bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of media for monitoring fecal streptococci in seawater
Authors: Yoshpe-Purer Y
Published: January 2020

The selectivity of KF streptococcus agar (KF) for monitoring fecal streptococci (FS) in seawater was examined in 234 samples of Mediterranean water and compared with the selectivity of M-Enterococcus agar (M-Ent) for 124 samples and with bile-esculin-azide agar (BEA) for 17 samples. KF was found to be unsuitable for marine water because Vibrio alginolyticus and other gram-negative bacilli indigenous to this environment grew well on it and produced red colonies identical to those of FS. In 26% of samples, some with high counts of red colonies on the membrane filters (MF), there were no streptococci, only gram-negative bacilli and staphylococci, and in an additional 23.1% the streptococci constituted less than 50% of the "typical" red colonies on the MF. V. alginolyticus also produced FS-like colonies on MF incubated on BEA but was not isolated from MF incubated on M-Ent. Although staphylococci grew and produced FS-like colonies on all three media, M-Ent was the most selective since no gram-negative bacilli were isolated from MF incubated on it.
Localization of Bacteria and Hemoglobin in Root Nodules of
Parasponia andersonii
Containing Both
Bradyrhizobium
Strains and
Rhizobium leguminosarum
biovar
trifolii
Authors: Trinick Michael J. , Goodchild David J. , Miller Celia
Published: January 2020

Dual occupancy of
Parasponia andersonii
nodules with different
Bradyrhizobium
strains and
Rhizobium leguminosarum
biovar
trifolii
was frequently obtained when two strains were inoculated into plants grown aseptically in tubes. Since reisolates of
Bradyrhizobium
strains from dually occupied nodules acquired the ability to nodulate
Trifolium repens
, the spatial relationship of the two species of bacteria during nodule initiation and development was investigated and their proximity was demonstrated. By using light microscopy and electron microscopy and immunogold labeling,
R. leguminosarum
biovar
trifolii
NGR66 inoculated alone onto
P. andersonii
produced small ineffective nodules, with bacteria embedded in matrix material in intercellular spaces and in a few nonliving host cells rather than in infection threads (CP299). In dual infections, the two bacterial species were shown to be adjacent to one another in the matrix of nodule intercellular spaces and in some host nodule cells. However, when two different
Bradyrhizobium
strains occupied a single nodule, they were located in different lobes of the same nodule. Immunogold labeling showed that
Parasponia
hemoglobin was localized in the cytoplasm of young infected nodule cells. This suggests that the nitrogen-fixing phase of
Parasponia
nodule cells is short-lived and correlates with previous acetylene reduction data from nodule slices. Hemoglobin was associated only with areas of nodule tissue infected with the effective nitrogen-fixing strain CP299 and absent from areas infected with
R. leguminosarum
biovar
trifolii
.
Surface-active properties of Candida albicans
Authors: Klotz S A
Published: January 2020

Cell surface hydrophobicity may be an important factor contributing to the virulence of Candida yeast cells. Surface hydrophobic and surface polar groups would be required for a yeast cell to act as a surface-active agent. In this report, the surface activities of whole yeast cells were measured. Yeast cells added at 10(8)/ml reduced the surface tension (gamma s) of saline by 20% as determined by the du Nouy method. A 1% suspension of yeast cell wall fragments reduced gamma s of saline by 36%. Whole yeast cells caused a reduction in interfacial tension (gamma I) between hexadecane and saline. The reduction of gamma I was proportional to the surface hydrophobicity of the yeasts. Yeast cells grown in glucose as the sole carbon source (thus possessing a relatively more hydrophilic cell surface) reduced gamma I by 30%, whereas yeast cells grown in hexadecane (thus possessing a more hydrophobic cell surface) reduced gamma I by 41%. The reduction of gamma I was reversed upon the addition of a strong surfactant. It was also demonstrated that yeast cells blended with nonionic surfactants during growth in a glucose broth in order to change their cell surface hydrophobicity adhered to solid surfaces in direct proportion to their cell surface hydrophobicity. Thus, the surface-active properties of Candida yeast cells may significantly contribute to the accumulation of yeast cells at various biological interfaces such as liquid-solid, liquid-liquid, and liquid-air, leading to their eventual adhesion to solid or tissue surfaces.
Mass-spectrometric studies of the interrelations among hydrogenase, carbon monoxide dehydrogenase, and methane-forming activities in pure and mixed cultures of Desulfovibrio vulgaris, Desulfovibrio desulfuricans, and Methanosarcina barkeri
Authors: Rajagopal B S , Lespinat P A , Fauque G , LeGall J , Berlier Y M
Published: January 2020

The activities of pure and mixed cultures of Desulfovibrio vulgaris and Methanosarcina barkeri in the exponential growth phase were monitored by measuring changes in dissolved-gas concentration by membrane-inlet mass spectrometry. M. barkeri grown under H2-CO2 or methanol produced limited amounts of methane and practically no hydrogen from either substrate. The addition of CO resulted in a transient H2 production concomitant with CO consumption. Hydrogen was then taken up, and CH4 production increased. All these events were suppressed by KCN, which inhibited carbon monoxide dehydrogenase activity. Therefore, with both substrates, H2 appeared to be an intermediate in CO reduction to CH4. The cells grown on H2-CO2 consumed 4 mol of CO and produced 1 mol of CH4. Methanol-grown cells reduced CH3OH with H2 resulting from carbon monoxide dehydrogenase activity, and the ratio was then 1 mol of CH4 to 1 mol of CO. Only 12CH4 and no 13CH4 was obtained from 13CO, indicating that CO could not be the direct precursor of CH4. In mixed cultures of D. vulgaris and M. barkeri on lactate, an initial burst of H2 was observed, followed by a lower level of production, whereas methane synthesis was linear with time. Addition of CO to the mixed culture also resulted in transient extra H2 production but had no inhibitory effect upon CH4 formation, even when the sulfate reducer was D. vulgaris Hildenborough, whose periplasmic iron hydrogenase is very sensitive to CO. The hydrogen transfer is therefore probably mediated by a less CO-sensitive nickel-iron hydrogenase from either of both species.
Integration and expression of alpha-amylase and endoglucanase genes in the Lactobacillus plantarum chromosome
Authors: Scheirlinck T , Mahillon J , Joos H , Dhaese P , Michiels F
Published: January 2020

