bluuebunny/crossref_metadata_2025 huggingface dataset:
Formation of Methane and Carbon Dioxide from Dimethylselenide in Anoxic Sediments and by a Methanogenic Bacterium
Authors: Oremland Ronald S. , Zehr Jon P.
Published: January 2020

Anaerobic San Francisco Bay salt marsh sediments rapidly metabolized [
14
C]dimethylselenide (DMSe) to
14
CH
4
and
14
CO
2
. Addition of selective inhibitors (2-bromoethanesulfonic acid or molybdate) to these sediments indicated that both methanogenic and sulfate-respiring bacteria could degrade DMSe to gaseous products. However, sediments taken from the selenium-contaminated Kesterson Wildlife Refuge produced only
14
CO
2
from [
14
C]DMSe, implying that methanogens were not important in the Kesterson samples. A pure culture of a dimethylsulfide (DMS)-grown methylotrophic methanogen converted [
14
C]DMSe to
14
CH
4
and
14
CO
2
. However, the organism could not grow on DMSe. Addition of DMS to either sediments or the pure culture retarded the metabolism of DMSe. This effect appeared to be caused by competitive inhibition, thereby indicating a common enzyme system for DMS and DMSe metabolism. DMSe appears to be degraded as part of the DMS pool present in anoxic environments. These results suggest that methylotrophic methanogens may demethylate methylated forms of other metals and metalloids found in nature.
Anaerobic oxidation of p-cresol by a denitrifying bacterium
Authors: Bossert I D , Young L Y
Published: January 2020

Metabolism of p-cresol (pCr) under nitrate-reducing conditions is mediated by the denitrifying bacterial isolate PC-07. The methyl substituent of the substrate is oxidized anaerobically by whole-cell suspensions of PC-07 through a series of dehydrogenation and hydration reactions to yield p-hydroxybenzoate (pOHB) in stoichiometric proportions. The partially oxidized intermediates in the pathway p-hydroxybenzyl alcohol and p-hydroxybenzaldehyde can also serve as substrates for pOHB formation. Nitrate is required as the external electron acceptor and is reduced to molecular N2. Reduction of the nitrate is stoichiometric, with pCr serving as the electron donor. In addition, the molar relationship between the electron acceptor (NO3-) reduced to the electron donor oxidized decreased to approximately 2:3 and then to 1:3 when p-hydroxybenzyl alcohol or p-hydroxybenzaldehyde, respectively, served as substrates. The decreased ratios were to be expected when the partially oxidized intermediates served as substrates, because they provided correspondingly less reducing power for pOHB formation. The anaerobic oxidation of pCr by PC-07 demonstrates a mechanism whereby aromatic compounds can be transformed in anoxic environments.
Evaluation of Diverse Antisera, Conjugates, and Support Media for Detecting
Bradyrhizobium japonicum
by Indirect Enzyme-Linked Immunosorbent Assay
Authors: Ayanaba Abateni , Weiland Karen D. , Zablotowicz Robert M.
Published: January 2020

We evaluated three antisera and four enzyme conjugates for the detection of
Bradyrhizobium japonicum
by an indirect enzyme-linked immunosorbent assay in microtiter plates. Nitrocellulose membrane sheets were then evaluated as an alternative support medium by using some combinations. Partially purified immunoglobulin G (IgG) or unpurified antisera to strain USDA 110 raised in rabbits, goats, or sheep was reacted in microtiter plates with alkaline phosphatase conjugated to protein A, goat anti-rabbit (GAR), sheep anti-rabbit (SAR), or rabbit anti-goat (RAG) IgG. Cultures or nodules containing homologous rhizobia were detected with equal sensitivity when protein A, GAR, or SAR was reacted with 5 μg of protein IgG per ml or a 1:800 titer of antisera from rabbits, but not goats or sheep. RAG reacted with IgG or antisera from goats or sheep. The detection limit was 2 × 10
5
rhizobia per well. Rhizobia were spotted on nitrocellulose sheets as an alternative support medium, followed by soaking in 5 μg of protein per ml as IgG and 1:4,000 dilutions of protein A or GAR conjugate. Rhizobia in serogroup 110 were detected with the dye combination Nitro Blue Tetrazolium-5-bromo-4-chloro-3-indolyl phosphate (NBT-BCIP), and rhizobia in serogroup 122 were detected with fast red-naphthol phosphate (FR-NP). At the conclusion of the 5-h assay, purple (NBT-BCIP) or red (FR-NP) spots were visible in positive reactions. The sensitivity of detection was about 1,000 rhizobial cells or 3 μg of nodules tissue.
Structure and Dynamics of Anaerobic Bacterial Aggregates in a Gas-Lift Reactor
Authors: Beeftink H. H. , Staugaard P.
Published: January 2020

