorous insect interactions as CO₂ atmosphere rises.     

Effect of several MAP compositions on the microbiological and sensory properties of Graviera cheese

The shelf life of Graviera cheese, a full fat cheese produced in Heraklion (Crete Greece), was investigated. Graviera cheese was stored at 4 °C for up to 90 days in polyamide packages under three different modified atmosphere compositions. Control cheeses were packaged in air whereas MAP mixtures were MAP₁: 40% CO₂/55% N₂/5% O₂, MAP₂: 60% CO₂/40% N₂ and MAP₃: 50% CO₂/50% N₂. Sampling of product was carried out every 10 days to investigate its sensory quality and microbiological characteristics. Ten trained panelists participated in the sensory panel to evaluate the cheeses for external appearance (color, texture), taste, and flavor in a scale from 1 to 10 (1 very poor, 10 very good). The microbiological analysis revealed that there were no colonies of Staphylococcus aureus and Listeria monocytogenes whereas both Escherichia coli and Total Viable Counts (TVC) increased strongly in control samples but were inhibited under all MAP compositions.     

Role of halotolerant phosphate‐solubilising bacteria on growth promotion of peanut (Arachis hypogaea) under saline soil

Soil salinity is a major abiotic stress that limits plant growth, and inoculating plant growth‐promoting rhizobacteria is a well‐known strategy to reduce stressors under adverse soil conditions. This study was conducted to assess the effect of halotolerant phosphate‐solubilising bacteria (PSB) on protecting peanut against salt stress. Four candidate strains: Bacillus megaterium (YM13), Enterobacter sp. (YM14), Providencia rettgeri (TPM23) and Ensifer adhaeren (TPMX5) showed strong tolerance to NaCl and high phosphate‐solubilising ability even at a NaCl concentration of 1.4 M. In addition, all four strains demonstrated variable levels of phosphate solubilisation activity in the presence of various carbon and nitrogen sources, indicating high phosphate‐solubilising efficacy. Germination and radicle length of peanut seedlings increased with inoculation of PSB under both control and saline conditions. Statistically significant increase in the root length (range: 25.71–49%), stem length (19–48%), number of leaves (12.5–37%) and root/shoot biomass were observed. This could be attributed to plant hormones (i.e., indole acetic acid [IAA], abscisic acid [ABA] and gibberellic acid [GA3]) and successful root colonisation by bacterial inoculants. Root colonisation was positively correlated to plant growth and shown to be influenced by soil conditions. In addition, the PSB also improved the levels of available P in soil. The most pronounced beneficial effect on the growth of peanut plants and soil available P content was observed in the inoculation of the PSB isolates with Ca₃(PO₄)₂ addition. This is the first report that describes Providencia rettgeri as a plant growth‐promoting bacterium that may be utilised to alleviate the negative effects of salt stress on peanut plants. This bacterial species may thus be potentially used as a biofertilizer for sustaining the growth of peanut in salt‐stressed soil and in mitigating soil stress conditions.     

Metabolic and proteomic adaptation of Lactobacillus rhamnosus strains during growth under cheese‐like environmental conditions compared to de Man,Rogosa, and Sharpe medium

The aim of this study was to demonstrate the metabolic and proteomic adaptation of Lactobacillus rhamnosus strains, which were isolated at different stages of Parmigiano Reggiano cheese ripening. Compared to de Man, Rogosa, and Sharpe (MRS) broth, cultivation under cheese‐like conditions (cheese broth, CB) increased the number of free amino acids used as carbon sources. Compared with growth on MRS or pasteurized and microfiltrated milk, all strains cultivated in CB showed a low synthesis of d,l ‐lactic acid and elevated levels of acetic acid. The proteomic maps of the five representative strains, showing different metabolic traits, were comparatively determined after growth on MRS and CB media. The amount of intracellular and cell‐associated proteins was affected by culture conditions and diversity between strains, depending on their time of isolation. Protein spots showing decreased (62 spots) or increased (59 spot) amounts during growth on CB were identified using MALDI‐TOF‐MS/MS or LC‐nano‐ESI‐MS/MS. Compared with cultivation on MRS broth, the L. rhamnosus strains cultivated under cheese‐like conditions had modified amounts of some proteins responsible for protein biosynthesis, nucleotide, and carbohydrate metabolisms, the glycolysis pathway, proteolytic activity, cell wall, and exopolysaccharide biosynthesis, cell regulation, amino acid, and citrate metabolism, oxidation/reduction processes, and stress responses.     

