bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of sulfate concentration in dechlorination of 3,4,5-trichlorocatechol by stable enrichment cultures grown with coumarin and flavanone glycones and aglycones
Authors: Allard A S , Hynning P A , Remberger M , Neilson A H
Published: January 2020

Metabolically stable anaerobic enrichment cultures have been obtained from sediment samples contaminated with chlorophenolic compounds. Enrichment was carried out with esculin, esculetin, naringin, naringenin, fraxin, quercetin, and acetate in media with two sulfate concentrations. These cultures were used to examine the O-demethylation of 4,5,6-trichloroguaiacol and the dechlorination of 3,4,5-trichlorocatechol. Whereas O-demethylation was observed in all cultures, the occurrence of dechlorination was significantly more restricted. The presence of the carbohydrate moiety in the cultures enriched with the glycones repressed development of populations which were able to carry out dechlorination. Although sulfate at a concentration of 2 g/liter in the primary enrichments blocked the development of populations able to bring about dechlorination, addition of sulfate at this concentration did not inhibit dechlorination in cultures possessing this capability. Different dichlorocatechol isomers were produced under the various conditions, so that in view of the established resistance of some of these to further dechlorination, the ultimate fate of 3,4,5-trichlorocatechol in the natural environment remains partly unresolved. No enrichment culture containing a low sulfate concentration was able to dechlorinate either 2,4,5-trichlorophenol or 2,4,6-trichlorobenzoate.
Lignocellulose Degradation during Solid-State Fermentation:
Pleurotus ostreatus
versus
Phanerochaete chrysosporium
Authors: Kerem Zohar , Friesem Dana , Hadar Yitzhak
Published: January 2020

Lignocellulose degradation and activities related to lignin degradation were studied in the solid-state fermentation of cotton stalks by comparing two white rot fungi,
Pleurotus ostreatus
and
Phanerochaete chrysosporium. P. chrysosporium
grew vigorously, resulting in rapid, nonselective degradation of 55% of the organic components of the cotton stalks within 15 days. In contrast,
P. ostreatus
grew more slowly with obvious selectivity for lignin degradation and resulting in the degradation of only 20% of the organic matter after 30 days of incubation. The kinetics of
14
C-lignin mineralization exhibited similar differences. In cultures of
P. chrysosporium
, mineralization ceased after 18 days, resulting in the release of 12% of the total radioactivity as
14
CO
2
. In
P. ostreatus
, on the other hand, 17% of the total radioactivity was released in a steady rate throughout a period of 60 days of incubation. Laccase activity was only detected in water extracts of the
P. ostreatus
fermentation. No lignin peroxidase activity was detected in either the water extract or liquid cultures of this fungus. 2-Keto-4-thiomethyl butyric acid cleavage to ethylene correlated to lignin degradation in both fungi. A study of fungal activity under solid-state conditions, in contrast to those done under defined liquid culture, may help to better understand the mechanisms involved in lignocellulose degradation.
Analysis of proteins associated with growth of Bacteroides ovatus on the branched galactomannan guar gum
Authors: Valentine P J , Salyers A A
Published: January 2020

Bacteroides ovatus, a gram-negative obligate anaerobe from the human colon, can ferment the branched galactomannan guar gum. Previously, three enzymes involved in guar gum breakdown were characterized. The expression of these enzymes appeared to be regulated; i.e., specific activities were higher in extracts from bacteria grown on guar gum than in extracts from bacteria grown on the monosaccharide constituents of guar gum, mannose and galactose. In the present study, we used two-dimensional gel analysis to determine the total number of B. ovatus proteins enhanced during growth on guar gum. Twelve soluble proteins and 20 membrane proteins were expressed at higher levels in guar gum-grown cells than in galactose-grown cells. An unexpected finding was that the expression of the two galactomannanases was induced by glucose as well as guar gum. Three other proteins, one membrane protein and two soluble proteins, had this same expression pattern. The remainder of the guar gum-associated proteins seen on two-dimensional gels and the guar gum-associated alpha-galactosidase were induced in cells grown on guar gum but not in cells grown on glucose. Two transposon-generated mutants (M-5 and M-7) that could not grow on guar gum were isolated. Both mutants still expressed the galactomannanases and the alpha-galactosidase. They also still expressed all of the guar gum-associated proteins that could be detected in two-dimensional gels of glucose-grown or galactose-grown cells. A second transposon insertion that suppressed the guar gum-negative phenotype of M-5 was isolated and characterized. The characteristics of this suppressor mutant indicated that the original transposon insertion was probably in a regulatory locus.
Dipstick immunoassay to detect enterohemorrhagic Escherichia coli O157:H7 in retail ground beef
Authors: Kim M S , Doyle M P
Published: January 2020

A sensitive and easy-to-perform dipstick immunoassay to detect Escherichia coli O157:H7 in retail ground beef was developed by using a sandwich-type assay (with a polyclonal antibody to E. coli O157 as the capture antibody and a monoclonal antibody to E. coli O157:H7 as the detection antibody) on a hydrophobic polyvinylidine difluoride-based membrane. E. coli O157:H7 in ground beef could be detected within 16 h, including incubation for 12 h in enrichment broth and the immunoassay, which takes 4 h. Pure culture cell suspensions of 10(5) or 10(6) E. coli O157:H7 organisms per ml produced intense color reactions in the immunoassay, whereas faint but detectable reactions occurred with 10(3) CFU/ml. The sensitivity of the combined enrichment-immunoassay procedure as determined by using ground beef inoculated with E. coli O157:H7 was 0.1 to 1.3 cells per g, with a false-positive rate of 2.0%. A survey of retail ground beef using this procedure revealed that 1 of 76 samples was contaminated by E. coli O157:H7.
Molybdenum Oxidation by
Thiobacillus ferrooxidans
Authors: Sugio Tsuyoshi , Hirayama Kouichi , Inagaki Kenji , Tanaka Hidehiko , Tano Tatsuo
Published: January 2020

