bluuebunny/crossref_metadata_2025 huggingface dataset:
Phenotypic and genetic characterization of Paecilomyces lilacinus strains with biocontrol activity against root-knot nematodes
Authors: Gunasekera T S , Holland R J , Gillings M R , Briscoe D A , Neethling D C , Williams K L , Nevalainen K MH
Published: April 2011

Efficient selection of fungi for biological control of nematodes requires a series of screening assays. Assessment of genetic diversity in the candidate species maximizes the variety of the isolates tested and permits the assignment of a particular genotype with high nematophagous potential using a rapid novel assay. Molecular analyses also facilitate separation between isolates, allowing the identification of proprietary strains and trace biocontrol strains in the environment. The resistance of propagules to UV radiation is an important factor in the survival of a biocontrol agent. We have analyzed 15 strains of the nematophagous fungus Paecilomyces lilacinus using these principles. Arbitrarily primed DNA and allozyme assays were applied to place the isolates into genetic clusters, and demonstrated that some genetically related P. lilacinus strains exhibit widespread geographic distributions. When exposed to UV radiation, some weakly nematophagous strains were generally more susceptible than effective isolates. A microtitre tray-based assay used to screen the pathogenic activity of each isolate to Meloidogyne javanica egg masses revealed that the nematophagous ability varied between 37%-100%. However, there was no clear relationship between nematophagous ability and genetic clusters. Molecular characterizations revealed sufficient diversity to allow tracking of strains released into the environment.Key words: Paecilomyces lilacinus, LP-RAPD analysis, allozymes, UV sensitivity.
A CD-1 mouse model of infection with Staphylococcus aureus: Influence of gender on infection with MRSA and MSSA isolates
Authors: Yanke S J , Olson M E , Davies H D , Hart D A
Published: April 2011

Staphylococcus aureus is an important pathogen of humans and other animals, causing bacteremia, abscessation, toxemia, and other infectious diseases. An animal model using CD-1 mice was developed to study the pathogenesis of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA). When inoculated into the CD-1 mouse model, it was shown that both MSSA isolates, (HR 78 and CSA-1) and MRSA isolates (MRSA 456 and MRSA 457) led to chronic infection of the kidney. Female CD-1 mice inoculated with MRSA 456 proved to be more susceptible to infection and mortality than their male counterparts. Castrated mice became more susceptible to infection than intact male mice, suggesting a hormonal involvement in the infection process.Key words: Staphylococcus aureus, animal model, gender, MSSA, MRSA.
Isolation and characterization of highly thermophilic xylanolyticThermus thermophilusstrains from hot composts
Authors: Lyon Pierre-François , Beffa Trello , Blanc Michel , Auling Georg , Aragno Michel
Published: April 2011

This is the first detailed report of xylanolytic activity in Thermus strains. Two highly thermophilic xylanolytic bacteria, very closely related to non-xylanolytic T. thermophilus strains, have been isolated from the hottest zones of compost piles. Strain X6 was investigated in more detail. The growth rate (optical density monitoring) on xylan was 0.404·h-1at 75°C. Maximal growth temperature was 81°C. Xylanase activity was mainly cell-bound, but was solubilized into the medium by sonication. It was induced by xylan or xylose in the culture medium. The temperature and pH optima of the xylanases were determined to be around 100°C and pH 6, respectively. Xylanase activity was fairly thermostable; only 39% of activity was lost after an incubation period of 48 h at 90°C in the absence of substrate. Xylanolytic T. thermophilus strains could contribute to the degradation of hemicellulose during the thermogenic phase of industrial composting.Key words: Thermus, thermophilic aerobic bacteria, xylanase, thermostable enzyme, compost.
Identification of streptococci isolated from various sources by determination ofcfbgene and other CAMP-factor genes
Authors: Hassan Abdulwahed A , Abdulmawjood Amir , Yildirim Ali Ö , Fink Kristin , Lämmler Christoph , Schlenstedt Reglindis
Published: April 2011

In the present study, the CAMP-factor (cfb) gene of streptococci of serological group B (Streptococcus agalactiae) and the CAMP-factor (cfu) gene of S. uberis could be amplified by polymerase chain reaction. A cfb specific amplicon could be observed for all 128 phenotypically CAMP-positive S. agalactiae, for the phenotypically CAMP-negative S. agalactiae strain 74-360, and for 2 S. difficile reference strains. A cfu specific amplicon could be observed for all 7 phenotypically CAMP-positive S. uberis. Four S. agalactiae strains isolated from 4 cows with mastitis appeared to be phenotypically CAMP-negative and negative in the cfb gene PCR. The CAMP-positive and CAMP-negative isolates, including both S. difficile, could be identified as S. agalactiae by amplification of a S. agalactiae specific part of the V2 region of the 16S rRNA and a species-specific part of the 16S-23S rRNA intergenic spacer region. Amplification of an internal fragment of the cfb gene with a reduced annealing temperature yielded positive reactions not only for CAMP-positive S. agalactiae, but also for phenotypically CAMP-positive S. pyogenes (n = 4), S. canis (n = 28), and S. uberis (n = 7), indicating a close relation of the CAMP genes of these 4 species. The relation could be further demonstrated by sequencing the internal fragment of the CAMP-factor (cfg) gene of S. canis and comparing the sequence with those of S. agalactiae, S. pyogenes, and S. uberis.Key words: CAMP factor, cfb, cfu, S. canis.
Effects of mesophilic and thermophilic composts on suppression of Fusarium root and stem rot of greenhouse cucumber
Authors: Kannangara T , Utkhede R S , Paul J W , Punja Z K
Published: April 2011