A commercial grass silage starter strain of Lactobacillus plantarum was transformed by high-frequency electroporation with plasmids containing an alpha-amylase gene from Bacillus stearothermophilus and an endoglucanase gene from Clostridium thermocellum. Both genes were expressed from their native regulatory signals, and active enzymes were found in the supernatant. However, the segregational stability of the transforming plasmids was rather low. Therefore, the transforming genes were inserted in the L. plantarum chromosome by means of single homologous recombination. In the majority of the transformants, this led to extremely stable segregation and expression of the transforming genes, without generating secondary mutations in the host. Increased selective pressure led to tandem amplification of the transforming DNA. The transformed strains demonstrated the ability of L. plantarum to express heterologous gene products; they can be used to detect the inoculum in silage ecology studies; and they demonstrate the feasibility of engineering truly cellulolytic silage starter bacteria.
Biological reductive dechlorination of tetrachloroethylene and trichloroethylene to ethylene under methanogenic conditions
Authors: Freedman D L , Gossett J M
Published: January 2020

A biological process for remediation of groundwater contaminated with tetrachloroethylene (PCE) and trichloroethylene (TCE) can only be applied if the transformation products are environmentally acceptable. Studies with enrichment cultures of PCE- and TCE-degrading microorganisms provide evidence that, under methanogenic conditions, mixed cultures are able to completely dechlorinate PCE and TCE to ethylene, a product which is environmentally acceptable. Radiotracer studies with [14C]PCE indicated that [14C]ethylene was the terminal product; significant conversion to 14CO2 or 14CH4 was not observed. The rate-limiting step in the pathway appeared to be conversion of vinyl chloride to ethylene. To sustain reductive dechlorination of PCE and TCE, it was necessary to supply an electron donor; methanol was the most effective, although hydrogen, formate, acetate, and glucose also served. Studies with the inhibitor 2-bromoethanesulfonate suggested that methanogens played a key role in the observed biotransformations of PCE and TCE.
Two distinct O-methyltransferases in aflatoxin biosynthesis
Authors: Yabe K , Ando Y , Hashimoto J , Hamasaki T
Published: January 2020

The substances belonging to the sterigmatocystin group bear a close structural relationship to aflatoxins. When demethylsterigmatocystin (DMST) was fed to Aspergillus parasiticus NIAH-26, which endogenously produces neither aflatoxins nor precursors in YES medium, aflatoxins B1 and G1 were produced. When dihydrodemethylsterigmatocystin (DHDMST) was fed to this mutant, aflatoxins B2 and G2 were produced. Results of the cell-free experiment with S-adenosyl-[methyl-3H]methionine showed that first the C-6-OH groups of DMST and DHDMST are methylated to produce sterigmatocystin and dihydrosterigmatocystin (O-methyltransferase I) and then the C-7-OH groups are methylated to produce O-methylsterigmatocystin (OMST) and dihydro-O-methylsterigmatocystin (DHOMST) (O-methyltransferase II). However, no methyltransferase activity was observed when either OMST, DHOMST, 5,6-dimethoxysterigmatocystin, 5-methoxysterigmatocystin, or sterigmatin was incubated with the cell extract. Treatment of the cell extract with N-ethylmaleimide inhibited O-methyltransferase I activity but not that of O-methyltransferase II. Furthermore, these O-methyltransferases were different in their protein molecules and were involved in both the reactions from DMST to OMST and DHDMST to DHOMST. The reactions described in this paper were not observed when the same mold had been cultured in YEP medium.
The cutworm Peridroma saucia (Lepidoptera: Noctuidae) supports growth and transport of pBR322-bearing bacteria
Authors: Armstrong J L , Porteous L A , Wood N D
Published: January 2020

Variegated cutworms were exposed to bean plants in microcosms sprayed with pBR322-carrying strains of Enterobacter cloacae, Klebsiella planticola, and Erwinia herbicola. The three bacterial species exhibited differential survival on leaves, in soil, and in guts and fecal pellets (frass) of the insects. High numbers of Enterobacter cloacae(pBR322) were detected in all samples, while the other species were unable to establish residence in the insect. To assess the impact of this colonization on site-to-site transport of microorganisms, larvae were fed plants that had been sprayed with the bacteria and then were transferred to uninoculated plants. Cutworms were efficient carriers of Enterobacter cloacae(pBR322), as indicated by its rapid appearance on uninoculated leaves and continued persistence in the insects for 3 days after transfer. Few Erwinia herbicola(pBR322) and K. planticola(pBR322) were obtained from larvae after transfer, although up to 10(3) CFU/g were detected in soil and on plants. Differences in bacterial survival and growth were confirmed by incubating frass overnight and observing the change in population numbers. The proportion of total samples showing at least a 25-fold increase during incubation was 68% for Enterobacter cloacae(pBR322), 39% for K. planticola(pBR322), and 0% for Erwinia herbicola(pBR322). Our results emphasize the role that cutworms and possibly other insects have in persistence and growth of microorganisms in the environment.
Transformation of Saccharomyces cerevisiae by electroporation
Authors: Delorme E
Published: January 2020