Anaerobic mixed-culture aggregates, which converted glucose to acetic, propionic, butyric, and valeric acids, were formed under controlled conditions of substrate feed (carbon limitation) and hydraulic regimen. The continuous-flow system used (anaerobic gas-lift reactor) was designed to retain bacterial aggregates in a well-mixed reactor. Carrier availability (i.e., liquid-suspended sand grains) proved necessary for bacterial aggregate formation from individual cells during reactor start-up. Electron microscopic examination revealed that incipient colonization of sand grains by bacteria from the bulk liquid occurred in surface irregularities, conceivably reflecting local quiescence. Subsequent confluent biofilm formation on sand grains proved to be unstable, however. Substrate depletion in the bulk liquid is assumed to weaken deeper parts of the biofilm due to cellular lysis, after which production of gas bubbles and liquid shearing forces cause sloughing. The resulting fragments, although sand free, were nevertheless large enough to be retained in the reactor and gradually grew larger through bacterial growth and by clumping together with other fragments. In the final steady state, high cell densities were maintained in the form of aggregates, while sand had virtually disappeared due to sampling losses and wash-out. Numerical cell densities within aggregates ranged from 10
12
/ml at the periphery to very low values in the center. The cells were enmeshed in a polymer matrix containing polysaccharides; nevertheless, carbon sufficiency was not a prerequisite to sustain high hold-up ratios.
Steroid desmolase synthesis by Eubacterium desmolans and Clostridium cadavaris
Authors: Bokkenheuser V D , Winter J , Morris G N , Locascio S
Published: January 2020

The synthesis of a steroid desmolase was demonstrated in two obligate anaerobes: a new bacterial species, Eubacterium desmolans, isolated from cat fecal flora, and Clostridium cadavaris, recovered from sewage of New York City. The enzyme cleaves the C-17-C-20 bond of corticoids possessing hydroxyl functions at C-17 and C-21. The conversion is quantitative, provided the substrate concentration is less than 100 micrograms/ml and the organisms are in the log phase. The velocity of transformation parallels the bacterial growth curve and in the log phase is higher for E. desmolans than for C. cadavaris. In addition, both organisms synthesize a 20 beta-hydroxysteroid dehydrogenase.
Localization of inclusion bodies in Escherichia coli overproducing beta-lactamase or alkaline phosphatase
Authors: Georgiou G , Telford J N , Shuler M L , Wilson D B
Published: January 2020

High-level synthesis of the periplasmic protein beta-lactamase in Escherichia coli caused the formation of insoluble protein precipitates called inclusion bodies. beta-Lactamase inclusion bodies differed from those reported previously in that they appeared to be localized in the periplasmic space, not in the cytoplasm. The inclusion bodies contained mature beta-lactamase and were solubilized more easily than has been reported for cytoplasmic inclusion bodies. In contrast, overproduction of the periplasmic protein alkaline phosphatase caused the formation of cytoplasmic inclusion bodies containing alkaline phosphatase precursor.
Growth Characteristics of a Novel Nitrogen-Fixing Cellulolytic Bacterium
Authors: Greene Richard V. , Freer Shelby N.
Published: January 2020

Growth characteristics of a cellulolytic nitrogen-fixing bacterium isolated from a marine shipworm by Waterbury et al. (J. B. Waterbury, C. B. Calloway, and R. D. Turner, Science 221:1401-1403, 1983) are described. When grown microaerobically, the bacterium exhibited doubling times of about 2 days in cellulose-supplemented synthetic medium devoid of combined nitrogen. Maximum growth was reached 12 to 16 days after inoculation. Growth optima for pH, temperature, and NaCl concentration were 8.5, 30 to 35°C, and 0.3 M, respectively. During growth the bacterium produced succinic acid (0.026%) and acetic acid (0.010%). Formic acid (0.010%) was produced during the stationary growth phase. No growth was observed when glucose was the sole carbon source. Cellobiose supported weak growth, while longer-chain-length cellodextrins supported extensive growth. Analysis of residual carbohydrates in the medium during growth indicated that the bacterium catabolized a terminal glucose moiety from the cellodextrin chain.
Denitrification in Marl and Peat Sediments in the Florida Everglades
Authors: Gordon A. S. , Cooper W. J. , Scheidt D. J.
Published: January 2020