Three composts were tested for their ability to suppress root and stem rot caused by the soil borne fungal pathogen Fusarium oxysporum f. sp. radicis-cucumerinum (FORC) on cucumber. Two of the composts were prepared from separated dairy solids either by windrow (WDS) or vermicomposting (VMC) while the third, obtained from International Bio-Recovery (IBR), was prepared from vegetable refuse using aerobic digestion. Three sets of potting mixes were prepared by mixing the composts with sawdust at varying ratios, and seeded with cucumber cv. Corona. After 14 days of growth in the greenhouse, inoculum of FORC (20 mL of 5 × 106micro-conidia per mL) was applied to each pot at three different times (14, 21, and 35 days). In unamended inoculated pots, the pathogen caused stunted growth and reduced flowers. Amendment of WDS in the potting mix suppressed these symptoms, while VMC and IBR had no effect. All three composts reduced the FORC colony forming units (cfu) at the end of the experiment (10 weeks). There was a large increase of fluorescent bacteria near the vicinity of roots particularly in WDS amended potting mixes. When water extracts of the composts were plated onto acidified potato dextrose agar (APDA), only IBR contained a potent thermostable inhibitor to FORC. This inhibitor was removed by activated charcoal but was not partitioned into petroleum ether at acid, basic, or neutral pH. Inhibition of FORC by IBR was not due to electrical conductivity or trace elements in the compost. Contrasting effectiveness of the WDS and VMC made from the same waste suggests that composting method can influence the disease suppression properties of the finished compost.Key words: Fusarium oxysporum f. sp. radicis-cucumerinum, biological control.
Transplantation and subsequent behavior of mitochondria in cells of Phytophthora
Authors: Gu Y H , Ko W H
Published: April 2011

Mitochondria isolated from streptomycin-resistant (Sr) protoplasts of Phytophthora parasitica were transferred into chloramphenicol-resistant (Cpr) protoplasts of P. parasitica or Phytophthora capsici with an average successful rate of 1.7 × 10-4, using a selective medium containing streptomycin. No colonies appeared when self-fusion products of donor mitochondria or recipient protoplasts were exposed to the selective medium. Mitochondria isolated from Cpr protoplasts of P. capsici were also transferred into Sr protoplasts of P. parasitica with a similar success rate using a selective medium containing chloramphenicol. Zoospores produced by the Cpr+Sr intraspecific mitochondrial hybrid gave rise to Sr and Cpr+Sr cultures. The second generation zoospores produced by Sr and Cpr+Sr cultures also gave rise to Sr and Cpr+Sr cultures, suggesting the possible occurrence of fusion between some of the Cpr mitochondria and Sr mitochondria, and the displacement of non-fused Cpr mitochondria in the receptor protoplast by the donor Sr mitochondria. Zoospores produced by the interspecific mitochondrial hybrid gave rise to Cpr, Sr, Cpr+Sr, and Cps +Ss cultures. The second generation zoospores produced by Cpr+Sr or Sr cultures also gave rise to the same four types of cultures, suggesting the existence of residual antibiotic-sensitive mitochondria (Cps+Ss) in the parental isolates and the random distribution of Cpr, Sr, and Cps+Ss mitochondria during asexual reproduction. Results suggest that the phenotype of antibiotic resistance / sensitivity was the end result of the interactions among the three types of mitochondria.Key words: mitochondrial transplantation, mitochondrial hybrid, antibiotic resistance, Phytophthora parasitica, Phytophthora capsici.
Genetic polymorphisms in three subtilisin-like protease isoforms (Pr1A, Pr1B, and Pr1C) from Metarhizium strains
Authors: Bidochka Michael J , Melzer Michael J
Published: April 2011

Restriction fragment length polymorphisms (RFLP) were examined in three isoforms of a gene family encoding subtilisin-like proteases (Pr1A, Pr1B, and Pr1C) in several isolates of the entomopathogenic fungus Metarhizium anisopliae. RFLP variation was not observed in any of the Pr1 genes from isolates within the same genetically related group. Between genetically related groups and between isolates from disparate geographical areas, the greatest variation in RFLP patterns was observed for Pr1A. When variation does occur at Pr1B and Pr1C, it was generally observed at an EcoRI site. Metarhizium anisopliae var. majus strain 473 and a M. flavoviride isolate were most dissimilar in RFLP patterns at all Pr1 genes when compared to the M. anisopliae strains. We suggest that Pr1 genes represent a gene family of subtilisin-like proteases and that the Pr1A gene encodes for the ancestral subtilisin-like protease which has subsequently duplicated and rearranged within the genome.Key words: Metarhizium anisopliae, protease, RFLP, entomopathogen.
Glutathione is a target in tellurite toxicity and is protected by tellurite resistance determinants inEscherichia coli
Authors: Turner Raymond J , Aharonowitz Yair , Weiner Joel H , Taylor Diane E
Published: April 2011

Tellurite (TeO32-) is highly toxic to most microorganisms. The mechanisms of toxicity or resistance are poorly understood. It has been shown that tellurite rapidly depletes the reduced thiol content within wild-type Escherichia coli. We have shown that the presence of plasmid-borne tellurite-resistance determinants protects against general thiol oxidation by tellurite. In the present study we observe that the tellurite-dependent depletion of cellular thiols in mutants of the glutathione and thioredoxin thiol:redox system was less than in wild-type cells. To identify the type of low-molecular-weight thiol compounds affected by tellurite exposure, the thiol-containing molecules were analyzed by reverse phase HPLC as their monobromobimane derivatives. Results indicated that reduced glutathione is a major initial target of tellurite reactivity within the cell. Other thiol species are also targeted by tellurite, including reduced coenzyme A. The presence of the tellurite resistance determinants kilA and ter protect against the loss of reduced glutathione by as much as 60% over a 2 h exposure. This protection of glutathione oxidation is likely key to the resistance mechanism of these determinants. Additionally, the thiol oxidation response curves were compared between selenite and tellurite. The loss of thiol compounds within the cell recovered from selenite but not to tellurite.Key words: tellurite, resistance, thiol oxidation, heavy metal toxicity, selenite, glutathione.
An endophyte of the tropical forage grass Brachiaria brizantha: Isolating, identifying, and characterizing the fungus, and determining its antimycotic properties
Authors: Kelemu Segenet , White Jr James F , Muñoz Fernando , Takayama Yuka
Published: April 2011