A method for introducing heterologous DNA into Saccharomyces cerevisiae rapidly and efficiently by electroporation was developed. Transformant colonies appeared somewhat sooner than by the LiCl or spheroplast transformation method, and the time spent in manipulation was much less than for these two methods. The pores in the cell membrane formed by the high voltage of electroporation were resealed within 6 to 7 min after electroporation. At a capacitance of 25 microF, the optimum voltage was 2.0 to 2.25 kV/cm. Log-phase cells concentrated to 10 to 20 units at an optical density of 600 nm in 200 microliters of fresh rich medium and electroporated at 2.25 kV/cm in the presence of 0.1 microgram of supercoiled plasmid DNA will yield 1,000 to 4,500 colonies per microgram of DNA.
Acetate Production by Methanogenic Bacteria
Authors: Westermann Peter , Ahring Birgitte K. , Mah Robert A.
Published: January 2020

Methanosarcina barkeri
MS and 227 and
Methanosarcina mazei
S-6 produced acetate when grown on H
2
-CO
2
, methanol, or trimethylamine. Marked differences in acetate production by the two bacterial species were found, even though methane and cell yields were nearly the same.
M. barkeri
produced 30 to 75 μmol of acetate per mmol of CH
4
formed, but
M. mazei
produced only 8 to 9 μmol of acetate per mmol of CH
4
.
Preliminary characterization of a food-borne multiple-antibiotic-resistant Salmonella typhimurium strain
Authors: Schuman J D , Zottola E A , Harlander S K
Published: January 2020

Plasmid characterization studies were conducted on a Salmonella typhimurium strain isolated from pasteurized milk and from a symptomatic patient during the 1985 Illinois salmonellosis outbreak. This strain (Hf) was reported to possess an unusual plasmid profile which distinguished it from all Salmonella strains isolated in the United States prior to 1984. Antibiotic susceptibility testing revealed that the strain was resistant to tetracycline, erythromycin, clindamycin, sulfisoxazole, sulfadiazene, triple sulfa, cefoperazone, streptomycin, mezlocillin, piperacillin, carbenicillin, penicillin, ampicillin, and kanamycin. Plasmid analysis revealed that the strain possessed four plasmids with sizes of approximately 158, 98, 10.2, and 6.0 kilobase pairs (kb). Successive transfer at 43 degrees C led to increased antibiotic sensitivity in 75.5% of the isolates screened. Electroporation and calcium chloride treatment were each used to transform plasmid-free Escherichia coli strains with the plasmid pool from S. typhimurium Hf. Plasmids introduced by transformation ranged in size from 4.4 to 23.2 kb and correlated with resistance to penicillin G, ampicillin, carbenicillin, cephalothin, cefoperazone, cefamandole, mezlocillin, piperacillin, and in some cases, tetracycline and kanamycin. DNA-DNA hybridization experiments localized these resistance genes to a highly duplicated 6.3-kb fragment of the total EcoRI restriction digest of the S. typhimurium Hf plasmid pool.
Detection, Growth, and Amine-Producing Capacity of Lactobacilli in Cheese
Authors: Joosten H. M. L. J. , Northolt M. D.
Published: January 2020

A differential plating medium was developed to detect decarboxylating lactobacilli in cheese. With this medium, 15 cheeses made from raw milk were investigated for the presence of these bacteria. Five histidine-decarboxylating strains and one tyrosine-decarboxylating strain were isolated. The isolates were identified with the API 50L system. Accordingly, each of the five histidine-decarboxylating strains was identified as
Lactobacillus buchneri
, whereas the tyrosine-decarboxylating strain is a representative of
Lactobacillus brevis.
Cheesemaking experiments using a low inoculum concentration of the histidinedecarboxylating
L. buchneri
strain St2A (0.2 CFU/ml of milk) showed that, under conditions of accelerated proteolysis, histamine may accumulate rapidly; after 3 months of ripening, 410 mg/kg was found. An inoculum concentration of 5 CFU/ml gave rise to the formation of 1,060 mg/kg.
Analysis of peptide metabolism by ruminal microorganisms
Authors: Wallace R J , McKain N
Published: January 2020

Methods were developed for the determination of oligoalanine and other short-chain peptides and peptide analogs in ruminal fluid by using reverse-phase high-pressure liquid chromatography. Chromatographic analysis of the breakdown of (Ala)3 and (Ala)4 in ruminal fluid in vitro revealed that the predominant mechanism of hydrolysis was a dipeptidyl peptidase-like activity. Hydrolysis proceeded from the N terminal of the peptide chain; N-acetyl-(Ala)3 was broken down at 11% of the rate of breakdown of (Ala)3 or (Ala)3-p-nitroanilide. (Ala)2-p-nitroanilide was hydrolyzed most rapidly of the arylamide substrates tested, but fluorogenic 4-methoxy-2-naphthylamide (MNA) compounds were more convenient and potentially more versatile substrates than p-nitroanilides. Gly-Arg-MNA was the most rapidly hydrolyzed dipeptidyl peptidase substrate, suggesting that ruminal peptidase activity was predominantly of a type I specificity.
DNA-DNA homology among lactose- and sucrose-fermenting transconjugants from Lactococcus lactis strains exhibiting reduced bacteriophage sensitivity
Authors: Steele J L , Murphy M C , Daly C , McKay L L
Published: January 2020