Thiobacillus ferrooxidans
AP19-3 oxidized molybdenum blue (Mo
5+
) enzymatically. Molybdenum oxidase in the plasma membrane of this bacterium was purified ca. 77-fold compared with molybdenum oxidase in cell extract. A purified molybdenum oxidase showed characteristic absorption maxima due to reduced-type cytochrome oxidase at 438 and 595 nm but did not show absorption peaks specific for
c
-type cytochrome. The optimum pH of molybdenum oxidase was 5.5. The activity of molybdenum oxidase was completely inhibited by sodium cyanide (5 mM) or carbon monoxide, and an oxidized type of cytochrome oxidase in a purified molybdenum oxidase was reduced by molybdenum blue, indicating that cytochrome oxidase in the enzyme plays a crucial role in molybdenum blue oxidation.
High-pressure, high-temperature bioreactor for comparing effects of hyperbaric and hydrostatic pressure on bacterial growth
Authors: Nelson C M , Schuppenhauer M R , Clark D S
Published: January 2020

We describe a high-pressure reactor system suitable for simultaneous hyperbaric and hydrostatic pressurization of bacterial cultures at elevated temperatures. For the deep-sea thermophile ES4, the growth rate at 500 atm (1 atm = 101.29 kPa) and 95 degrees C under hydrostatic pressure was ca. three times the growth rate under hyperbaric pressure and ca. 40% higher than the growth rate at 35 atm.
Molecular analyses of the lactococcin A gene cluster from Lactococcus lactis subsp. lactis biovar diacetylactis WM4
Authors: Stoddard G W , Petzel J P , van Belkum M J , Kok J , McKay L L
Published: January 2020

The genes responsible for bacteriocin production and immunity in Lactococcus lactis subsp. lactis biovar diacetylactis WM4 were localized and characterized by DNA restriction fragment deletion, subcloning, and nucleotide sequence analysis. The nucleotide sequence of a 5.6-kb AvaII restriction fragment revealed a cluster with five complete open reading frames (ORFs) in the same orientation. DNA and protein homology analyses, combined with deletion and Tn5 insertion mutagenesis, implicated four of the ORFs in the production of and immunity to lactococcin A. The last two ORFs in the cluster were the lactococcin A structural and immunity genes, lcnA and lciA. The two ORFs immediately upstream of lcnA and lciA were designated lcnC and lcnD, and the proteins that they encoded showed similarities to proteins of signal sequence-independent secretion systems. lcnC encodes a protein of 716 amino acids that could belong to the HlyB family of ATP-dependent membrane translocators. LcnC contains an ATP binding domain in a conserved C-terminal stretch of approximately 200 amino acids and three putative hydrophobic segments in the N terminus. The lcnD product, LcnD, of 474 amino acids, is essential for lactococcin A expression and shows structural similarities to HlyD and its homologs. On the basis of these results, a secretion apparatus that is essential for the full expression of active lactococcin A is postulated.
Evaluation of the Organon-Teknika MICRO-ID LISTERIA system
Authors: Bannerman E , Yersin M N , Bille J
Published: January 2020

The MICRO-ID LISTERIA system, designed to identify Listeria isolates to species level within 24 h, was compared with conventional biochemical identification. MICRO-ID LISTERIA used in combination with the CAMP test correctly identified 409 (98.8%) of 414 strains isolated from human, animal, food, and environmental sources belonging to the seven species currently defined within the genus Listeria. The kit was easy to use and simple to interpret. However, 8 of the 15 tests (i.e., phenylalanine deaminase, hydrogen sulfide, indole, ornithine decarboxylase, lysine decarboxylase, malonate, urease, and o-nitrophenyl-beta-D-galactopyranoside) were considered superfluous for the differentiation of Listeria spp. The CAMP test was indispensable when using the MICRO-ID LISTERIA system, in particular to differentiate CAMP test-positive L. monocytogenes from the nonhemolytic, rhamnose-positive L. innocua. The hemolytic L. seeligeri and L. ivanovii strains and the nonhemolytic, non-rhamnose-acidifying L. welshimeri strains could also be differentiated from one another only on the basis of their CAMP test results. The very few strains of L. grayi and L. murrayi were easily differentiated from the other nonhemolytic species. Catalase-negative cocci should not be tested, because 12 out of 19 catalase-negative strains (all enterococci) in our test were misidentified as Listeria spp. The MICRO-ID LISTERIA system identified strains within 18 to 24 h and is thus less time-consuming than conventional tests. The system could, therefore, be used together with correctly done CAMP tests for the rapid identification of Listeria isolates, especially food and environmental isolates, for which rapid species differentiation is important.
Protein-mediated adhesion of Lactobacillus acidophilus BG2FO4 on human enterocyte and mucus-secreting cell lines in culture
Authors: Coconnier M H , Klaenhammer T R , Kernéis S , Bernet M F , Servin A L
Published: January 2020