The potential for denitrification in marl and peat sediments in the Shark River Slough in the Everglades National Park was determined by the acetylene blockage assay. The influence of nitrate concentration on denitrification rate and N
2
O yield from added nitrate was examined. The effects of added glucose and phosphate and of temperature on the denitrification potential were determined. The sediments readily denitrified added nitrate. N
2
O was released from the sediments both with and without added acetylene. The marl sediments had higher rates than the peat on every date sampled. Denitrification was nitrate limited; however, the yields of N
2
O amounted to only 10 to 34% of the added nitrate when 100 μM nitrate was added. On the basis of measured increases in ammonium concentration, it appears that the balance of added nitrate may be converted to ammonium in the marl sediment. The sediment temperature at the time of sampling greatly influenced the denitrification potential (15-fold rate change) at the marl site, indicating that either the number or the specific activity of the denitrifiers changed in response to temperature fluctuations (9 to 25°C) in the sediment. It is apparent from this study that denitrification in Everglades sediments is not an effective means of removing excess nitrogen which may be introduced as nitrate into the ecosystem with supply water from the South Florida watershed and that sporadic addition of nitrate-rich water may lead to nitrous oxide release from these wetlands.
Inhibitor Evaluation with Immobilized
Nitrobacter agilis
Cells
Authors: Tsai Yu-Li , Schlasner Steven M. , Tuovinen Olli H.
Published: January 2020

Nitrobacter agilis
was entrapped in calcium alginate beads and used as a floating bed supplied with a continuous flow of nitrite medium. Complete nitrite oxidation was achieved within 30 h, and the system could be maintained for at least 210 h. The immobilized
Nitrobacter
system was subjected to sulfur oxyanions, acidity, and metal ions. Thiosulfate and tetrathionate (up to 20 mM each) did not inhibit the nitrite oxidation activity. A low pH of 4.2 resulted in the complete cessation of nitrite oxidation, and the activity was not restored upon increasing the pH to 7. Nitrite oxidation by
N. agilis
was sensitive to 10 mM each Ni
2+
and Al
3+
but insensitive to 10 mM MoO
4
2−
.
Mycological survey of Korean cereals and production of mycotoxins by Fusarium isolates
Authors: Lee U S , Jang H S , Tanaka T , Toyasaki N , Sugiura Y , Oh Y J , Cho C M , Ueno Y
Published: January 2020

The fungal species isolated from Korean cereals (barley, polished barley, wheat, rye, and malt) were Alternaria spp., Aspergillus spp., Chaetomium spp., Drechslera spp., Epicoccum sp., Fusarium spp., and Penicillium spp., etc. The number of Fusarium strains isolated was 36, and their ability to produce Fusarium mycotoxins on rice was tested. Nivalenol (NIV) was produced by Fusarium graminearum (7 of 9 isolates), Fusarium oxysporum (3 of 10 isolates), and Fusarium spp. (7 of 15 isolates). Of 15 isolates of Fusarium spp., 6 formed deoxynivalenol (DON). Fusarenon-X and 3-acetyl-DON were produced by most NIV- and DON-forming isolates, respectively. Zearalenone was produced by 3 isolates of F. graminearum, 1 isolate of Fusarium equiseti, and 11 isolates of Fusarium spp. T-2 toxin was not produced by any Fusarium isolates. The highest concentrations of mycotoxins produced by Fusarium isolates were 77.4 (NIV), 5.3 (DON), 138.3 (fusarenon-X), 40.6 (3-acetyl-DON), and 23.2 (zearalenone) micrograms/g.
Fixation, Counting, and Manipulation of Heterotrophic Nanoflagellates
Authors: Bloem Jaap , Bär-Gilissen Marie-José B. , Cappenberg Thomas E.
Published: January 2020

Quantitative effects of several fixatives on heterotrophic nanoflagellates (HNAN) and phototrophic nanoflagellates (PNAN) were investigated by hemacytometer and epifluorescence counting techniques. Counts of
Monas
sp. cultures before and after fixation with unbuffered 0.3% glutaraldehyde and 5% formaldehyde showed no loss of cells during fixation, and cell concentrations remained constant for several weeks after fixation. Buffering of fixatives with borax caused severe losses, up to 100% within 2 h. Field samples from Lake Vechten showed no decline of HNAN and total nanoflagellate concentrations for at least 1 week after fixation with 5% formaldehyde and with 1% glutaraldehyde. With 1% glutaraldehyde, the chlorophyll autofluorescence of PNAN was much brighter than with 5% formaldehyde, although it was lost after a few days and thus limited the storage time of samples. However, when primulin-stained slides were prepared soon after fixation and stored at −30°C, the loss of autofluorescence was prevented and PNAN and HNAN concentrations were stable for at least 16 weeks. Effects of filtration and centrifugation on HNAN were also studied. Filtration vacuum could not exceed 3 kPa since 10 kPa already caused losses of 15 to 20%. Similar losses were caused by centrifugation, even at low speed (500 ×
g
).
Environmental Parameters Regulating Gaseous Nitrogen Losses from Two Forested Ecosystems via Nitrification and Denitrification
Authors: Davidson Eric A. , Swank Wayne T.
Published: January 2020