Brachiaria, predominantly an African genus, contains species, such as B. brizantha, an apomictic C4 grass, that are commercially important forage grasses in tropical America, where they now cover about 55 million hectares. From B. brizantha accession CIAT 6780, we isolated an endophytic fungus that may be economically significant. The fungus was identified as Acremonium implicatum (J. Gilman & E.V. Abott). 18S rDNA and ITS rDNA sequences were used to characterize isolates of the endophyte, and showed that they belonged to the Acremonium genus, being close to A. strictum and A. kiliense. Using the random amplified polymorphic DNA (RAPD) technique, involving arbitrary primers of 10 bases, we showed that the isolates were highly similar to each other. Antiserum produced from a monoconidial culture of A. implicatum isolated from B. brizantha 6780, differentiated the isolates consistently in line with the DNA data. When we compared endophyte-free with endophyte-infected B. brizantha CIAT 6780 plants, both artificially inoculated with the pathogenic Drechslera fungus, we found that the endophyte-infected plants had fewer and smaller lesions than did the endophyte-free plants. Sporulation of Drechslera sp. on artificially inoculated leaf sheath tissues was also much less on tissue infected with the endophyte.Key words: endophytes, Brachiaria, Acremonium, biological control.
Genetic diversity ofSinorhizobiumpopulations recovered from differentMedicagovarieties cultivated in Tunisian soils
Authors: Jebara M , Mhamdi R , Aouani M E , Ghrir R , Mars M
Published: April 2011

A collection of 468 rhizobial isolates was obtained from different ecological areas of Tunisia by trapping them on Medicago sativa cv. Gabes, Medicago scutelleta cv. Kelson, Medicago truncatula, and Medicago ciliaris. A subsample of 134 rhizobia was chosen to determine their plasmid profile, and 89 isolates were subjected to multilocus enzyme electrophoresis (MLEE) and PCR/RFLP analysis using 16S, IGS (inter genic spacer), and nifKD probes. Twenty-five representatives from these isolates were evaluated for their nodulation and nitrogen fixation capacities. MLEE studies revealed two groups with highly heterogeneous host specificity and geographical origin. The discriminatory power was found to be slightly better with the amplified ribosomal intergenic region, than the nifKD genes. Divisions detected by nifKD amplified DNA analysis matched those established by ribosomal PCR- RFLPs. The comparison between different analyses revealed that MLEE illustrated better phenotypic properties of isolates than PCR-RFLP or plasmid content analysis. Clear distinction between Sinorhizobium meliloti and Sinorhizobium medicae were observed by analysis of the IGS symbiotic regions between nifD and nifK genes. Were able to distinguish three inoculation groups; isolates trapped from M. sativa cv. Gabes and M. scutelleta cv. Kelson formed one inoculation group which was more closely related to isolates trapped from M. truncatula than those trapped from M. ciliaris.Key words: Sinorhizobium, Medicago, diversity, MLEE, PCR, symbiotic effectiveness.
Improving water stress tolerance of the biocontrol yeastCandida sakegrown in molasses-based media by physiological manipulation
Authors: Abadias M , Teixidó N , Usall J , Viñas I , Magan N
Published: April 2011

The biocontrol agent Candida sake was cultured on either an unmodified molasses-based medium (water activity, aw0.996) or on water stressed media produced by the addition of glycerol, glucose, NaCl, sorbitol, or proline to 0.98, and 0.96 awfor 24, 48, and 72 h, to study their impact on subsequent cell viability, and on concentrations of endogenous sugars (trehalose and glucose) and polyols (glycerol, erythritol, arabitol, and mannitol). The viability of cells of different ages cultured on these media was evaluated on NYDA medium with freely available water (aw0.995), and on medium modified with polyethylene glycol to aw0.95. Regardless of solute used, viable counts of cells grown on molasses-based medium (aw0.98) were equal to or higher than those obtained from the medium with water freely available. The amino acid proline stimulated growth at 10% concentration. In contrast, water stress induced by addition of NaCl, glucose, or sorbitol at aw0.96 caused a significant reduction in viable counts. Older cultures were more resistant to water stress. Glycerol and arabitol were the main solutes accumulated by C. sake cells in response to lowered aw. Intracellular concentration of these polyols depended more on the solute used to adjust the awthan on the awitself. Candida sake was more resistant to water stress with higher intracellular concentration of glycerol and erythritol.Key words: compatible solutes, polyols, sugars, improved viability, formulation.
Genetic diversity ofBradyrhizobiumstrains isolated fromArachis hypogaea
Authors: Saleena Lilly M , Loganathan P , Rangarajan Sunita , Nair Sudha
Published: April 2011