The adhesion of Lactobacillus acidophilus BG2FO4, a human stool isolate, to two human enterocytelike cell lines (Caco-2 and HT-29) and to the mucus secreted by a subpopulation of mucus-secreting HT29-MTX cells was investigated. Scanning electron microscopy revealed that the bacteria interacted with the well-defined apical microvilli of Caco-2 cells without cell damage and with the mucus secreted by the subpopulation of HT29-MTX cells. The adhesion to Caco-2 cells did not require calcium and involved an adhesion-promoting factor that was present in the spent supernatant of L. acidophilus cultures. This factor promoted adhesion of poorly adhering human Lactobacillus casei GG but did not promote adhesion of L. casei CNRZ 387, a strain of dairy origin. The adherence components on the bacterial cells and in the spent supernatant were partially characterized. Carbohydrates on the bacterial cell wall appeared to be partly responsible for the interaction between the bacteria and the extracellular adhesion-promoting factor. The adhesion-promoting factor was proteinaceous, since trypsin treatment dramatically decreased the adhesion of the L. acidophilus strain. The adhesion-promoting factor may be an important component of Lactobacillus species that colonize the gastrointestinal tract.
Glucose toxicity and inability of Bacteroides ruminicola to regulate glucose transport and utilization
Authors: Russell J B
Published: January 2020

Ammonia-limited (3.5 mM ammonia) cultures of Bacteroides ruminicola B(1)4 had a high number of viable cells (greater than 10(9)/ml), but only when the concentration of glucose was not too high (10 mM or less). When the glucose concentration was increased from 10 to 50 mM, there was a marked decrease in viability (10(5)-fold or greater). Because there was little decline in pH and only a small increase in succinate and acetate as the glucose concentration was increased, it did not appear that end products were killing the cells. This conclusion was supported by the observation that reinoculated cultures grew in the spent medium which had been supplemented with ammonia. Unlabeled rhamnose did not inhibit [14C]-glucose uptake, and cultures which were selected with a low concentration of rhamnose tolerated high concentrations of glucose (50 mM). The glucose-resistant mutant transported glucose at a lower rate than the wild type, and the Vmax of glucose transport was fourfold lower. The wild type stored much more polysaccharide than the glucose-resistant mutant, but it is not clear if polysaccharide accumulation per se is responsible for the glucose toxicity. These results indicated that B. ruminicola B(1)4 is unable to regulate glucose transport and utilization when growth is limited by ammonia.
Limited degradation of chlorophenols by anaerobic sludge granules
Authors: Mohn W W , Kennedy K J
Published: January 2020

To better understand the fate of chlorophenols treated in upflow anaerobic sludge bed reactors, we examined the ability of sludge granules from such bioreactors to degrade two trichlorophenols and one dichlorophenol in batch incubations under controlled conditions. Biodegradation was primarily limited to two distinct activities, reductive dehalogenation of ortho- and of meta-chlorine substituents. Both 3- and 4-monochlorophenol were persistent degradation products, while 2-monochlorophenol was further degraded. We also examined factors potentially affecting the rate and extent of 2,3,6-trichlorophenol degradation. An initial concentration of up to 1.75 mM (346 mg/liter) was dehalogenated. At that concentration, dehalogenation was partially inhibited but methanogenesis from formate was not. The initial concentration affected both the extent of dehalogenation and which products were detected. The maximum dechlorination rate observed was 1.4 mumol of Cl- h-1 g of volatile suspended solids-1. Dechlorination had a temperature optimum of 50 degrees C, was inhibited by added electron acceptors, and was not appreciably affected by added electron donors. The availability of electron acceptors and electron donors did not affect the extent of chlorophenol degradation. These particular sludge granules do not appear to be capable of mineralizing phenols with meta- or para-chlorine substituents.
Biodegradation of polycyclic aromatic hydrocarbons by new isolates of white rot fungi
Authors: Field J A , de Jong E , Feijoo Costa G , de Bont J A
Published: January 2020

Eight rapid Poly R-478 dye-decolorizing isolates from The Netherlands were screened in this study for the biodegradation of polycyclic aromatic hydrocarbons (PAH) supplied at 10 mg liter(-1). Several well-known ligninolytic culture collection strains, Phanerochaete chrysosporium BKM-F-1767, Trametes versicolor Paprican 52, and Bjerkandera adusta CBS 595.78 were tested in parallel. All of the strains significantly removed anthracene, and nine of the strains significantly removed benzo(a)pyrene beyond the limited losses observed in sterile liquid and HgCl2-poisoned fungus controls. One of the new isolates, Bjerkandera sp. strain Bos 55, was the best degrader of both anthracene and benzo(a)pyrene, removing 99.2 and 83.1% of these compounds after 28 days, respectively. Half of the strains, exemplified by strains of the genera Bjerkandera and Phanerochaete, converted anthracene to anthraquinone, which was found to be a dead-end metabolite, in high yields. The extracellular fluids of selected strains were shown to be implicated in this conversion. In contrast, four Trametes strains removed anthracene without significant accumulation of the quinone. The ability of Trametes strains to degrade anthraquinone was confirmed in this study. None of the strains accumulated PAH quinones during benzo(a)pyrene degradation. Biodegradation of PAH by the various strains was highly correlated to the rate by which they decolorized Poly R-478 dye, demonstrating that ligninolytic indicators are useful in screening for promising PAH-degrading white rot fungal strains.
Depletion of proton motive force by nisin in Listeria monocytogenes cells
Authors: Bruno M E , Kaiser A , Montville T J
Published: January 2020

The basal proton motive force (PMF) levels and the influence of the bacteriocin nisin on the PMF were determined in Listeria monocytogenes Scott A. In the absence of nisin, the interconversion of the pH gradient (Z delta pH) and the membrane potential (delta psi) led to the maintenance of a fairly constant PMF at -160 mV over the external pH range 5.5 to 7.0. The addition of nisin at concentrations of greater than or equal to 5 micrograms/ml completely dissipated PMF in cells at external pH values of 5.5 and 7.0. With 1 microgram of nisin per ml, delta pH was completely dissipated but delta psi decreased only slightly. The action of nisin on PMF in L. monocytogenes Scott A was both time and concentration dependent. Valinomycin depleted only delta pH, whereas nigericin and carbonyl cyanide m-chlorophenylhydrazone depleted only delta psi, under conditions in which nisin depleted both. Four other L. monocytogenes strains had basal PMF parameters similar to those of strain Scott A. Nisin (2.5 micrograms/ml) also completely dissipated PMF in these strains.
Roles of arginine in growth of Clostridium botulinum Okra B
Authors: Patterson-Curtis S I , Johnson E A
Published: January 2020