Gaseous N losses from disturbed and reference forested watersheds at the Coweeta Hydrologic Laboratory in western North Carolina were studied by in situ N
2
O diffusion measurements and laboratory incubations throughout a 10-month period. Soil temperature, percent base saturation, and water-filled pore space accounted for 43% of the variation in in situ N
2
O diffusion measurements. Laboratory incubations distinguished the gaseous N products of nitrification and denitrification. Nitrifying activity, ambient NO
3
−
, and nitrification N
2
O were positively correlated with percent base saturation. However, differences between watersheds in soil N substrate caused by presence of leguminous black locust in the disturbed watershed were confounded with differences in soil acidity. Denitrification was most strongly affected by soil moisture, which in turn was determined by precipitation events and slope position. Gaseous N losses from well-drained midslope and toeslope landscape positions appeared to be minor relative to other N transformations. Favorable conditions for denitrification occurred at a poorly drained site near the stream of the disturbed watershed. Laboratory incubations revealed high rates of NO
3
−
reduction in these soils. We speculate that the riparian zone is a major site of depletion of NO
3
−
from the soil solution via denitrification.
Cadmium- and mercury-resistant Bacillus strains from a salt marsh and from Boston Harbor
Authors: Mahler I , Levinson H S , Wang Y , Halvorson H O
Published: January 2020

Bacteria resistant to cadmium or mercury or both were isolated from the Great Sippewissett Marsh (Cape Cod, Mass.) and from Boston Harbor. Many of these metal-resistant isolates were gram-positive aerobic sporeformers, although not necessarily isolated as spores. Although several of the isolated strains bore plasmids, cadmium and mercury resistances appeared to be, for the most part, chromosomally encoded. DNA sequence homology of the gram-positive cadmium- and mercury-resistant isolates was not demonstrable with metal resistance genes from plasmids of either gram-positive (pI258) or gram-negative (pDB7) origin. Cadmium resistance of all the marsh isolates tested resulted from reduced Cd2+ transport. On the other hand, three cadmium-resistant harbor isolates displayed considerable influx but no efflux of Cd2+. Hg-resistant strains detoxified mercury by transforming Hg2+ to volatile Hg0 via mercuric reductase.
Rapid serological identification of Vibrio vulnificus by anti-H coagglutination
Authors: Simonson J , Siebeling R J
Published: January 2020

Staphylococcus aureus Cowan 1 cells were armed with anti-flagellar (anti-H) antibody produced in rabbits immunized with flagellar core protein prepared from Vibrio vulnificus. This reagent was assessed by coagglutination for its capacity to agglutinate and identify V. vulnificus. A species-specific H antigen is expressed in the core proteins of the polar flagella of V. vulnificus. Of 435 V. vulnificus isolates identified bacteriologically, 432 (99.3%) were agglutinated in the slide test within 2 min after the addition of the anti-V. vulnificus H coagglutination reagent. Other than Vibrio pelagius, the reagent did not agglutinate 19 heterologous Vibrio spp. tested, including 290 V. cholerae, 22 V. mimicus, 395 V. parahaemolyticus, and 16 V. fluvialis isolates recovered from seafood and the marine environment. The serological resolution of the coagglutination reaction was enhanced if the organism under test was suspended in 0.1 M Tris buffer-0.1 mM EDTA-1.0% Triton X-100 (TET) for 24 h before serological examination. The TET buffer also increased the sensitivity of the coagglutination reaction 100-fold over that for isolates suspended in 0.3% formalinized phosphate-buffered saline before testing. The anti-H coagglutination test is a rapid, serologically specific, and inexpensive procedure for identifying V. vulnificus one step beyond primary isolation.
Isolation of Halotolerant
Thermus
spp. from Submarine Hot Springs in Iceland
Authors: Kristjansson Jakob K. , Hreggvidsson Gudmundur O. , Alfredsson Gudni A.
Published: January 2020

Thermophilic, aerobic bacteria of the genus
Thermus
were isolated from submarine alkaline hot springs in Iceland. Five submarine hot springs were sampled, and all had viable counts of
Thermus
spp. of about 10
3
CFU/ml. All submarine strains grew in the presence of NaCl at 3% or higher, but no strains from terrestrial hot springs would grow at concentrations higher than 1% NaCl. The growth rate of submarine
Thermus
strains was not stimulated by NaCl and was reduced at NaCl concentrations higher than 1%. The pattern of growth of these isolates on single carbon sources was similar to that of terrestrial isolates.
Occurrence of different types of infectious hematopoietic necrosis virus in fish
Authors: Hsu Y L , Engelking H M , Leong J C
Published: January 2020