Possible nonanthropogenic origin of two methanogenic isolates from oil‐producing wells in the san miguelito field,ventura county,California

Two novel strains of rod‐shaped methanogens were isolated from oil‐producing wells of high temperature and moderate salinity in the San Miguelito field. The strains grow at 65°C in media containing 6% NaCl, form large aggregates of cell materials in liquid culture, and produce methane from H₂ and CO₂ only. The geochemistry and microbiology of the oil reservoir and surrounding areas support the view that not all methanogens isolated from oil wells are results of colonization during or after human exploration, and that methanogens can exist in situ under thermal, haline conditions.     

Overcoming Charge Collection Limitation at Solid/Liquid Interface by a Controllable Crystal Deficient Overlayer

Bulk and surface charge recombination of photoelectrode are two key processes that significantly hinder solar‐to‐fuel conversion of photoelectrochemical cell (PEC). In this study, the function of a “crystal‐deficient” overlayer is unveiled, which outperforms a traditionally used amorphous or crystalline overlayer in PEC water splitting by exhibiting a high conductivity and large electron diffusion length to enable unlimited electron collection. The optimized ≈2.5 nm thickness of the “crystal‐deficient” shell results in a depletion layer with a width of 3 nm, which overcomes the flat band limitation of the photovoltage and increases the light absorptivity in the wavelength range from 300 to 420 nm. In addition, a 50‐fold increase in the conductivity yields a one‐order‐of‐magnitude increase in the diffusion length of an electron (L_n )(≈20 μm), allowing for unlimited electron collection in the 1.9 μm TiO₂ nanowire array with the “crystal‐deficient” shell. The controllable “crystal‐deficient” overlayer in rutile TiO₂ nanowires photoanode achieves a photocurrent density greater than 2.0 mA cm^?2 at 1.23 V versus reversible hydrogen electrode (RHE), a 1.18% applied bias photon‐to‐current efficiency at 0.49 V versus RHE, a faradaic efficiency greater than 93.5% at 0.6 V versus Pt under air mass 1.5G simulated solar light illumination (100 mW cm^?2).     

Adaptation of Listeria monocytogenes in a simulated cheese medium: effects on virulence using the Galleria mellonella infection model

The aim of this study was to evaluate the effect of the acid and salt adaptation in a cheese‐based medium on the virulence potential of Listeria monocytogenes strains isolated from cheese and dairy processing environment using the Galleria mellonella model. Four L. monocytogenes strains were exposed to a cheese‐based medium in conditions of induction of an acid tolerance response and osmotolerance response (pH 5·5 and 3·5% w/v NaCl) and injected in G. mellonella insects. The survival of insects and the L. monocytogenes growth kinetics in insects were evaluated. The gene expression of hly, actA and inlA genes was determined by real‐time PCR. The adapted cells of two dairy strains showed reduced insect mortality (P < 0·05) in comparison with nonadapted cells. Listeria monocytogenes Scott A was the least virulent, whereas the cheese isolate C882 caused the highest insect mortality, and no differences (P > 0·05) was found between adapted and nonadapted cells. The gene expression results evidenced an overexpression of virulence genes in cheese‐based medium, but not in simulated insect‐induced conditions. Our results suggest that adaptation to low pH and salt in a cheese‐based medium can affect the virulence of L. monocytogenes, but this effect is strain dependent.

Significance and Impact of the Study

In this study, the impact of adaptation to low pH and salt in a cheese‐based medium on L. monocytogenes virulence was tested using the Wax Moth G. mellonella model. This model allowed the differentiation of the virulence potential between the L. monocytogenes strains. The effect of adaptation on virulence is strain dependent. The G. mellonella model revealed to be a prompt method to test food‐related factors on L. monocytogenes virulence.     