Group I strains of Clostridium botulinum are known to degrade arginine by the arginine deiminase pathway. We have found that C. botulinum Okra B consumed a level of arginine (20 g/liter) higher than the basal requirement for growth (3 g/liter). Arginine was probably the preferred source of nitrogen for biosynthesis but did not serve as a major source of energy. Citrulline and proline were produced as major fermentation products in media containing high levels of arginine, but in media with basal amounts of arginine these products were produced in lower quantities during growth and were later reassimilated. The results indicate that C. botulinum Okra B changes its metabolism during consumption of surplus arginine, and this change is associated with toxin repression, formation of citrulline and proline as end products, and possibly resistance to environmental stresses such as increased acidity and osmolarity.
Association of Luminous Bacteria with Artificial and Natural Surfaces in Arabian Gulf Seawater
Authors: Makemson John C. , Fulayfil Nada , Basson Philip
Published: January 2020

Luminous bacteria in seawater around the islands of Bahrain are predominantly
Vibrio harveyi
and have the capability to adhere to artificial fibrous surfaces. Phytoplankton did not appear to have any specific relationship with luminous bacteria, but macroalgae were shown to possess an enhanced concentration of luminous bacteria.
Molecular cloning of the Clostridium botulinum structural gene encoding the type B neurotoxin and determination of its entire nucleotide sequence
Authors: Whelan S M , Elmore M J , Bodsworth N J , Brehm J K , Atkinson T , Minton N P
Published: January 2020

DNA fragments derived from the Clostridium botulinum type A neurotoxin (BoNT/A) gene (botA) were used in DNA-DNA hybridization reactions to derive a restriction map of the region of the C. botulinum type B strain Danish chromosome encoding botB. As the one probe encoded part of the BoNT/A heavy (H) chain and the other encoded part of the light (L) chain, the position and orientation of botB relative to this map were established. The temperature at which hybridization occurred indicated that a higher degree of DNA homology occurred between the two genes in the H-chain-encoding region. By using the derived restriction map data, a 2.1-kb BglII-XbaI fragment encoding the entire BoNT/B L chain and 108 amino acids of the H chain was cloned and characterized by nucleotide sequencing. A contiguous 1.8-kb XbaI fragment encoding a further 623 amino acids of the H chain was also cloned. The 3' end of the gene was obtained by cloning a 1.6-kb fragment amplified from genomic DNA by inverse polymerase chain reaction. Translation of the nucleotide sequence derived from all three clones demonstrated that BoNT/B was composed of 1,291 amino acids. Comparative alignment of its sequence with all currently characterized BoNTs (A, C, D, and E) and tetanus toxin (TeTx) showed that a wide variation in percent homology occurred dependent on which component of the dichain was compared. Thus, the L chain of BoNT/B exhibits the greatest degree of homology (50% identity) with the TeTx L chain, whereas its H chain is most homologous (48% identity) with the BoNT/A H chain. Overall, the six neurotoxins were shown to be composed of highly conserved amino acid domains interceded with amino acid tracts exhibiting little overall similarity. In total, 68 amino acids of an average of 442 are absolutely conserved between L chains and 110 of 845 amino acids are conserved between H chains. Conservation of Trp residues (one in the L chain and nine in the H chain) was particularly striking. The most divergent region corresponds to the extreme carboxy terminus of each toxin, which may reflect differences in specificity of binding to neurone acceptor sites.
Degradation of azo dyes by the lignin-degrading fungus Phanerochaete chrysosporium
Authors: Spadaro J T , Gold M H , Renganathan V
Published: January 2020

Under nitrogen-limiting, secondary metabolic conditions, the white rot basidiomycete Phanerochaete chrysosporium extensively mineralized the specifically 14C-ring-labeled azo dyes 4-phenylazophenol, 4-phenylazo-2-methoxyphenol, Disperse Yellow 3 [2-(4'-acetamidophenylazo)-4-methylphenol], 4-phenylazoaniline, N,N-dimethyl-4-phenylazoaniline, Disperse Orange 3 [4-(4'-nitrophenylazo)-aniline], and Solvent Yellow 14 (1-phenylazo-2-naphthol). Twelve days after addition to cultures, the dyes had been mineralized 23.1 to 48.1%. Aromatic rings with substituents such as hydroxyl, amino, acetamido, or nitro functions were mineralized to a greater extent than unsubstituted rings. Most of the dyes were degraded extensively only under nitrogen-limiting, ligninolytic conditions. However, 4-phenylazo-[U-14C]phenol and 4-phenylazo-[U-14C]2-methoxyphenol were mineralized to a lesser extent under nitrogen-sufficient, nonligninolytic conditions as well. These results suggest that P. chrysosporium has potential applications for the cleanup of textile mill effluents and for the bioremediation of dye-contaminated soil.
Starvation Response of the Marine Barophile CNPT-3
Authors: Rice Scott A. , Oliver James D.
Published: January 2020