The virion protein patterns of 71 isolates of infectious hematopoietic necrosis virus (IHNV) from the Pacific Northwest were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of [35S]-methionine-labeled virus. This analysis led to the classification of these virus isolates into four or more types. Type 1 virus was characterized by a nucleocapsid protein with an approximate molecular weight of 40,500. Type 2 and type 3 viruses have nucleocapsid proteins with molecular weights of 42,800 and 43,250, respectively. Type 2 virus was responsible for the recent epizootics of IHNV among fish in the lower Columbia River. The California IHNV isolates were type 3 with the exception of some of those isolated from fish at the Coleman Hatchery on the Sacramento River. These Coleman Hatchery isolates belonged to a type 4 virus group characterized by a larger glycoprotein of approximately 70,000 molecular weight. All other viruses examined had glycoproteins of 67,000 molecular weight. The "type 5" virus isolates were grouped together because they were not sufficiently distinct to warrant classification into a separate type. These findings have been useful in determining that a particular virus type is characteristic for a geographic area and will infect many different salmonid species in that area and the same type isolated from parental fish is responsible for the subsequent outbreak of the diseases in progeny.
Spatial and Temporal Variations in Bacterial Macromolecule Labeling with [
methyl
-
3
H]Thymidine in a Hypertrophic Lake
Authors: Robarts Richard D. , Wicks Richard J. , Sephton Lynne M.
Published: January 2020

The incorporation of [
methyl
-
3
H]thymidine into three macromolecular fractions, designated as DNA, RNA, and protein, by bacteria from Hartbeespoort Dam, South Africa, was measured over 1 year by acid-base hydrolysis procedures. Samples were collected at 10 m, which was at least 5 m beneath the euphotic zone. On four occasions, samples were concurrently collected at the surface. Approximately 80% of the label was incorporated into bacterial DNA in surface samples. At 10 m, total incorporation of label into bacterial macromolecules was correlated to bacterial utilization of glucose (
r
= 0.913,
n
= 13,
P
< 0.001). The labeling of DNA, which ranged between 0 and 78% of total macromolecule incorporation, was inversely related to glucose uptake (
r
= -0.823), total thymidine incorporation (
r
= -0.737), and euphotic zone algal production (
r
= -0.732,
n
= 13,
P
< 0.005). With decreased DNA labeling, increasing proportions of label were found in the RNA fraction and proteins. Enzymatic digestion followed by chromatographic separation of macromolecule fragments indicated that DNA and proteins were labeled while RNA was not. The RNA fraction may represent labeled lipids or other macromolecules or both. The data demonstrated a close coupling between phytoplankton production and heterotrophic bacterial activity in this hypertrophic lake but also confirmed the need for the routine extraction and purification of DNA during [
methyl
-
3
H]thymidine studies of aquatic bacterial production.
Occurrence and distribution of halophilic vibrios in subtropical coastal waters of Hong Kong
Authors: Chan K Y , Woo M L , Lo K W , French G L
Published: January 2020

The summer occurrence and distribution of halophilic vibrios in the subtropical coastal waters of Hong Kong were investigated. The density of vibrios in six sample sites ranged from 90 to 6,700 per ml, which made up 0.41 to 40% of the total bacterial populations of these sample sites. The sucrose-positive vibrios were found to be much more common (88% of total vibrios) than the sucrose-negative ones. A total of 48 strains belonging to six Vibrio species were fully characterized. Among these, Vibrio alginolyticus was the most frequently isolated, followed by V. parahaemolyticus, V. harveyi, V. vulnificus, V. campbellii, and V. fluvialis. The finding that eight of the nine strains of V. harveyi showed a positive Kanagawa reaction warrants further study.
Validity of Eucaryote Inhibitors for Assessing Production and Grazing Mortality of Marine Bacterioplankton
Authors: Taylor Gordon T. , Pace Michael L.
Published: January 2020