Soprano singing in gibbons

Diversifications in primate vocalization, including human speech, are believed to reflect evolutionary modifications in vocal anatomy and physiology. Gibbon song is acoustically unique, comprising loud, melodious, penetrating pure tone‐like calls. In a white‐handed gibbon, Hylobates lar, the fundamental frequency (f₀) of song sounds is amplified distinctively from the higher harmonics in normal air. In a helium‐enriched atmosphere, f₀ does not shift, but it is significantly suppressed and 2f₀ is emphasized. This implies that the source is independent of the resonance filter of the supralaryngeal vocal tract (SVT) in gibbons, in contrast to musical wind instruments, in which the filter primarily determines f₀. Acoustic simulation further supported that gibbons' singing is produced analogously to professional human soprano singing, in which a precise tuning of the first formant (F₁) of the SVT to f₀ amplifies exclusively the f₀ component of the source. Thus, in gibbons, as in humans, dynamic control over the vocal tract configuration, rather than anatomical modifications, has been a dominant factor in determining call structure. The varied dynamic movements were adopted in response to unique social and ecological pressures in gibbons, allowing monogamous gibbons to produce pure‐tonal melodious songs in the dense tropical forests with poor visibility. Am J Phys Anthropol, 2012. © 2012 Wiley Periodicals, Inc.     

Ion‐dependent metabolic responses of Vicia faba L. to salt stress

Salt‐affected farmlands are increasingly burdened by chlorides, carbonates, and sulfates of sodium, calcium, and magnesium. Intriguingly, the underlying physiological processes are studied almost always under NaCl stress. Two faba bean cultivars were subjected to low‐ and high‐salt treatments of NaCl, Na₂SO₄, and KCl. Assimilation rate and leaf water vapor conductance were reduced to approximately 25–30% without biomass reduction after 7 days salt stress, but this did not cause severe carbon shortage. The equimolar treatments of Na⁺, K⁺, and Cl^? showed comparable accumulation patterns in leaves and roots, except for SO₄^2? which did not accumulate. To gain a detailed understanding of the effects caused by the tested ion combinations, we performed nontargeted gas chromatography–mass spectrometry‐based metabolite profiling. Metabolic responses to various salts were in part highly linearly correlated, but only a few metabolite responses were common to all salts and in both cultivars. At high salt concentrations, only myo‐inositol, allantoin, and glycerophosphoglycerol were highly significantly increased in roots under all tested conditions. We discovered several metabolic responses that were preferentially associated with the presence of Na⁺, K⁺, or Cl^?. For example, increases of leaf proline and decreases of leaf fumaric acid and malic acid were apparently associated with Cl^? accumulation.     

Escherichia coli populations adapt to complex,unpredictable fluctuations by minimizing trade‐offs across environments

In nature, organisms are simultaneously exposed to multiple stresses (i.e. complex environments) that often fluctuate unpredictably. Although both these factors have been studied in isolation, the interaction of the two remains poorly explored. To address this issue, we selected laboratory populations of Escherichia coli under complex (i.e. stressful combinations of pH, H₂O₂ and NaCl) unpredictably fluctuating environments for ~900 generations. We compared the growth rates and the corresponding trade‐off patterns of these populations to those that were selected under constant values of the component stresses (i.e. pH, H₂O₂ and NaCl) for the same duration. The fluctuation‐selected populations had greater mean growth rate and lower variation for growth rate over all the selection environments experienced. However, whereas the populations selected under constant stresses experienced trade‐offs in the environments other than those in which they were selected, the fluctuation‐selected populations could bypass the across‐environment trade‐offs almost entirely. Interestingly, trade‐offs were found between growth rates and carrying capacities. The results suggest that complexity and fluctuations can strongly affect the underlying trade‐off structure in evolving populations.     