The psychrophilic marine barophile CNPT-3 underwent a starvation-survival response similar to that reported for the marine bacteria Ant-300, DW1, and S-14. The number of culturable cells increased initially and then decreased gradually over a 24-day starvation period, with corresponding decreases in total cell number and direct viability count. A significant reduction in cell size and biovolume accompanied these changes. Starved cells demonstrated a greater tendency to attach at the in situ pressure (400 atm; ca. 40.5 MPa) and temperature (5°C) than at 1 atm (ca. 101 kPa), and the extent of attachment increased with increasing duration of starvation. The membrane fatty acid profile of the marine barophile CNPT-3 was studied as the cells were subjected to starvation conditions. A 37.5% increase in saturated fatty acids was observed during the first 8 days of starvation, with a concomitant decrease in unsaturated fatty acids. There was also an increase in the amount of short-chain (<C
15:0
) fatty acids.
Rapid in situ hybridization technique using 16S rRNA segments for detecting and differentiating the closely related gram-positive organisms Bacillus polymyxa and Bacillus macerans
Authors: Jurtshuk R J , Blick M , Bresser J , Fox G E , Jurtshuk P
Published: January 2020

A rapid, sensitive, inexpensive in situ hybridization technique, using 30-mer 16S rRNA probes, can specifically differentiate two closely related Bacillus spp., B. polymyxa and B. macerans. The 16S rRNA probes were labeled with a rhodamine derivative (Texas Red), and quantitative fluorescence measurements were made on individual bacterial cells. The microscopic fields analyzed were selected by phase-contrast microscopy, and the fluorescence imaging analyses were performed on 16 to 67 individual cells. The labeled 16S rRNA probe, POL, whose sequence was a 100% match with B. polymyxa 16S rRNA but only a 60% match with B. macerans 16S rRNA, gave quantitative fluorescence ratio measurements that were 34.8-fold higher for B. polymyxa cells than for B. macerans cells. Conversely, the labeled probe, MAC, which matched B. polymyxa 16S rRNA in 86.6% of its positions and B. macerans 16S rRNA in 100% of its positions, gave quantitative fluorescence measurements that were 59.3-fold higher in B. macerans cells than in B. polymyxa cells. Control probes, whose 16S rRNA sequence segment (P-M) was present in both B. polymyxa and B. macerans as well as a panprokaryotic probe (16S), having a 100% match with all known bacteria, hybridized equally well with both organisms. These latter hybridizations generated very high fluorescence signals, but their comparative fluorescence ratios (the differences between two organisms) were low. The control paneukaryotic probe (28S), which had less than 30% identity for both B. macerans and B. polymyxa, did not hybridize with either organism.
Contribution of phenazine antibiotic biosynthesis to the ecological competence of fluorescent pseudomonads in soil habitats
Authors: Mazzola M , Cook R J , Thomashow L S , Weller D M , Pierson L S
Published: January 2020

Phenazine antibiotics produced by Pseudomonas fluorescens 2-79 and Pseudomonas aureofaciens 30-84, previously shown to be the principal factors enabling these bacteria to suppress take-all of wheat caused by Gaeumannomyces graminis var. tritici, also contribute to the ecological competence of these strains in soil and in the rhizosphere of wheat. Strains 2-79 and 30-84, their Tn5 mutants defective in phenazine production (Phz-), or the mutant strains genetically restored for phenazine production (Phz+) were introduced into Thatuna silt loam (TSL) or TSL amended with G. graminis var. tritici. Soils were planted with three or five successive 20-day plant-harvest cycles of wheat. Population sizes of Phz- derivatives declined more rapidly than did population sizes of the corresponding parental or restored Phz+ strains. Antibiotic biosynthesis was particularly critical to survival of these strains during the fourth and fifth cycles of wheat in the presence of G. graminis var. tritici and during all five cycles of wheat in the absence of take-all. In pasteurized TSL, a Phz- derivative of strain 30-84 colonized the rhizosphere of wheat to the same extent that the parental strain did. The results indicate that production of phenazine antibiotics by strains 2-79 and 30-84 can contribute to the ecological competence of these strains and that the reduced survival of the Phz- strains is due to a diminished ability to compete with the resident microflora.
Use of degenerate primers for polymerase chain reaction cloning and sequencing of the Lactococcus lactis subsp. lactis recA gene
Authors: Duwat P , Ehrlich S D , Gruss A
Published: January 2020

Two particularly well-conserved stretches in the RecA protein sequences were chosen as templates to synthesize degenerate oligonucleotides, which were used in polymerase chain reaction to amplify an internal recA DNA fragment of Lactococcus lactis subsp. lactis ML3. Using this fragment, we recovered and sequenced the entire lactococcal recA gene. The end of an open reading frame present upstream of the recA gene shows strong homology with formamidopyrimidine-DNA-glycosylase, a protein involved in DNA repair.
Genetic analysis of a locus on the Bacteroides ovatus chromosome which contains xylan utilization genes
Authors: Weaver J , Whitehead T R , Cotta M A , Valentine P C , Salyers A A
Published: January 2020

Bacteroides ovatus, a gram-negative obligate anaerobe found in the human colon, can utilize xylan as a sole source of carbohydrate. Previously, a 3.8-kbp segment of B. ovatus chromosomal DNA, which contained genes encoding a xylanase (xylI) and a bifunctional xylosidase-arabinosidase (xsa), was cloned, and expression of the two genes was studied in Escherichia coli (T. Whitehead and R. Hespell, J. Bacteriol. 172:2408-2412, 1990). In the present study, we have used segments of the cloned region to construct insertional disruptions in the B. ovatus chromosomal locus containing these two genes. Analysis of these insertional mutants demonstrated that (i) xylI and xsa are probably part of the same operon, with xylI upstream of xsa, (ii) the true B. ovatus promoter was not cloned on the 3.5-kbp DNA fragment which expressed xylanase and xylosidase in E. coli, (iii) there is at least one gene upstream of xylI which could encode an arabinosidase, and (iv) xylosidase rather than xylanase may be a rate-limiting step in xylan utilization. Insertional mutations in the xylI-xsa locus reduced the rate of growth on xylan, but the concentration of residual sugars at the end of growth was the same as that with the wild type. Thus, a slower rate of growth on xylan was not accompanied by less extensive digestion of xylan. Mutants in which xylI had been disrupted still expressed some xylanase activity. This second activity was associated with membranes and produced xylose from xylan, whereas the xylI gene product partitioned primarily with the soluble fraction and produced xylobiose from xylan.
Induction of melanin biosynthesis in Vibrio cholerae
Authors: Coyne V E , al-Harthi L
Published: January 2020