Application of eucaryote inhibitors to the estimation of production and grazing mortality of bacterioplankton was evaluated. Exposure to a range of concentrations of thiram, cycloheximide, and neutral red (0.4 to 210, 36 to 1,777, 4 to 346 μM, respectively) was 98 to 100% effective at inhibiting growth of a chrysomonad in culture. Exposure to colchicine and griseofulvin (50 to 1,000 μM for both) yielded only 24 to 94 and 53 to 79% inhibition, respectively. Exposures to thiram, neutral red, and griseofulvin were 90 to 100% effective at inhibiting growth in culture of a ciliate,
Cyclidium
sp., and the responses to colchicine and cycloheximide were variable (64 to 100 and 0 to 100% inhibition, respectively). Thiram and neutral red inhibited field populations of nanozooplankton more effectively than cycloheximide and colchicine. Direct effects of eucaryote inhibitors on growing cultures of bacterioplankton varied with parameters measured and duration of exposure. After 3-day exposures, specific growth rates and “instantaneous” heterotrophic potential ([
14
C]glucose uptake) were not consistently affected, but biosynthetic activity (RNA and DNA syntheses) was depressed. The degree of inhibition of isolates and field populations of phytoplankton depended upon type of inhibitor and phytoplankton species. In field experiments, it was possible to calculate rates of bacterioplankton production and grazing mortality for only 16 of 29 inhibitor experiments and for 4 of 10 size fractionation experiments. Bacterioplankton production and mortality estimates varied greatly with the eucaryote inhibitor used, and those derived from inhibition techniques were substantially different from those derived from fractionation techniques. The poor performances of both techniques are attributed to the following: (i) effects of inhibitors on phytoplankton, (ii) indirect effects of the inhibitors on bacterioplankton, and (iii) insufficient separation of grazers from prey by filtration techniques. Because of the inconsistent results obtained in this investigation, we strongly recommend exercising caution in the application of inhibitor techniques to ecological problems, especially in phototrophically dominated systems.
Solid-state 13C nuclear magnetic resonance spectroscopy of simultaneously metabolized acetate and phenol in a soil Pseudomonas sp
Authors: Heiman A S , Cooper W T
Published: January 2020

We investigated concentration-dependent primary and secondary substrate relationships in the simultaneous metabolism of the ubiquitous pollutant phenol and the naturally occurring substrate acetate by a Pseudomonas sp. soil isolate capable of utilizing either substance as a sole source of carbon and energy. In addition to conventional analytical techniques, solid-state 13C nuclear magnetic resonance spectroscopy was used to follow the cellular distribution of [1-13C]acetate in the presence of unlabeled phenol. With 5 mM acetate as the primary substrate, Pseudomonas sp. 9S8D2 removed 1 mM phenol (secondary substrate) at a rate of 2 nmol/mg of total cell protein. Although extensive acetate metabolism was indicated by a significant redistribution of the carboxyl label, this redistribution was not affected by the presence of phenol as a secondary substrate. When the primary and secondary substrate roles were reversed, however, the presence of 1 mM phenol altered the metabolism of 0.1 mM acetate, as evidenced by both the two- to fourfold increases in carboxyl label that appeared in terminal methyl and acyl chain methylene carbon resonances and the decrease in label that occurred in the carbohydrate spectral region. These results suggest that, when phenol is present as the primary substrate, acetate is preferentially shuttled into fatty acyl chain synthesis, whereas phenol carbon is funnelled into the tricarboxylic acid cycle. Thus, simultaneous use of a xenobiotic compound and a natural substrate apparently does occur, and the relative concentrations of the two substrates do influence the rate and manner in which the compounds are utilized.(ABSTRACT TRUNCATED AT 250 WORDS)
Dynamics of extracellular DNA in the marine environment
Authors: Paul J H , Jeffrey W H , DeFlaun M F
Published: January 2020

The production and turnover of dissolved DNA in subtropical estuarine and oligotrophic oceanic environments were investigated. Actively growing heterotrophic bacterioplankton (i.e., those capable of [3H]thymidine incorporation) were found to produce dissolved DNA, presumably through the processes of death and lysis, grazing by bacteriovores, and excretion. Production of dissolved DNA as determined by [3H]thymidine incorporation was less than or equal to 4% of the ambient dissolved DNA concentration per day. In turnover studies, the addition of [3H]DNA (Escherichia coli chromosomal) to seawater resulted in rapid hydrolysis and uptake or radioactivity by microbial populations. DNA was hydrolyzed by both cell-associated and extracellular nucleases, in both estuarine and offshore environments. Kinetic analysis performed for a eutrophic estuary indicated a turnover time for dissolved DNA as short as 6.5 h. Microautoradiographic studies of bacterial populations in Tampa Bay indicated that filamentous and attached bacteria took up most of the radioactivity from [3H]DNA. Dissolved DNA is therefore a dynamic component of the dissolved organic matter in the marine environment, and bacterioplankton play a key role in the cycling of this material.
High incidence of selenite-resistant bacteria from a site polluted with selenium
Authors: Burton G A , Giddings T H , DeBrine P , Fall R
Published: January 2020