Rhizobia are used exclusively in agricultural systems for enhancing the ability of legumes to fix atmospheric nitrogen. Knowledge about the indigenous population is necessary for the selection and application of inoculant strains. In this study, we have assessed the genetic diversity of Bradyrhizobium strains isolated from the host plant, Arachis hypogaea along the coastline of Tamil Nadu. Different populations collected from varying environmental conditions were analysed for salt and pH tolerance. Genetic diversity among the strains was studied using RAPD markers and PCR-RFLP of 16S rDNA and nifD genes. The approaches used in this study yielded consistent results, which revealed a high degree of heterogeneity among strains and detection of two distinct genetic groups.Key words: rhizobia, diversity, salinity, RAPD, PCR-RFLP.
Effects of low temperature, cold shock, and various carbon sources on esterase and lipase activities and exopolysaccharide production by a psychrotrophicAcinetobactersp.
Authors: Barbaro S E , Trevors J T , Inniss W E
Published: April 2011

The activities of isocitrate lyase, esterase, and lipase by the psychrotrophic Acinetobacter sp. strain HH1-1 were monitored during incubation at 25°C, 5°C, and after a 25°C to 5°C down shift in growth temperature. During growth at 25°C, isocitrate lyase activity was detected in cell-free extracts, but at 5°C and after cold shock, activity was measured primarily in the cell culture supernatant. Strain HH1-1 produced two cell-associated esterases and an extracellular esterase and lipase. Activities of the extracellular esterase and lipase were reduced when cells were grown at 5°C and after cold shock. In contrast, an increased synthesis of a 53-kDa cell-associated esterase was observed 50 h after cold shock. An extracellular polysaccharide was also produced, indicated by a decrease in surface tension in cell culture supernatant when cells were incubated at 25°C; but like extracellular enzyme activity, production of the exopolymer was reduced when cells were subjected to low temperatures. These results indicated that the intracellular enzyme, isocitrate lyase, leaked out of the cell after cold shock and during growth at 5°C. The increased activity of a cell-associated esterase suggested this enzyme is required for growth at low temperatures. In contrast, activities of extracellular lipolytic enzymes and production of an extracellular polysaccharide were negatively affected at the lower temperatures.Key words: Acinetobacter, cold shock, esterase, lipase, exopolysaccharide.
IgA and IgM anti-Naegleria fowleriantibodies in human serum and saliva
Authors: Rivera Víctor , Hernández Dolores , Rojas Saúl , Oliver Gabriela , Serrano Jesús , Shibayama Mineko , Tsutsumi Víctor , Campos Rafael
Published: April 2011

Antibodies from IgA and IgM classes that recognize Naegleria fowleri (Nf) proteins were detected by the ELISA assay in serum and saliva from three groups of people: (i) subjects with upper respiratory tract infections (URTI) living in the parasite-endemic area, (ii) healthy persons from the same area, and (iii) healthy persons from a parasite-nonendemic area. In serum and in saliva the titers of IgA antibodies to Naegleria fowleri in the group of patients with URTI was significantly higher than that of the healthy group in the parasite-endemic area; also the titers of IgM antibodies in serum were significantly higher in patients. On the contrary, in saliva the antibodies were higher in healthy people from the parasite-endemic area. In all cases the subjects from the parasite-nonendemic area had lower antibody titers in serum and saliva.Key words: Naegleria fowleri, IgA, IgM antibodies, serum, saliva.
Mechanism of oxidation of inorganic sulfur compounds by thiosulfate-grown Thiobacillus thiooxidans
Authors: Masau Rosemarie Jefferey Y , Oh Jae Key , Suzuki Isamu
Published: April 2011

Thiobacillus thiooxidans was grown at pH 5 on thiosulfate as an energy source, and the mechanism of oxidation of inorganic sulfur compounds was studied by the effect of inhibitors, stoichiometries of oxygen consumption and sulfur, sulfite, or tetrathionate accumulation, and cytochrome reduction by substrates. Both intact cells and cell-free extracts were used in the study. The results are consistent with the pathway with sulfur and sulfite as the key intermediates. Thiosulfate was oxidized after cleavage to sulfur and sulfite as intermediates at pH 5, the optimal growth pH on thiosulfate, but after initial condensation to tetrathionate at pH 2.3 where the organism failed to grow. N-Ethylmaleimide (NEM) inhibited sulfur oxidation directly and the oxidation of thiosulfate or tetrathionate indirectly. It did not inhibit the sulfite oxidation by cells, but inhibited any reduction of cell cytochromes by sulfur, thiosulfate, tetrathionate, and sulfite. NEM probably binds sulfhydryl groups, which are possibly essential in supplying electrons to initiate sulfur oxidation. 2-Heptyl-4-hydroxy-quinoline N-oxide (HQNO) inhibited the oxidation of sulfite directly and that of sulfur, thiosulfate, and tetrathionate indirectly. Uncouplers, carbonyl cyanide-m-chlorophenylhydrazone (CCCP) and 2,4-dinitrophenol (DNP), inhibited sulfite oxidation by cells, but not the oxidation by extracts, while HQNO inhibited both. It is proposed that HQNO inhibits the oxidation of sulfite at the cytochrome b site both in cells and extracts, but uncouplers inhibit the oxidation in cells only by collapsing the energized state of cells, ΔµH+, required either for electron transfer from cytochrome c to b or for sulfite binding.Key words: Thiobacillus thiooxidans, thiosulfate, oxidation, sulfite.
Quantification of autolysis inPenicillium chrysogenumby semiautomated image analysis
Authors: McIntyre M , Eade J K , Cox P W , Thomas C R , White S , Berry D R , McNeil B
Published: April 2011