DNA-DNA homology between a reduced bacteriophage sensitivity (Rbs+) probe and DNA from both Rbs+ and Rbs- Lactococcus lactis strains was examined. Homology was detected between the probe and five plasmids (pCI750, pCC34, pEB56, pNP2, and pJS88) isolated from lactose-positive Rbs+ transconjugants and between the probe and genomic DNA of a sucrose-positive Rbs+ transconjugant. Additionally, hybridizations conducted between the probe and plasmids reported to encode abortive bacteriophage infection indicated homology with pTR2030 but not with pBF61 and pGBK17. The results suggest that a common genetic determinant(s) may be present in a variety of lactococcal plasmids coding for Rbs+.
The conjugative plasmid pTR2030 encodes two bacteriophage defense mechanisms in lactococci, restriction modification (R+/M+) and abortive infection (Hsp+)
Authors: Hill C , Pierce K , Klaenhammer T R
Published: January 2020

pTR2030 is a conjugative plasmid which encodes resistance to bacteriophage in lactococci by a mechanism that aborts the phage infection (Hsp+). Subcloning and in vivo deletion events showed that two independent mechanisms of resistance are located on a 13.6-kilobase Bg/II fragment cloned in pSA3; one mechanism is responsible for the abortive infection, and the other incodes a restriction modification system. The introduction of pTR2030 or the recombinant plasmid pTK6 resulted in the loss of a resident restriction modification plasmid in Lactococcus lactis NCK202 which was not previously identified.
Microbiological Comparison of Surface Soil and Unsaturated Subsurface Soil from a Semiarid High Desert
Authors: Colwell Frederick S.
Published: January 2020

Thirty-two chemoheterotrophic bacteria were isolated from unsaturated subsurface soil samples obtained from ca. 70 m below land surface in a high desert in southeastern Idaho. Most isolates were gram positive (84%) and strict aerobes (79%). Acridine orange direct counts of microbes in one subsurface sample showed lower numbers than similar counts performed on surface soils from the same location (ca. 5 × 10
5
versus 2 × 10
6
cells per g [dry weight] of soil), but higher numbers than those from plate counts performed on the subsurface material. Another sample taken from the same depth at another location showed no evidence of colonies under identical conditions. Soil analyses indicated that subsurface sediments versus surface soils were slightly alkaline (pH 7.9 versus 7.4), had a higher water content (25.7 versus 6.3%), and had lower organic carbon concentrations (0.05 to 0.17 versus 0.25% of soil dry weight). Analyses of biologically relevant gases from the unsaturated subsurface indicated an aerobic environment. As in other unsaturated soil environments, either a high proportion of bacteria in these subsurface sediments are not viable or they are incapable of growth on conventional media under aerobic conditions. The presence and numbers of bacteria in these deep sediments may be influenced by colonization opportunities afforded by periodic percolation of surface water through fractures in overlying strata.
Development of an Adhesion Assay and Characterization of an Adhesion-Deficient Mutant of
Pseudomonas fluorescens
Authors: DeFlaun Mary F. , Tanzer Adam S. , McAteer Allison L. , Marshall Bonnie , Levy Stuart B.
Published: January 2020

A sand column adhesion assay was developed which distinguishes the adhesion abilities of a number of pseudomonads isolated from fine sandy loam.
Pseudomonas fluorescens
Pf0-1 which adhered at >90% of the total cells added was subjected to transposon Tn
5
insertion mutagenesis. From 2,500 Pf0-1::Tn
5
mutants examined in the sand column assay, two adhesion-deficient Pf0-1 mutants showing <50% attachment were isolated. Marker exchange analysis of one of these mutants, Pf0-5, confirmed that the decreased adhesion was linked to the Tn
5
insertion in the chromosome. The growth rate of Pf0-5 in enriched media and sterile soil was similar to that of the wild type; in minimal medium, however, Pf0-5 grew faster. In a soil column assay, less Pf0-5 than wild-type bacteria were recovered, suggesting a decreased ability to persist in soil. A 34-kilodalton major outer membrane protein present in the wild type was missing in Pf0-5. Transmission electron microscopy of the cell surface revealed that the wild-type possessed polar flagella which were absent in the mutant.
Sulfur-Containing Amino Acids as Precursors of Thiols in Anoxic Coastal Sediments
Authors: Kiene Ronald P. , Malloy Kirk D. , Taylor Barrie F.
Published: January 2020

Sulfur-containing amino acids were examined as precursors for thiols in anoxic coastal sediments. Substrates (10 to 100 μM) were anaerobically incubated with sediment slurries; thiols were assayed as isoindole derivatives by high-performance liquid chromatography; and microbial transformations of thiols, in contrast to their chemical binding by sediment particles, were identified by inhibition with a mixture of chloramphenicol and tetracycline. Methionine and homocysteine were transformed to methanethiol and 3-mercaptopropionate (3-MPA); methionine stimulated mainly methanethiol production, whereas homocysteine generated more 3-MPA than methanethiol. 2-Keto-4-methiolbutyrate yielded results similar to those with methionine, indicating that demethiolation yields methanethiol at the keto-acid level. Glutathione gave rise to cysteine, which was further transformed to 3-mercaptopyruvate and thence to mercaptoacetate and mercaptoethanol. Mercaptoethanol was oxidized to mercaptoacetate, which was biologically consumed. In conclusion, sulfurcontaining amino acids contribute to the range of thiols that occur in anoxic coastal sediments. New metabolic and environmental transformations were identified: the production of 3-MPA as a metabolite of methionine and the transformation of mercaptopyruvate to mercaptoethanol and mercaptoacetate.
Identification of the Minimal Replicon of
Lactococcus lactis
subsp.
lactis
UC317 Plasmid pCI305
Authors: Hayes Finbarr , Daly Charles , Fitzgerald Gerald F.
Published: January 2020