The level of selenium-resistant bacteria in water, algal mats, and sediment from Kesterson reservoir, Calif., a site with known selenium pollution, was compared with that in nearby Volta reservoir, a site with low selenium levels. A high percentage (greater than 50%) of all isolates from the Kesterson samples were resistant to 10 mM selenite. In contrast, only a small percentage of the Volta isolates were resistant to this level of selenite. The identity of some selenite-resistant isolates and MICs of selenite, selenate, arsenate, tellurite, and tellurate were determined.
Arginine deiminase system and bacterial adaptation to acid environments
Authors: Marquis R E , Bender G R , Murray D R , Wong A
Published: January 2020

The arginine deiminase system in a variety of streptococci and in Pseudomonas aeruginosa was found to be unusually acid tolerant in that arginolysis occurred at pH values well below the minima for growth and glycolysis. The acid tolerance of the system allowed bacteria to survive potentially lethal acidification through production of ammonia to raise the environmental pH value.
Nitrosubstituted Aromatic Compounds as Nitrogen Source for Bacteria
Authors: Bruhn C. , Lenke H. , Knackmuss H.-J.
Published: January 2020

Bacteria which utilized nitroaromatic compounds (0.5 mM) as sole source of nitrogen were isolated from soil. With 2,6-dinitrophenol and succinate as carbon source, a
Pseudomonas
strain was isolated which liberated and assimilated nitrite. Approximately 2 mol of NO
2
−
per mol of 2,6-dinitrophenol was released by resting cells. The xenobiotic compound was totally degraded, although specific growth yields were low even with succinate as a carbon source.
Specific and sensitive plate assay for bacterial lipases
Authors: Kouker G , Jaeger K E
Published: January 2020

A plate assay to detect bacterial lipase (EC 3.1.1.3) in a medium containing trioleoylglycerol and the fluorescent dye rhodamine B is presented. Substrate hydrolysis causes the formation of orange fluorescent halos around bacterial colonies visible upon UV irradiation. The logarithm of lipase activity from cell-free culture supernatants is linearly correlated with the diameter of halos, thereby allowing quantitation of lipase activities ranging from 1 to 30 nkat.
Biofilm Dynamics and Kinetics during High-Rate Sulfate Reduction under Anaerobic Conditions
Authors: Nielsen Per Halkjaer
Published: January 2020

The sulfate kinetics in an anaerobic, sulfate-reducing biofilm were investigated with an annular biofilm reactor. Biofilm growth, sulfide production, and kinetic constants (
K

m

and
V
max
) for the bacterial sulfate uptake within the biofilm were determined. These parameters were used to model the biofilm kinetics, and the experimental results were in good agreement with the model predictions. Typical zero-order volume rate constants for sulfate reduction in a biofilm without substrate limitation ranged from 56 to 93 μmol of SO
2
4
-cm
−3
h
−1
at 20�C. The temperature dependence (Q
10
) of sulfate reduction was equivalent to 3.4 at between 9 and 20�C. The measured rates of sulfate reduction could explain the relatively high sulfide levels found in sewers and wastewater treatment systems. Furthermore, it has been shown that sulfate reduction in biofilms just a few hundred micrometers thick is limited by sulfate diffusion into biofilm at concentrations below 0.5 mM. This observation might, in some cases, be an explanation for the relatively poor capacity of the sulfate-reducing bacteria to compete with the methanogenic bacteria in anaerobic wastewater treatment in submerged filters.
Involvement of the spore coat in germination of Bacillus cereus T spores
Authors: Kutima P M , Foegeding P M
Published: January 2020

Bacillus cereus T spores were prepared on fortified nutrient agar, and the spore coat and outer membrane were extracted by 0.5% sodium dodecyl sulfate-100 mM dithiothreitol in 0.1 M sodium chloride (SDS-DTT) at pH 10.5 (coat-defective spores). Coat-defective spores in L-alanine plus adenosine germinated slowly and to a lesser extent than spores not treated with SDS-DTT, as determined by decrease in absorbance and release of dipicolinic acid and Ca2+. Spores germinated in calcium dipicolinate only after treatment with SDS-DTT. Biphasic and triphasic germination kinetics were observed with normal and coat-defective spores, respectively, in an environment with temperature increasing from 20 to 65 degrees C at a rate of 1 degree C/min. Therefore, the physical and biochemical processes involved in germination are modified by coat removal. The data suggest that a portion of the germination apparatus located interior to the coat may be protected by the coat and outer membrane or that the coat and outer membrane otherwise enhance germination in L-alanine plus adenosine. When coat-defective spores were heat activated with the dialyzed (12,000-Mr cutoff) components extracted from the spores, germination of the SDS-DTT-treated spores was enhanced; thus, one or more components located in the spore coat or outer membrane with a molecular weight greater than 12,000 were essential for fast germination.
Ecophysiological adaptations of anaerobic bacteria to low pH: analysis of anaerobic digestion in acidic bog sediments
Authors: Goodwin S , Zeikus J G
Published: January 2020