An image analysis method is described for the characterization of empty (autolyzed and inactive) regions within the mycelia of filamentous fungi. It extends a previous method that characterized only regions filled with cytoplasm or vacuoles (i.e., the active biomass). The method is semiautomatic, requiring some manual editing before automated measurements. When the method was used for samples from a batch fermentation of an industrial strain of Penicillium chrysogenum, the empty regions were observed to constitute up to 15% (by projected area) of the biomass during the growth phase. After nutrient exhaustion, however, the proportion of empty regions rose rapidly, eventually representing more than 50% of the biomass by the end of fermentation. The increase in the percentage of empty regions coincided with a decrease in biomass (as measured by dry cell weight) and a fall in penicillin titre. Further morphological analysis revealed that fragmentation of mycelia, particularly clumps, coincided with increases in the levels of empty regions. This new image analysis method gave additional information on hyphal differentiation and a measure of autolysis. It was also a useful indicator of the processes leading to autolysis.Key words: autolysis, penicillin, differentiation, double grey.
Uridylylation of the PII protein fromHerbaspirillum seropedicae
Authors: Benelli Elaine M , Buck Martin , Souza Emanuel Maltempi de , Yates Marshall Geoffrey , Pedrosa Fabio O
Published: April 2011

The PII protein is apparently involved in the control of NifA activity in Herbaspirillum seropedicae. To evaluate the probable role of PII in signal transduction, uridylylation assays were conducted with purified H. seropedicae PII and Escherichia coli GlnD, or a cell-free extract of H. seropedicae as sources of uridylylating activity. The results showed that α-ketoglutarate and ATP stimulate uridylylation whereas glutamine inhibits uridylylation. Deuridylylation of PII-UMP was dependent on glutamine and inhibited by ATP and α-ketoglutarate. PII uridylylation and (or) deuridylylation in response to these effectors suggests that PII is a nitrogen level signal transducer in H. seropedicae.Key words: nitrogen regulation, uridylylation, PII protein, Herbaspirillum seropedicae.
Study of lipase-catalyzed hydrolysis of some monoterpene esters
Authors: Chatterjee T , Chatterjee B K , Bhattacharyya D K
Published: April 2011

Studies of the hydrolysis of bornyl, citronellyl, geranyl, and terpenyl acetates by commercially available lipases of Candida rugosa, Rhizopus arrhizus, and Mucor miehei are presented. The hydrolysis of these monoterpene esters is investigated at various temperatures and pHs, and the time dependence of the percentage of esters hydrolysed is studied. The catalytic activities of the lipases in hydrolysing the esters are compared and, overall, it is observed that under the experimental conditions used the nonspecific lipase from C. rugosa produces the highest yields of the monoterpene alcohols in comparison to the primary-ester specific lipases such as R. arrhizus and M. miehei. A rate kinetic model has been used to understand the time dependence of the yield of the product acid.Key words: enzymatic hydrolysis, kinetics, lipase, monoterpene esters.
Acid and alkaline treatments for enhancing the growth of rhizobia in sludge
Authors: Rebah Faouzi Ben , Tyagi Rajeshwar D , Prévost Danielle
Published: April 2011

Wastewater sludges have been proposed as an effective media for the production of rhizobia. The effect of total suspended solid (TSS) concentrations and pretreatments of sludge on the growth of Sinorhizobium meliloti were investigated. Acid (pH 2.06.0 obtained with H2SO4) and alkaline (50200 mequiv.wt./L of NaOH) treatments were applied to enhance the biodegradability of primary (0.325%3.2% TSS obtained by dilution of original sample) and secondary (0.2%0.4% TSS obtained by concentration of original sample) sludges. In primary sludge without pretreatment, the highest cell count (11.10 × 109 cfu/mL) was obtained with 1.3% TSS. However, a maximum cell count of 13.00 × 109 cfu/mL was reached using an acid treatment of pH 2.0 and a 0.325% TSS concentration. Moreover, the alkaline treatment with 100 mequiv.wt./L of NaOH and 0.65% TSS increased the cell yield to 21.00 × 109 cfu/mL. For secondary sludge without pretreatment, no enhancement of growth was observed while increasing TSS concentration. This may be due to the increase of inhibitory substances, such as heavy metals, and of the Ca and Mg concentrations. As in primary sludge, some acid and alkaline treatments of secondary sludge tend to improve the cell count of S. meliloti. However, the highest value of 9.80 × 109 cfu/mL obtained with 0.4% TSS at pH 2.0 was lower than that obtained with primary sludge. It was also observed that S. meliloti grown in treated sludges maintained its capacity to nodulate alfalfa.Key words: wastewater sludge, rhizobium, inoculum, sludge pretreatment, nodulation.
Diversity in the rhizobia associated withPhaseolus vulgarisL. in Ecuador, and comparisons with Mexican bean rhizobia
Authors: Bernal Gustavo , Graham Peter H
Published: April 2011

Common beans (Phaseolus vulgaris L.) have centers of origin in both Mesoamerica and Andean South America, and have been domesticated in each region for perhaps 5000 years. A third major gene pool may exist in Ecuador and Northern Peru. The diversity of the rhizobia associated with beans has also been studied, but to date with an emphasis on the Mesoamerican center of origin. In this study we compared bean rhizobia from Mexico and Andean South America using both phenotypic and phylogenetic approaches. When differences between the rhizobia of these two regions were shown, we then examined the influence of bean cultivar on the most probable number (MPN) count and biodiversity of rhizobia recovered from different soils. Three clusters of bean rhizobia were distinguished using phenotypic analysis and principal-component analysis of Box A1R-PCR banding patterns. They corresponded principally to isolates from Mexico, and the northern and southern Andean regions, with isolates from southern Ecuador exhibiting significant genetic diversity. Rhizobia from Dalea spp., which are infective and effective on beans, may have contributed to the apparent diversity of rhizobia recovered from the Mesoamerican region, while the rhizobia of wild Phaseolus aborigineus from Argentina showed only limited similarity to the other bean rhizobia tested. Use of P. vulgaris cultivars from the Mesoamerican and Andean Phaseolus gene pools as trap hosts did not significantly affect MPN counts of bean rhizobia from the soils of each region, but did influence the diversity of the rhizobia recovered. Such differences in compatibility of host and Rhizobium could be a factor in the poor reputation for nodulation and N2fixation in this crop.Key words: Phaseolus vulgaris, Rhizobium diversity, Dalea spp., soil populations, specificity, centers of origin.
Rhizobial survival and nodulation of chickpea as influenced by fungicide seed treatment
Authors: Kyei-Boahen S , Slinkard A E , Walley F L
Published: April 2011