Selection of symbiotically energy efficient strains of Rhizobium japonicum by their ability to induce a H2-uptake hydrogenase in the free-living state

The capacity of inducing a H₂-uptake hydrogenase in free-living cultures was examined in 21 strains of Rhizobium japonicum. Four strains were found to take up H₂ at rapid rates after 3 days of growth on agar slants inside sealed vials provided with an atmosphere of 5% H₂ in air. Soybean nodules from these strains lost little or no H₂ in air and their bacteroids oxidized H₂ at rates that were similar to those observed in free-living cultures. In contrast, three randomly chosen strains of R. japonicum that showed no H₂-uptake capacity in free-living state produced nodules which lost large amounts of H₂ and the corresponding bacteroids had no hydrogenase activity. A screening procedure is described for the selection of Rhizobium strains producing high energy-efficient nodules based on a test of their ability to induce a H₂-uptake hydrogenase in asymbiotic conditions.     

Differential Sensitivity of Macrocarpa and Microcarpa Types of Chickpea （Cicer arietinum L.） to Water Stress： Association of Contrasting Stress Response with Oxidative Injury

Chickpea (Cicer arietinum L.) is particularly sensitive to water stress at its reproductive phase and, under conditions of water stress, will abort flowers and pods, thus reducing yield potential. There are two types of chickpea: (i) Macrocarpa (“Kabuli”), which has large, rams head‐shaped, light brown seeds; and (ii) Microcarpa (“Desi”), which has small, angular and dark‐brown seeds. Relatively speaking, “Kabuli” has been reported to be more sensitive to water stress than “Desi”. The underlying mechanisms associated with contrasting sensitivity to water stress at the metabolic level are not well understood. We hypothesized that one of the reasons for contrasting water stress sensitivity in the two types of chickpea may be a variation in oxidative injury. In the present study, plants of both types were water stressed at the reproductive stage for 14 d. As a result of the stress, the “Kabuli” type exhibited an 80% reduction in seed yield over control compared with a 64% reduction observed for the “Desi” type. The decrease in leaf water potential (Ψ_w) was faster in the “Kabuli” compared with the “Desi” type. At the end of the water stress period, Ψ_w was reduced to ?2.9 and ?3.1 MPa in the “Desi” and “Kabuli” types, respectively, without any significant difference between them. On the last day of stress, “Kabuli” experienced 20% more membrane injury than “Desi”. The chlorophyll content and photosynthetic rate were significantly greater in “Desi” compared with “Kabuli”. The malondialdehyde and H₂O₂ content were markedly higher at the end of the water stress in “Kabuli” compared with “Desi”, indicating greater oxidative stress in the former. Levels of anti‐oxidants, such as ascorbic acid and glutathione, were significantly higher in “Desi” than “Kabuli”. Superoxide dismutase and catalase activity did not differ significantly between the two types of chickpea, whereas on the 10th day, the activities of ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase were higher in “Desi”. These findings indicate that the greater stress tolerance in the “Desi” type may be ascribed to its superior ability to maintain better water status, which results in less oxidative damage. In addition, laboratory studies conducted by subjecting both types of chickpea to similar levels of polyethylene glycol‐induced water stress and to 10 μ.mol/L abscisic acid indicated a greater capacity of the “Desi” type to deal with oxidative stress than the “Kabuli” type. (Managing editor: Ping He)     

Synthesis of 5-Alkyl-5-methyl-2-hydroxy-2-cyclopentenones

A hydrogen bacterium strain, N34, and its oxygen-resistant segregant strain, Y38, were subjected to a taxonomical study. Since both strains were capable of N₂-fixation, N₂-fixing facultative hydrogen autotrophs listed in “Bergey’s Manual of Systematic Bacteriology” were used for comparison. Both strains produced a water-insoluble carotenoid pigment, zeaxanthin dirhamnoside, indicating that both should be classified into the genus Xanthobacter. Then, the differential characteristics of the two species of the genus Xanthobacter, X. autotrophicus and X. flavus, were investigated as to both strains. The vitamin requirement, the sensitivity to oxygen under autotrophic conditions, the inducibility of hydrogenase, the substrate range of carbohydrates and N₂-fixing growth characteristics of both strains were almost completely opposite to those of X. flavus. Moreover, both strains coincided exactly with X. autotrophicus in morphological and other physiological characteristics. From these results both strains were identified as Xanthobacter autotrophicus.