Vibrio cholerae synthesized the pigment melanin in response to specific physiological conditions that were stressful to the bacterium. Pigmentation was induced when V. cholerae was subjected to hyperosmotic stress in conjunction with elevated growth temperatures (above 30 degrees C). The salt concentration tolerated by V. cholerae was lowered by additional abiotic factors such as acidic starting pH of the growth medium and limitation of organic nutrients. Although the amount of toxin detected in the culture supernatant decreased significantly in response to stressful culture conditions, no correlation between the physiological conditions that induced melanogenesis and expression of OmpU or cholera toxin was detected. Since conditions that induce melanin production in V. cholerae occur in both the aquatic environment and the human host, it is possible that melanogenesis has a specific function with respect to the survival of the bacterium in these habitats.
Degradation of phenanthrene by Phanerochaete chrysosporium occurs under ligninolytic as well as nonligninolytic conditions
Authors: Dhawale S W , Dhawale S S , Dean-Ross D
Published: January 2020

In order to delineate the roles of lignin and manganese peroxidases in the degradation of polycyclic aromatic hydrocarbons by Phanerochaete chrysosporium, the biodegradation of phenanthrene (chosen as a model for polycyclic aromatic hydrocarbons) was investigated. The disappearance of phenanthrene from the extracellular medium and mycelia was determined by using gas chromatography. The disappearance of phenanthrene from cultures of wild-type strains BKM-F1767 (ATCC 24725) and ME446 (ATCC 34541) under ligninolytic (low-nitrogen) as well as nonligninolytic (high-nitrogen) conditions was observed. The study was extended to two homokaryotic (basidiospore-derived) isolates of strain ME446. Both homokaryotic isolates, ME446-B19 (which produces lignin and manganese peroxidases only in low-nitrogen medium) and ME446-B5 (which totally lacks lignin and manganese peroxidase activities), caused the disappearance of phenanthrene when grown in low- as well as high-nitrogen media. Moreover, lignin and manganese peroxidase activities were not detected in any of the cultures incubated in the presence of phenanthrene. Additionally, the mineralization of phenanthrene was observed even under nonligninolytic conditions. The results collectively indicate that lignin and manganese peroxidases are not essential for the degradation of phenanthrene by P. chrysosporium. The observation that phenanthrene degradation occurs under nonligninolytic conditions suggests that the potential of P. chrysosporium for degradation of certain environmental pollutants is not limited to nutrient starvation conditions.
Genomic analysis of Pediococcus starter cultures used to control Listeria monocytogenes in turkey summer sausage
Authors: Luchansky J B , Glass K A , Harsono K D , Degnan A J , Faith N G , Cauvin B , Baccus-Taylor G , Arihara K , Bater B , Maurer A J
Published: January 2020

The pulsed-field technique of clamped homogeneous electric field electrophoresis was employed to characterize and size genomic DNA of three pediocin-producing (Ped+) and two non-pediocin-producing (Ped-) strains of Pediococcus acidilactici. Comparison of genomic fingerprints obtained by digestion with the low-frequency-cleavage endonuclease AscI revealed identical restriction profiles for four of the five strains analyzed. Summation of results for 10 individually sized AscI fragments estimated the genome length to be 1,861 kb for the four strains (H, PAC1.0, PO2, and JBL1350) with identical fingerprints. Genomic analysis of the pediocin-sensitive, plasmid-free strain P. acidilactici LB42 with the unique fingerprint revealed nine AscI fragments and a genome length of about 2,133 kb. Ped- (JBL1350) and Ped+ (JBL1095) starter cultures (one each) were used to separately prepare turkey summer sausage coinoculated with a four-strain Listeria monocytogenes mixture (ca. 10(5) CFU/g). The starter cultures produced equivalent amounts of acid during fermentation, but counts of L. monocytogenes were reduced to a greater extent in the presence of the Ped+ starter culture (3.4 log10 unit decrease) than in the presence of the Ped- starter culture (0.9 log10 unit decrease). Although no listeriae were recovered from sausages following the cook/shower, appreciable pediocin activity was recovered from sausages prepared with the Ped+ strain for at least 60 days during storage at 4 degrees C. The results of this study revealed genomic similarities among pediococcal starter cultures and established that pediocins produced during fermentation provide an additional measure of safety against listerial proliferation in turkey summer sausage.
Rates of inactivation of waterborne coliphages by monochloramine
Authors: Dee S W , Fogleman J C
Published: January 2020

A sophisticated water quality monitoring program was established to evaluate virus removal through Denver's 1-million-gal (ca. 4-million-liter)/day Direct Potable Reuse Demonstration Plant. As a comparison point for the reuse demonstration plant, Denver's main water treatment facility was also monitored for coliphage organisms. Through the routine monitoring of the main plant, it was discovered that coliphage organisms were escaping the water treatment processes. Monochloramine residuals and contact times (CT values) required to achieve 99% inactivation were determined for coliphage organisms entering and leaving this conventional water treatment plant. The coliphage tested in the effluent waters had higher CT values on the average than those of the influent waters. CT values established for some of these coliphages suggest that monochloramine alone is not capable of removing 2 orders of magnitude of these specific organisms in a typical water treatment facility. Electron micrographs revealed one distinct type of phage capable of escaping the water treatment processes and three distinct types of phages in all.
Identification of geosmin as a volatile metabolite of Penicillium expansum
Authors: Mattheis J P , Roberts R G
Published: January 2020