The survival of Rhizobium ciceri on chickpea (Cicer arietinum cv. Myles) seed, treated separately with 1 of 4 commercial fungicides, i.e., Apron®, Arrest 75W®, Crown®, or Captan®, was examined under laboratory conditions using standard serial dilution and plate count techniques. The resulting effects of fungicideRhizobium interactions on nodulation, N2 fixation, and plant growth were assessed in a controlled environment. Fungicide treatment decreased the number of viable rhizobia on the seed. In general, the toxicity of the fungicides in terms of rhizobial viability increased in the following order: Control = Crown < Arrest = Apron < Captan. Although Crown had no effect on rhizobial viability assessed under laboratory conditions, it significantly reduced nodulation, percent N derived from the atmosphere (%Ndfa), and shoot dry matter. Seed treated with Arrest and Captan decreased nodule dry weight and %Ndfa, but only Arrest reduced dry matter yield. Apron had no effect on any of the parameters measured at the early pod-filling stage and was compatible with the chickpea inoculum used in this study.Key words: fungicides, Rhizobium ciceri, inoculation, nodulation, chickpea, Cicer arietinum.
Biotransformation and partial mineralization of the explosive 2,4,6-trinitrotoluene (TNT) by rhizobia
Authors: Labidi Mohamed , Ahmad Darakhshan , Halasz Annamaria , Hawari Jalal
Published: April 2011

Three strains, T10, B5, and M8, each belonging to a different species of the family Rhizobiaceae and isolated from atrazine-contaminated soils, were tested for their ability to transform 2,4,6-trinitrotoluene (TNT) (50 µg·mL1) in liquid cultures using glucose as the C-source. All three strains were able to transform TNT to hydroxy lamino dinitro toluenes (2-HADNT, 4-HADNT), aminodinitrotoluenes (2-ADNT, 4-ADNT), and diaminonitrotoluene (2,4-DANT). The transformation was significantly faster in the presence of glutamate. Furthermore, the major metabolites that accumulated in cultures were 2-ADNT with glucose, and 4-ADNT with glutamate plus glucose. Rhizobium trifolii T10 was also tested for its ability to transform high levels of TNT (~350 µg·mL1) in a soil slurry. Almost 60% of the TNT was transformed within 2 days in bioaugmented soil slurries, and up to 90% when cultures were supplemented with glucose and glutamate. However, mineralization was minimal in all cases, less than 2% in 78 days. This is the first report on the degradation of TNT by rhizobial strains, and our findings suggest that rhizobia have the potential to play an important role in the safe decontamination of soils and sites contaminated with TNT if bioaugmentation with rhizobia is shown to have no ecotoxicological consequence.Key words: Rhizobium, 2,4,6-trinitrotoluene (TNT), biotransformation.
Use of β-D-galactosidase and β-D-glucuronidase activities for quantitative detection of total and fecal coliforms in wastewater
Authors: George Isabelle , Crop Philippe , Servais Pierre
Published: April 2011

Two enzymatic methods based on the measurement of the β-D-galactosidase activity of total coliforms and the β-D-glucuronidase activity of Escherichia coli were used to assess coliform levels in wastewater alongside traditional culture-based techniques, which can be biased by the aggregation of bacteria or their attachment to particles. Enzymatic methods were precise (i.e., coefficients of variation were 9%15%), rapid (response in 20 min), and correlated well (in log units) with traditional techniques for raw and treated sewage (r2> 0.75). They were used for rapid assessment of coliform removal efficiency in two different wastewater treatments. These methods could be useful for the estimation of the abundance of coliforms in domestic sewage and their removal by wastewater treatment processes.Key Words: wastewater, total coliforms, fecal coliforms, β-D-galactosidase activity, β-D-glucuronidase activity.
Effets de l'inoculation avec des souches deRhizobium leguminosarumbiovartrifoliisur la croissance du blé dans deux sols du Maroc
Authors: Hilali Abdelaly , Prévost Danielle , Broughton William J , Antoun Hani
Published: April 2011

One hundred strains of Rhizobium leguminosarum bv. trifolii were isolated from roots of wheat cultivated in rotation with clover in two different regions of Morocco. The isolates were first screened for their effect on the growth of the cultivar Rihane of wheat cultivated in an agricultural soil under greenhouse conditions. After 5 weeks of growth, 14 strains stimulating the fresh or dry matter yield of shoots were selected and used in a second pot inoculation trial performed with two different agricultural soils. The results show that the strains behaved differently according to the soil used. In the loamy sand Rabat, strain IAT 168 behaved potentially like a plant growth promoting rhizobacteria (PGPR), as indicated by the 24% increases (P < 0.1) observed in wheat shoot dry matter and grain yields. In the silty clay Merchouch, no PGPR activity was observed, and 6 strains showed a significant deleterious effect on yields. These observations suggest that it is very important in a crop rotation system to choose a R. leguminosarum bv. trifolii strain that is effective with clover and shows PGPR activity with wheat to avoid deleterious effects on wheat yields.Key words: deleterious bacteria, PGPR (plant growth promoting rhizobacteria), Trifolium alexandrinum, Triticum aestivum.
Precise excision of bacteriophage Mu DNA
Authors: Abbes Chiraz , Sezonov Guennadi , Joseleau-Petit Danièle , D'Ari Richard , Liébart Jean-Claude
Published: April 2011