Replication functions of the stable, cryptic 8.7-kilobase (kb) plasmid pCI305 from multi-plasmid-containing
Lactococcus lactis
subsp.
lactis
UC317 were studied. Analysis of this replicon was facilitated by the construction of replication probe vectors that consisted of the pBR322 replication region, a pUC18-derived multiple cloning site, and either the
cat
gene of pC194 (pCI341; 3.1 kb) or the
erm
gene of pAMβ1 (pCI3330; 4.0 kb). Plasmid pCI305 was introduced into plasmid-free
L. lactis
subsp.
lactis
MG1363Sm, a streptomycin-resistant derivative of MG1363, by a transformation procedure with the 75-kb lactose-proteinase plasmid pCI301 of UC317 as a marker plasmid. A combination of transposon Tn
5
mutagenesis and subcloning in pCI341 and pCI3330 with individual Tn
5
insertions around the replication region facilitated the identification of a 1.6-kb minimal replicon on pCI305. This region was separable into two domains: (i) a 1.3-kb region (
repB
) encoding a
trans
-acting function (in vitro transcription-translation studies suggested the involvement of a 48-kilodalton protein); and (ii) a 0.3-kb region (
repA
) sufficient to direct replication when provided with
repB
in
trans
and thus probably containing the origin of replication. Lactococcus-
Escherichia coli
shuttle vectors based on the pCI305 replication region were constructed.
Exoprotease Activity of Two Marine Bacteria during Starvation
Authors: Albertson Nan H. , Nyström Thomas , Kjelleberg Staffan
Published: January 2020

Exoprotease activity during 120 h of total energy and nutrient starvation was examined in two marine bacteria,
Vibrio
sp. strain S14 and
Pseudomonas
sp. strain S9. The activity was determined by spectrophotometric measurement of the rate of release of soluble color from an insoluble azure dye derivative of hide powder (hide powder azure). Starved cells of both strains (5 h for S14, and 4 or 24 h for S9) showed greater extracellular proteolytic activity than at the onset of starvation. The exoprotease activity of cells starved for longer periods of time then decreased, but was found to be present at significant levels throughout the starvation period studied (120 h). The accumulation of exoprotease activity in the bulk phase during starvation indicated that both strains constitutively excreted extracellular proteases. As deduced from experiments with chloramphenicol, de novo protein synthesis during starvation was required for the production and/or release of the exoproteases into the surrounding environment. The degradation of hide powder azure allowed an immediate increase in respiration rate, also by long-term-starved cells. This suggests that metabolic systems are primed to respond to the availability of substrates, allowing the cells to recover rapidly. The regulation of exoprotease activity was also studied and found to be different in the two strains. Casamino Acids repressed exoprotease activity in
Pseudomonas
sp. strain S9, whereas a mechanism similar to catabolite repression was found for
Vibrio
sp. strain S14 in that glucose repressed activity and cyclic AMP reversed this effect. The exoproteases appeared to be metalloproteinases because the addition of EDTA to cell-free starvation supernatants from both strains significantly inhibited the activity of the proteases.
Efficiencies of Recovery of Bdellovibrios from Brackish- Water Environments by Using Various Bacterial Species as Prey
Authors: Schoeffield A. J. , Williams H. N.
Published: January 2020

A total of 44 bacterial species subdivided into 10 trial experiments have been used as prey for the recovery of bdellovibrios from samples of water from a brackish tidal pond and an aquarium saltwater tank. In an initial investigation, the recovery efficiency of each of the test bacterial species was compared with that of a designated standard prey,
Vibrio parahaemolyticus
P-5. The results revealed that in each case strain P-5 yielded an equal or significantly greater number of plaques of bdellovibrios than the test prey with but a single exception, strain CS5. In repeat experiments, CS5 yielded fewer plaques than P-5. To determine whether the use of multiple bacterial species compared with a single species as prey would increase the number of PFU of bdellovibrios recovered, material from plaques appearing on each of the test prey in the respective trials was sequentially subcultured onto two respective agar plates, the first containing as prey
V. parahaemolyticus
P-5 and the second containing the initial test organism. In nearly every case, subculture of plaques from lawns of the test prey to P-5 resulted in plaque formation. On the basis of the results, the use of several test prey and P-5 did not result in the recovery of any more bdellovibrio PFU than the use of P-5 alone. In this study,
V. parahaemolyticus
P-5 was observed to be the most efficient prey for the recovery of bdellovibrios from moderate salt water.
Effects of Acid Stress on Aerobic Decomposition of Algal and Aquatic Macrophyte Detritus: Direct Comparison in a Radiocarbon Assay
Authors: Schoenberg Steven A. , Benner Ronald , Armstrong Anthony , Sobecky Patricia , Hodson Robert E.
Published: January 2020