Cultures of Penicillium expansum produce a musty, earthy odor. Geosmin [1,10-trans-dimethyl-trans(9)-decalol] was identified by gas chromatography-mass spectrometry from headspace samples of P. expansum cultures. Olfactory comparison of P. expansum cultures with a geosmin standard indicated geosmin is the primary component of the odor associated with P. expansum.
Effect of Endosulfan on
Azospirillum lipoferum
Growth, Morphology, Nitrogenase Activity, and Protein Binding
Authors: Buff Klaus , Mano Denise M. S. , Langenbach Tomaz
Published: January 2020

The organochlorine Thiodan CE inhibited growth and nitrogenase activity of
Azospirillum lipoferum.
The active ingredient, Endosulfan, was nonspecifically bound to proteins and mainly adsorbed to the cell envelope with small amounts transported into cytosol. The involvement of the external membrane and cyst formation in protection against hazardous substances is discussed.
Comparison of Selected Methods for the Enumeration of Fecal Coliforms and
Escherichia coli
in Shellfish
Authors: Grabow W. O. K. , De Villiers J. C. , Schildhauer C. I.
Published: January 2020

In a comparison of five selected methods for the enumeration of fecal coliforms and
Escherichia coli
in naturally contaminated and sewage-seeded mussels (
Choromytilus
spp.) and oysters (
Ostrea
spp.), a spread-plate procedure with mFC agar without rosolic acid and preincubation proved the method of choice for routine quality assessment.
Characterization of a Cell Envelope-Associated Proteinase Activity from
Streptococcus thermophilus
H-Strains
Authors: Shahbal Samaha , Hemme Denis , Renault Pierre
Published: January 2020

The production and biochemical properties of cell envelope-associated proteinases from two strains of
Streptococcus thermophilus
(strains CNRZ 385 and CNRZ 703) were compared. No significant difference in proteinase activity was found for strain CNRZ 385 when cells were grown in skim milk medium and M17 broth. Strain CNRZ 703 exhibited a threefold-higher proteinase activity when cells were grown in low-heat skim milk medium than when grown in M17 broth. Forty-one percent of the total activity of CNRZ 385 was localized on the cell wall. The optimum pH for enzymatic activity at 37°C was around 7.0. Serine proteinase inhibitors, such as phenylmethylsulfonyl fluoride and diisopropylfluorophosphate, inhibited the enzyme activity in both strains. The divalents cations Ca
2+
, Mg
2+
, and Mn
2+
were activators, while Zn
2+
and Cu
2+
were inhibitors. β-Casein was hydrolyzed more rapidly than α
s1
-casein. The results of DNA hybridization and immunoblot studies suggested that the
S. thermophilus
cell wall proteinase and the lactococcal proteinase are not closely related.
Inhibition of adhesion of Escherichia coli K88 to piglet ileal mucus by Lactobacillus spp
Authors: Blomberg L , Henriksson A , Conway P L
Published: January 2020

Enteropathogenic Escherichia coli K88 colonizing the piglet ileum adhere to the mucosa by K88 fimbrial appendages. A recent study in our laboratory has implicated indigenous lactobacilli in the suppression of the colonization potential of enteropathogenic E. coli as measured by adhesion to ileal mucus. The aim of this study was to investigate the effect of Lactobacillus spp. of porcine origin on the adhesion of K88 fimbriae of E. coli. With an in vitro assay, the adhesion of E. coli K88ab strain G1108E and E. coli K88ac strain 1107 to 35-day-old piglet ileal mucus was studied in the presence of spent culture fluid of Lactobacillus spp. Detailed studies focused specifically on culture fluid of Lactobacillus fermentum 104R. Subsequently, the ileal mucus was exposed to the retentate of the spent culture fluid after dialysis and fractionation. Adhesion was confirmed to be attributable to K88 fimbriae when K88-specific monoclonal antibodies and isogenic mutants of E. coli K-12 with and without the plasmid containing the K88 gene were used. The active component was characterized by pretreatment of dialysis retentate with heat, periodate, pronase, and centrifugation, as well as by growth of the lactobacillus in various media and by assays at both 0 and 37 degrees C. All three lactobacilli of porcine origin reduced adhesion of E. coli K88 by approximately 50%. Inhibition occurred when mucus was pretreated with either spent culture dialysis retentate or the void volume (fraction of > 250,000 molecular weight) after gel filtration. The activity of the dialysis retentate was sensitive to pronase, but there was still activity at 0 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)
Pediocin PA-1, a bacteriocin from Pediococcus acidilactici PAC1.0, forms hydrophilic pores in the cytoplasmic membrane of target cells
Authors: Chikindas M L , García-Garcerá M J , Driessen A J , Ledeboer A M , Nissen-Meyer J , Nes I F , Abee T , Konings W N , Venema G
Published: January 2020

Pediocin PA-1 is a bacteriocin which is produced by Pediococcus acidilactici PAC1.0. We demonstrate that pediocin PA-1 kills sensitive Pediococcus cells and acts on the cytoplasmic membrane. In contrast to its lack of impact on immune cells, pediocin PA-1 dissipates the transmembrane electrical potential and inhibits amino acid transport in sensitive cells. Pediocin interferes with the uptake of amino acids by cytoplasmic membrane vesicles derived from sensitive cells, while it is less effective with membranes derived from immune cells. In liposomes fused with membrane vesicles derived from both sensitive and immune cells, pediocin PA-1 elicits an efflux of small ions and, at higher concentrations, an efflux of molecules having molecular weights of up to 9,400. Our data suggest that pediocin PA-1 functions in a voltage-independent manner but requires a specific protein in the target membrane.
Isolation and Molecular Characterization of Five Marine Cyanophages Propagated on
Synechococcus
sp. Strain WH7803
Authors: Wilson William H. , Joint Ian R. , Carr Noel G. , Mann Nicholas H.
Published: January 2020