The temperate bacteriophage Mu is a transposable element that can integrate randomly into bacterial DNA, thereby creating mutations. Mutants due to an integrated Mu prophage do not give rise to revertants, as if Mu, unlike other transposable elements, were unable to excise precisely. In the present work, starting with a lacZ::Muc62(Ts) strain unable to form Lac+colonies, we cloned a lacZ+gene in vivo on a mini-Mu plasmid, under conditions of prophage induction. In all lac+plasmids recovered, the wild-type sequence was restored in the region where the Mu prophage had been integrated. The recovery of lacZ+genes shows that precise excision of Mu does indeed take place; the absence of Lac+colonies suggests that precise excision events are systematically associated with loss of colony-forming ability.Key words: transposable element, Escherichia coli, mini-Muduction.
Creation of hybrid vigor through nuclear transplantation in Phytophthora
Authors: Gu Yu-Huan , Ko Wen-Hsiung
Published: April 2011

When isolated nuclei of a diploid oomycete, Phytophthora parasitica, were fused with protoplasts of another strain of the same species, the regenerated nuclear hybrids grew faster than the parental isolates. Such a phenomenon did not occur in hybrids regenerated from mitochondrionprotoplast or protoplastprotoplast fusion products between these two strains. These results indicate that hybrid vigor is the result of the interaction between two different kinds of nuclei, but not between mitochondria, and they suggest that the presence of mitochondria from nuclear donor cells represses the expression of increased vigor. The nuclear hybrids also expressed increased fungicide resistance and propagule production. Increased vigor in growth was also observed in the interspecific nuclear hybrids when isolated nuclei of P. parasitica were transferred into protoplasts of Phytophthora capsici, and vice versa. This phenomenon may have potential applications, such as the creation of superior fungal strains and plant cultivars with improved commercial traits for usage in industry and agriculture.Key words: hybrid vigor, nuclear transplantation, Phytophthora parasitica, Phytophthora capsici.
Rapid identification of Escherichia coli microcin J25 producing strains using polymerase chain reaction and colony blot hybridization
Authors: Duarte Mariela , Cottenceau Gilles , Portrait Véronique , Pons Anne-Marie
Published: April 2011

To screen, isolate, and characterize bacterial populations producing microcin J25, we report here two rapid, reliable, and sensitive methods, using polymerase chain reaction and colony blot hybridization with a digoxigenin-labelled probe. A sample of 26 Escherichia coli strains isolated from poultry intestinal contents was evaluated to detect the sequence of mcjA, the gene encoding the MccJ25 precursor. The two molecular techniques were compared with the commonly used cross-immunity tests. They generate accurate data with no obvious cross-reactions with other microcins. The results display that the producers of MccJ25 were widely distributed in the poultry intestinal habitat. The applications of these molecular methods will be useful in future studies of microcinogenic populations, and thus contribute to understand the relationships within the complex intestinal microbial ecosystem.Key words: microcin J25, microcinogenic strains detection, digoxigenin-labelled probe, colony hybridization, polymerase chain reaction.
Comparison of a nested polymerase chain reaction  restriction fragment length polymorphism method, the PATH antigen detection method, and microscopy for the detection and identification of malaria parasites
Authors: Parkes Richard , Lo Teresa , Wong Quantine , Isaac-Renton Judith L , Byrne Sean K
Published: April 2011

A nested polymerase chain reaction  restriction fragment length polymorphism (PCRRFLP) method, the PATH antigen detection method, and light microscopy were compared for their capacity to detect and identify Plasmodium species. One hundred and thirty-six blood specimens obtained from patients suspected of having malaria were examined by each of the three methods. Forty-four specimens were positive for malaria using microscopy as the "gold standard". The sensitivity for nested PCR was 100%, and the specificity was 98%. For the detection of Plasmodium falciparum, the antigen detection method had a sensitivity of 100% and a specificity of 97%. Species identification obtained using PCRRFLP was identical or superior to light microscopy in 42 cases (96%). Although the nested PCRRFLP method was more sensitive and specific, the rapid turnaround time and high sensitivity of the antigen detection method makes it a useful adjunct to standard microscopy.Key words: malaria, PCRRFLP, antigen detection.
Phenotypic and genotypic characterization of esterase-producingUreibacillus thermosphaericusisolated from an aerobic digestor of swine waste
Authors: Gagné A , Chicoine M , Morin A , Houde A
Published: April 2011

Eight closely related thermophilic strains were isolated from an aerobic and thermophilic treatment of swine wastes. The pleomorphic cells (short and long rods; cocci) showed peritrichous flagella, terminally swollen sporangium, and liberated spores exhibiting hairy appendages. The Gram reaction was negative for both young (4 h) and old (48 h) cultures. Several features, such as colonial morphology, growth between 35°C and 65°C, presence of catalase, presence of spores, and strictly aerobic metabolism (except for one strain), are similar to those found for the genus Bacillus. The inability of the strains to use sugars, except esculin, as source of carbon and energy and the whole cell fatty acid composition are similar to those found in Bacillus thermosphaericus DSM 10633. Sequence analysis of the 16S rRNA gene revealed 99.8%99.9% identity for seven of the thermophilic strains with this species. A new genus, Ureibacillus, was recently proposed for type strain B. thermosphaericus DSM 10633 The last strain exhibits 97.8% and 97.3% identity with Ureibacillus terrenus DSM12654 and Bacillus sp. TP-84, respectively. Esterase activities were detected for all strains, and assays on p-nitrophenyl butyrate and p-nitrophenyl caprylate revealed that strains were more active on the shorter substrate.Key words: Ureibacillus thermosphaericus, thermophilic, esterase.
Phosphite disrupts the acclimation of Saccharomyces cerevisiae to phosphate starvation
Authors: McDonald Allison E , Niere Julie O , Plaxton William C
Published: April 2011