Radiolabeled phytoplankton and macrophyte lignocelluloses were incubated at pHs 4 and 7 in water from a naturally acidic freshwater wetland (Okefenokee Swamp; ambient pH, 3.8 to 4.2), a freshwater reservoir (L-Lake; pH 6.7 to 7.2), and a marine marsh (Sapelo Island; pH ∼7.8). The data suggest that acidity is an important factor in explaining the lower decomposition rates of algae in Okefenokee Swamp water relative to L-Lake or Sapelo Island water. The decomposition of algal substrate was less sensitive to low pH (∼5 to 35% inhibition) than was the decomposition of lignocellulose (∼30 to 70% inhibition). These substrate-dependent differences were greater and more consistent in salt marsh than in L-lake incubations. In both freshwater sites, the extent to which decomposition was suppressed by acidity was greater for green algal substrate than for mixed diatom or blue-green algal (cyanobacteria) substrates. The use of different bases to adjust pH or incubation in a defined saltwater medium had no significant effect on substrate-dependent differences. Although pH differences with lignocellulose were larger in marine incubations, amendment of lakewater with marine bacteria or with calcium, known to stabilize exoenzymes in soils, did not magnify the sensitivity of decomposition to acid stress.
Mannitol Metabolism in
Lentinus edodes
, the Shiitake Mushroom
Authors: Kulkarni Rajiv K.
Published: January 2020

Mannitol metabolism was evaluated in fruiting bodies of
Lentinus edodes.
Cell extracts were prepared from fruiting bodies, and key enzymes involved in mannitol metabolism were assayed, including hexokinase, mannitol dehydrogenase, mannitol-1-phosphate dehydrogenase, mannitol-1-phosphatase, and fructose-6-phosphatase. Mannitol dehydrogenase, fructose-6-phosphatase, mannitol-1-phosphatase, and hexokinase activities were found in extracts of fruiting bodies. However, mannitol-1-phosphate dehydrogenase activity was not detected. Mycelial cultures were grown in an enriched liquid medium, and enzymes of the mannitol cycle were assayed in cell extracts of rapidly growing cells. Mannitol-1-phosphate dehydrogenase activity was also not found in mycelial extracts. Hence, evidence for a complete mannitol cycle both in vegetative mycelia and during mushroom development was lacking. The pathway of mannitol synthesis in
L. edodes
appears to utilize fructose as an intermediate.
Initial Test of the Benchmark Chemical Approach for Predicting Microbial Transformation Rates in Aquatic Environments
Authors: Newton Thomas D. , Gattie David K. , Lewis David L.
Published: January 2020

Using 2,4-dichlorophenoxyacetic acid methyl ester (2,4-DME) as a benchmark chemical, we determined relative pseudo-first-order rate coefficients for the butoxyethyl ester of 2,4-dichlorophenoxyacetic acid (2,4-DBE), methyl parathion, and methyl-3-chlorobenzoate in a diversity of microbial samples, including water, sediment, biofilm, and floating microbial mats collected from a laboratory mesocosm as well as from streams, lakes, and wetlands in Georgia and Florida. The decreasing order of reactivity for relative microbial transformation rates was 2,4-DBE > 2,4-DME > methyl-3-chlorobenzoate > methyl parathion. Half-lives of the chemicals varied about 60-fold depending on the chemical and microbial sample. Relative rate coefficients, however, typically varied only about threefold for field-collected samples. Relative rate coefficients determined with samples from a laboratory mesocosm were consistently low compared with the field sample data. Overall, the data indicated that microbial transformation rates of a chemical can be satisfactorily inferred for a wide variety of microbial habitats—such as water, biofilm, or a sediment—on the basis of its transformation rate relative to that of an appropriate benchmark chemical by using a single type of microbial sample.
Carbon Flow in Mercury Biomethylation by
Desulfovibrio desulfuricans
Authors: Berman Mitchell , Chase Theodore , Bartha Richard
Published: January 2020

Radiocarbon incorporation from pyruvate and serine into monomethylmercury by
Desulfovibrio desulfuricans
was consistent with the proposal that the methyl group originates from C-3 of serine. Immunodiagnostic assays measured 4 to 35 μg of tetrahydrofolate and 58 to 161 ng of cobalamin or a closely related cobalt porphyrin per g of cell protein in
D. desulfuricans.
The light-reversible inhibition of mercury methylation by propyl iodide in
D. desulfuricans
indicates methyl transfer by a cobalt porphyrin.
New plate medium for facilitated differentiation of Salmonella spp. from Proteus spp. and other enteric bacteria
Authors: Rambach A
Published: January 2020

A new agar medium for the differentiation of Salmonella spp. from other members of the family Enterobacteriaceae is described. This medium exploits a novel phenotypic characteristic of Salmonella spp.: the formation of acid from propylene glycol. This characteristic may be used in combination with a chromogenic indicator of beta-galactosidase to differentiate Salmonella spp. from Proteus spp. and the other members of the Enterobacteriaceae. Desoxycholate may be included in the plate medium as an inhibitor of gram-positive organisms. Non-typhi Salmonella spp. yield distinct, bright red colonies on this medium, allowing facilitated identification and unambiguous differentiation from Proteus spp.
Organomercurial-volatilizing bacteria in the mercury-polluted sediment of Minamata Bay, Japan
Authors: Nakamura K , Sakamoto M , Uchiyama H , Yagi O
Published: January 2020

A total of 4,604 bacterial strains isolated from the sediments of Minamata Bay and nearby low-level-mercury stations (control stations) were screened for the ability to volatilize mercury from inorganic and organic mercurial compounds. The strains that volatilize mercury from several kinds of organomercurials were found only in the sediments of Minamata Bay.
Microbial colonization of aquifer sediment exposed in a groundwater well in northern Germany
Authors: Hirsch P , Rades-Rohkohl E
Published: January 2020