Five marine cyanophages propagated on
Synechococcus
sp. strain WH7803 were isolated from three different oceanographic provinces during the months of August and September 1992: coastal water from the Sargasso Sea, Bermuda; Woods Hole harbor, Woods Hole, Mass.; and coastal water from the English Channel, off Plymouth Sound, United Kingdom. The five cyanophage isolates were found to belong to two families, Myoviridae and Styloviridae, on the basis of their morphology observed in the transmission electron microscope. DNA purified from each of the cyanophage isolates was restricted with a selection of restriction endonucleases, and three distinguishably different patterns were observed. DNA isolated from Myoviridae isolates from Bermuda and the English Channel had highly related restriction patterns, as did DNA isolated from Styloviridae isolates from Bermuda and the English Channel. DNA isolated from the Myoviridae isolate from Woods Hole had a unique restriction pattern. The genome size for each of the Myoviridae isolates was ca. 80 to 85 kb, and it was ca. 90 to 100 kb for each of the Styloviridae isolates. Southern blotting analysis revealed that there was a limited degree of homology among all cyanophage DNAs probed, but clear differences were observed between cyanophage DNA from the Myoviridae and that from the Styloviridae isolates. Polypeptide analysis revealed a clear difference between Myoviridae and Styloviridae polypeptide profiles, although the major, presumably structural, protein in each case was ca. 53 to 54 kDa.
Electrochemical Prevention of Marine Biofouling with a Carbon-Chloroprene Sheet
Authors: Nakasono Satoshi , Burgess J. Grant , Takahashi Kanjo , Koike Masami , Murayama Chisei , Nakamura Shigeoki , Matsunaga Tadashi
Published: January 2020

A carbon-chloroprene sheet (CCS) electrode was used for the electrochemical disinfection of the marine gram-negative bacterium
Vibrio alginolyticus
. When the electrode was incubated in seawater containing 10
5
cells per ml for 90 min, the amount of adsorbed cells was 4.5 × 10
3
cells per cm
2
. When a potential of 1.2 V versus a saturated calomel electrode was applied to the CCS for 20 min, 67% of adsorbed cells were killed. This disinfection was due to the direct electrochemical oxidation of cells and not to a change in pH or to the generation of toxic substances, such as chlorine. In a 1-year field experiment, marine biofouling of a CCS-coated cooling pipe caused by attachment of bacteria and invertebrates was considerably reduced by application of a potential of 1.2 V versus a saturated calomel electrode. Since this method requires low potential electrical energy, use of a CCS coating appears to be a suitable method for the clean prevention of marine biofouling.
Disaggregation of
Methanosarcina
spp. and Growth as Single Cells at Elevated Osmolarity
Authors: Sowers Kevin R. , Boone Jane E. , Gunsalus Robert P.
Published: January 2020

The effect of medium osmolarity on the morphology and growth of
Methanosarcina barkeri, Methanosarcina thermophila, Methanosarcina mazei, Methanosarcina vacuolata,
and
Methanosarcina acetivorans
was examined. Each strain was adapted for growth in NaCl concentrations ranging from 0.05 to 1.0 M.
Methanosarcina
spp. isolated from both marine and nonmarine sources exhibited similar growth characteristics at all NaCl concentrations tested, demonstrating that these species are capable of adapting to a similar range of medium osmolarities. Concomitant with the adaptation in 0.4 to 1.0 M NaCl, all strains disaggregated and grew as single cells rather than in the characteristic multicellular aggregates. Aggregated cells had a methanochondroitin outer layer, while disaggregated single cells lacked the outer layer but retained the protein S-layer adjacent to the cell membrane. Synthesis of glucuronic acid, a major component of methanochondroitin, was reduced 20-fold in the single-cell form of
M. barkeri
when compared with synthesis in aggregated cells. Strains with the methanochondroitin outer cell layer exhibited enhanced stability at low (<0.2 M NaCl) osmolarity and grew at higher temperatures. Disaggregated cells could be converted back to aggregated cells by gradually readapting cultures to lower NaCl (<0.2 M) and Mg
2+
(<0.005 M) concentrations. Disaggregated
Methanosarcina
spp. could also be colonized and replica plated with greater than 95% recovery rates on solidified agar basal medium that contained 0.4 to 0.6 M NaCl and either trimethylamine, methanol, or acetate as the substrate. The ability to disaggregate and grow
Methanosarcina
spp. as viable, detergent-sensitive, single cells on agar medium makes these species amenable to mutant selection and screening for genetic studies and enables cells to be gently lysed for the isolation of intact genetic material.
Presence of Tetrodotoxin and Tetrodotoxin-Producing Bacteria in Freshwater Sediments
Authors: Do H. K. , Hamasaki K. , Ohwada K. , Simidu U. , Noguchi T. , Shida Y. , Kogure K.
Published: January 2020

The occurrence of tetrodotoxin (TTX) in sediment from Lake Suwa, Japan, was confirmed by a tissue culture assay, high-performance liquid chromatography, and gas chromatography-mass spectrometry. Altogether, 17 TTX-producing bacteria belonging to five genera were isolated from Lake Suwa and Pond Inokasira. Our results indicate that TTX and TTX-producing bacteria occur even in freshwater environments.