The influence of phosphite (H2PO3) on the response of Saccharomyces cerevisiae to orthophosphate (HPO42; Pi) starvation was assessed. Phosphate-repressible acid phosphatase (rAPase) derepression and cell development were abolished when phosphate-sufficient (+Pi) yeast were subcultured into phosphate-deficient (Pi) media containing 0.1 mM phosphite. By contrast, treatment with 0.1 mM phosphite exerted no influence on rAPase activity or growth of +Pi cells. 31P NMR spectroscopy revealed that phosphite is assimilated and concentrated by yeast cultured with 0.1 mM phosphite, and that the levels of sugar phosphates, pyrophosphate, and particularly polyphosphate were significantly reduced in the phosphite-treated Pi cells. Examination of phosphite's effects on two PHO regulon mutants that constitutively express rAPase indicated that (i) a potential target for phosphite's action in Pi yeast is Pho84 (plasmalemma high-affinity Pi transporter and component of a putative phosphate sensor-complex), and that (ii) an additional mechanism exists to control rAPase expression that is independent of Pho85 (cyclin-dependent protein kinase). Marked accumulation of polyphosphate in the Δpho85 mutant suggested that Pho85 contributes to the control of polyphosphate metabolism. Results are consistent with the hypothesis that phosphite obstructs the signaling pathway by which S. cerevisiae perceives and responds to phosphate deprivation at the molecular level.Key words: Saccharomyces cerevisiae, phosphite, phosphate starvation, PHO regulon.
Binding ofEscherichia coliverotoxins to cell surface protein on wild-type and globotriaosylceramide-deficient Vero cells
Authors: Devenish John , Gyles Carlton , LaMarre Jonathan
Published: April 2011

We have examined verotoxin (VT) binding to cell surface proteins. When Vero or globotriaosylceramide (Gb3) deficient Vero (VRP) cells were incubated with125I-labelled verotoxin 2 (VT2) and disuccinimidyl suberate cross-linker, SDS-PAGE of cell lysates showed radiolabelled bands at 44, 50, 60, 86, 102, and 138 kDa. When125I-labelled verotoxin 1 (VT1) was cross-linked, radioactive bands occurred at 51, 67, 101, 160, 188, and 232 kDa. In contrast,125I-labelled VT1 B subunit produced a single radioactive band migrating at 50 kDa. CHO cells did not bind labelled VT. VT2 binding to VRP cells fit a rectangular hyperbola suggesting a single class of binding sites. In contrast, VT1 and VT1 B subunit binding to VRP cells was best fit by sigmoidal curves suggesting the presence of positive cooperativity between at least two binding sites. Scatchard analysis of VT2 binding data yielded 3.5 times 109molecules bound/ µg of cell protein with an equilibrium dissociation constant (KD) of 13 nM. The apparent KDwas 9.7 nM for VT1 and 73.2 nM for VT1 B subunit. These results indicate that VT binds to a protein, or proteins, on the surface of susceptible cells and that there appear to be differences between VT1 and VT2 binding. Interactions between VT1 or VT2 and the proteins demonstrated here may be important in the biological activity of VT.Key words: verotoxin, protein receptors, hemolytic uremic syndrome, Escherichia coli.
Antifungal activity of chitinolytic bacteria isolated from airtight stored cereal grain
Authors: Frändberg Emma , Schnürer Johan
Published: April 2011

Chitinolytic bacteria are used as biocontrol agents of plant pathogenic fungi. They might also potentially inhibit growth of molds, e.g., Aspergillus spp. and Penicillium spp., in stored plant material. We isolated chitinolytic bacteria from airtight stored cereal grain and evaluated their antifungal capacity. Between 0.01 and 0.5% of the total aerobic counts were chitinolytic bacteria. Gram-negative bacteria, mainly Pseudomonadaceae, constituted approximately 80% of the chitinolytic population. Gram-positive isolates belonged predominantly to the Corynebacterium-Arthrobacter group, Streptomyces, and Bacillus. Chitinolytic activity was evaluated using culture filtrates from chitin-grown isolates as the release of p-nitrophenol from p-nitrophenyl N,N'-diacetylchitobiose and as the formation of clearing zones on chitin agar. No correlation between chitinolytic activity and antifungal effects was found when challenging Penicillium roqueforti Dierckx with bacterial isolates on chitin agar in a dual culture bioassay. Fungal hyphae frequently grew seemingly unaffected through the bacterial colony of a high chitinase producer on colloidal chitin. Only 4% of the chitinolytic isolates had strong effects on fungal growth. Among these, Streptomyces halstedii (K122) and Streptomyces coelicolor (K139) inhibited growth of a broad range of fungi. Streptomyces halstedii affected hyphal morphology and decreased the radial growth rate of all fungi investigated. These effects were not caused by volatile metabolites, polyenes, or N-carbamoyl-D-glucosamine.Key words: antifungal, chitinase, Streptomyces halstedii, Streptomyces coelicolor.