Microbial growth within the water-saturated subsurface environment was investigated by exposing sandy sediments to groundwater for 12 weeks at a depth of 10 or 20 m in a stainless-steel groundwater well. Washing and heating the sediment to 600 degrees C (removal of organic carbon) prior to the exposure did not prevent the natural microbial community from colonizing the sterilized sediment samples. Total cell counts of more than 10(7) or 10(8) per g of dried sediment were obtained. Viable cell counts of 10(5) cells per g on oligotrophic media indicated the presence, within the exposed sediment, of a highly active and multiplying biota. Microscopic analysis of enrichments inoculated with exposed sediment samples revealed a total of 45 different morphotypes, approximately 42% of the microbial community observed in previous studies of this site. The interstitial water running off of the retrieved sediment contained only 17 morphotypes and had up to 6 x 10(5) viable cells per ml.
Fusarochromanone production by Fusarium isolates
Authors: Wu W D , Nelson P E , Cook M E , Smalley E B
Published: January 2020

Sixty two Fusarium isolates representing nine species from many parts of the world were screened for fusarochromanone production. A simplified method for the detection of fusarochromanone in culture filtrates or grain cultures was used. Under UV irradiation (364 nm) the chloroform phase from fusarochromanone-positive culture extracts fluoresced a characteristic bright blue color. Results were confirmed by thin-layer-chromatography comparison with pure fusarochromanone standards. Detection was possible in cultures as young as 1 week old. Biosynthesis of fusarochromanone was rare in Fusarium spp. and was only detected in three isolates of Fusarium equiseti, namely R-4482 (barley [Federal Republic of Germany]), R-6137 (barley [Alaska]), and R-8508 (potato [Denmark]), among all the isolates tested from various geographic sources.
Extracellular proteases from Xanthomonas campestris pv. campestris, the black rot pathogen
Authors: Dow J M , Clarke B R , Milligan D E , Tang J L , Daniels M J
Published: January 2020

Two proteases (PRT1 and PRT2) were fractionated from culture supernatants of wild-type Xanthomonas campestris pv. campestris by cation-exchange chromatography on SP-5PW. Inhibitor experiments showed that PRT 1 was a serine protease which required calcium ions for activity or stability or both and that PRT 2 was a zinc-requiring metalloprotease. PRT 1 and PRT 2 showed different patterns of degradation of beta-casein. The two proteases comprised almost all of the extracellular proteolytic activity of the wild type. A protease-deficient mutant which lacked both PRT 1 and PRT 2 showed considerable loss of virulence in pathogenicity tests when bacteria were introduced into mature turnip leaves through cut vein endings. This suggests that PRT 1 and PRT 2 have a role in black rot pathogenesis.
Occurrence of Gibberella zeae strains that produce both nivalenol and deoxynivalenol
Authors: Sugiura Y , Watanabe Y , Tanaka T , Yamamoto S , Ueno Y
Published: January 2020

By single ascospore isolation, several sets of asci containing eight ascospores were isolated from perithecia of Gibberella zeae. Of these sets, seven were investigated for their ability to produce 8-ketotrichothecene mycotoxins on rice grains. Analyses were made with gas chromatography-mass spectrometry and gas chromatography with 63Ni electron capture detection. Of 56 total isolates, 11 produced nivalenol, 4-acetylnivalenol, and deoxynivalenol, 1 produced nivalenol and deoxynivalenol, 7 produced deoxynivalenol and 3-acetyldeoxynivalenol, 19 produced deoxynivalenol and 15-acetyldeoxynivalenol, and 6 produced deoxynivalenol and both 15- and 3-acetyldeoxynivalenol. The remaining 12 isolates produced nivalenol and 4-acetylnivalenol. All isolates of G. zeae that we examined could produce 8-ketotrichothecenes in this investigation. This report is the first to demonstrate the presence of G. zeae isolates producing both nivalenol and deoxynivalenol. In addition, differences in the production between 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol are discussed in relation to culture conditions.
Pulsed-Field Gel Electrophoresis of
Sma
I Digests of Lactococcal Genomic DNA, a Novel Method of Strain Identification
Authors: Tanskanen Eva I. , Tulloch Debra L. , Hillier Alan J. , Davidson Barrie E.
Published: January 2020

The pulsed-field gel electrophoresis (PFGE) pattern of
Sma
I digests of 29 strains of
Lactococcus lactis
subsp.
lactis
and subsp.
cremoris
were determined. Unrelated strains yielded markedly different patterns of digestion products. Bacteriophage-resistant derivatives of four strains, generated by a method analogous to that used regularly in some cheese factories, yielded patterns that were identical or almost identical to that of the parent strain. It is proposed that a 16-h PFGE run with a pulse time increasing linearly from 1 to 20 s, which separates fragments between 50 and 240 kilobase pairs (kbp) and produces a pattern containing around 15 bands, can be used as a reliable procedure for strain identification in the lactococci.
Sma
I digests of 24 of the strains were analyzed by PFGE at three different pulse times to determine accurately the sizes of fragments bigger than 8 kbp. The sum of the sizes of all of the fragments in the digest of a strain provided an estimate of the genome size of the strain. For all the strains analyzed, this estimate was within the range of 2.0 to 2.7 Mbp, with no apparent difference between
L. lactis
subsp.
lactis, L. lactis
subsp.
lactis
biovar diacetylactis and
L. lactis
subsp.
cremoris
strains.
celB, a gene coding for a bifunctional cellulase from the extreme thermophile "Caldocellum saccharolyticum"
Authors: Saul D J , Williams L C , Grayling R A , Chamley L W , Love D R , Bergquist P L
Published: January 2020

"Caldocellum saccharolyticum" is an obligatory anaerobic thermophilic bacterium. A gene from this organism, designated celB, has been cloned in Escherichia coli as part of a bacteriophage lambda gene library. This gene produces a thermostable cellulase that shows both endoglucanase and exoglucanase activities on test substrates and is able to degrade crystalline cellulose to glucose. The sequence of celB has homology with both exo- and endoglucanases described by others. It appears to have a central domain without enzymatic activity which is joined to the enzymatic domains by runs of amino acids rich in proline and threonine (PT boxes). Deletion analysis shows that the exoglucanase activity is located in the amino-terminal domain of the enzyme and that endoglucanase activity is located in the carboxy-terminal domain. There are internal transcriptional and translational start sites within the gene. The intact gene has been cloned into a temperature-inducible expression vector, pJLA602, and overexpressed in E. coli. Polyacrylamide gel electrophoresis showed that celB produced a protein with a molecular weight of 118,000 to 120,000. A number of smaller proteins with activity against carboxymethyl cellulose and 4-methyl umbelliferyl-beta-D-cellobioside were also produced. These are believed to be the result of alternative translational start sites and/or proteolytic degradation products of the translated gene product.
Biosynthesis of Poly-β-Hydroxyalkanoates from Pentoses by
Pseudomonas pseudoflava
Authors: Bertrand J.-L. , Ramsay B. A. , Ramsay J. A. , Chavarie C.
Published: January 2020

The potential of
Pseudomonas pseudoflava
to produce poly-β-hydroxyalkanoates (PHAs) from pentoses was studied. This organism was able to use a hydrolysate from the hemicellulosic fraction of poplar wood as a carbon and energy source for its growth. However, in batch cultures, growth was inhibited completely at hydrolysate concentrations higher than 30% (vol/vol). When
P. pseudoflava
was grown on the major sugars present in hemicelluloses in batch cultures, poly-β-hydroxybutyric acid (PHB) accumulated when glucose, xylose, or arabinose was the sole carbon source, with the final PHB content varying from 17% (wt/wt) of the biomass dry weight on arabinose to 22% (wt/wt) of the biomass dry weight on glucose and xylose. Specific growth rates were 0.58 h
−1
on glucose, 0.13 h
−1
on xylose, and 0.10 h
−1
on arabinose, while the specific PHB production rates based on total biomass ranged from 0.02 g g
−1
h
−1
on arabinose to 0.11 g g
−1
h
−1
on glucose. PHB weight-average molecular weights were 640,000 on arabinose and 1,100,000 on glucose and xylose. The absolute amount of PHB in the cells decreased markedly when nitrogen limitation was relaxed by feeding ammonium sulfate at the end of the PHB accumulation stage of the arabinose and xylose fermentations. Copolymers of β-hydroxybutyric and β-hydroxyvaleric acids were produced when propionic acid was added to shake flasks containing 10 g of glucose liter
−1
. The β-hydroxyvaleric acid monomer content attained a maximum of 45 mol% when the initial propionic acid concentration was 2 g liter
−1
.
Dynamics of ruminal ciliated protozoa in feedlot cattle
Authors: Towne G , Nagaraja T G , Brandt R T , Kemp K E
Published: January 2020

Fluctuations in ciliated protozoan concentrations were monitored in 40 individually fed crossbred heifers that were stepped up to an 85% concentrate diet either slowly (12 days) or rapidly (3 days), with or without monensin (30 ppm). Ruminal fluid was withdrawn from all animals by stomach tube at the start of the study, after each group reached full feed, and at 14-day intervals thereafter throughout the finishing period until termination (day 119). Neither monensin nor speed of step-up affected (P greater than 0.10) total protozoan concentrations, ruminal pH, or lactic acid concentrations. Average protozoan concentrations peaked on day 5, progressively declined until day 56, and then increased (P less than 0.05), suggesting an adaptation to ruminal conditions. Concentrations of Isotricha spp. were higher (P less than 0.05) on the final two sampling dates than at any other time. After day 28, Entodinium, Isotricha, and Polyplastron were the only surviving genera. Protozoa were not detected in 11 heifers on day 42 and day 56, but only two animals were defaunated on day 119, indicating either exogenous or endogenous refaunation. Average protozoan concentrations were not different (P greater than 0.25) between ruminal samples collected by stomach tube the day before slaughter (2.8 x 10(5)/g) and digesta samples collected the next day (1.6 x 10(5)/g). In feedlot cattle, defaunation apparently is transitory and individual animals harbor a dynamic protozoan population that fluctuates in response to changing ruminal conditions.
Thiophene-degrading Escherichia coli mutants possess sulfone oxidase activity and show altered resistance to sulfur-containing antibiotics
Authors: Juhl M J , Clark D P
Published: January 2020

We have previously isolated mutants of Escherichia coli which show increased oxidation of heterocyclic furan and thiophene substrates. We have now found that strains carrying the thdA mutation express a novel enzyme activity which oxidizes a variety of substrates containing a sulfone (SO2) moiety. Both heterocyclic sulfones (e.g., tetramethylene sulfone) and simple aliphatic sulfones (e.g., ethyl sulfone) were oxidized. The thdA mutants were more resistant than wild-type strains to aromatic sulfone antibiotics such as dapsone. In contrast they showed increased susceptibility to thiolutin, a cyclic antibiotic containing sulfur at the sulfide level of oxidation. Several new thdA mutant alleles were isolated by selecting for increased oxidation of various aliphatic sulfur compounds. These new thdA mutants showed similar sulfone oxidase activity and the same map location (at 10.7 min) as the original thdA1 mutation. The constitutive fadR mutation was required for the phenotypic expression of thdA-mediated oxidation of sulfur compounds. However, the thdA-directed expression of sulfone oxidase activity was not fadR dependent. The thdC and thdD mutations probably protect against the toxicity of thiophene derivatives rather than conferring improved metabolic capability.