The dynamics of anaerobic digestion were examined in the low-pH sediments of Crystal Bog in Wisconsin. The sediments (pH 4.9) contained 71% organic matter and the following concentrations of dissolved gases (micromoles per liter): CO2, 1,140; CH4, 490; and H2, 0.01. The rate of methane production was 6.2 mumol/liter of sediment per h, which is slower than eutrophic, neutral sediments. Microbial metabolic processes displayed the following pH optima: hydrolysis reactions, between 4.2 and 5.6; aceticlastic methanogenesis, 5.2; and hydrogen-consuming reactions, 5.6. The turnover rate constants for key intermediary metabolites were (h-1): glucose, 1.10; lactate, 0.277; acetate, 0.118; and ethanol, 0.089. The populations of anaerobes were low, with hydrolytic groups (10(6)/ml) several orders of magnitude higher than methanogens (10(2)/ml). The addition of carbon electron donors to the sediment resulted in the accumulation of hydrogen, whereas the addition of hydrogen resulted in the accumulation of fatty acids and the inhibition of hydrogen-producing acetogenic reactions. Strains of Lactobacillus, Clostridium, and Sarcina ventriculi were isolated from the bog, and their physiological attributes were characterized in relation to hydrolytic process functions in the sediments. The present studies provide evidence that the pH present in the bog sediments alter anaerobic digestion processes so that total biocatalytic activity is lower but the general carbon and electron flow pathways are similar to those of neutral anoxic sediments.
Development of High-Frequency Delivery System for Transposon Tn
919
in Lactic Streptococci: Random Insertion in
Streptococcus lactis
subsp.
diacetylactis
18-16
Authors: Hill Colin , Daly Charles , Fitzgerald Gerald F.
Published: January 2020

The conjugative transposon Tn
919
, originally isolated in
Streptococcus sanguis
FC1, is capable of low-frequency transfer (10
−7
and 10
−8
per recipient) on membrane filters to a wide number of streptococcal recipients including the industrially important lactic streptococci. The introduction of pMG600 (Lac
+
Lax
−
; a lactose plasmid capable of conjugative transfer at high frequencies and which, in certain hosts, confers an unusual clumping phenotype) into a
Streptococcus lactis
CH919 donor, generating
S. lactis
CH001, resulted in a significant improvement in the transfer frequency of Tn
919
to
S. lactis
CK50 (1.25 � 10
−4
per recipient). In addition, these matings could be performed on agar surfaces, allowing the recovery of a greater number of recipients than with filter matings. Tn
919
also transferred at high frequency to
S. lactis
subsp.
diacetylactis
18-16S but not to
Streptococcus cremoris
strains. Insertion in 18-16S transconjugants generated from filter matings with an
S. lactis
CH919 donor was random, occurring at different sites on the chromosome and also in plasmid DNA. Thus, the conditions necessary for the practical exploitation of Tn
919
in the targeting and cloning of genes from a member of the lactic streptococci, namely, high-frequency delivery and random insertion in host DNA, were achieved.
Modification of membrane sulfhydryl groups in bacteriostatic action of nitrite
Authors: Buchman G W , Hansen J N
Published: January 2020

The mechanism by which nitrite inhibits outgrowing spores of Bacillus cereus T was examined by using techniques developed earlier for nitrite analogs. The morphological stage of inhibition, cooperativity effects, effect of pH on inhibition, kinetics of protection against iodoacetate incorporation into membrane sulfhydryl groups, and protection against the bacteriocidal effect of carboxymethylation by iodoacetate indicate that nitrite acts as a membrane-directed sulfhydryl agent. The mechanism by which nitrite modifies the chemical reactivity of the sulfhydryl group could be either direct covalent modification or inactivation through communication with another modified membrane component. Profiles of pH effects suggest that the active agent is the protonated form of nitrite. The nitrite concentrations which modify membrane sulfhydryl activity coincide with those which have a bacteriostatic effect. These results are consistent with membrane sulfhydryl modification as a component of the mechanism of nitrite-induced bacteriostasis in this aerobic sporeformer.
Temperature Characteristics of Photosynthetic and Heterotrophic Activities: Seasonal Variations in Temperate Microbial Plankton
Authors: Li W. K. W. , Dickie P. M.
Published: January 2020

The seasonal variation in temperature characteristics of photosynthetic and heterotrophic activities in the microbial plankton of Bedford Basin, Nova Scotia, was investigated. Measurements were made of the photosynthetic uptake of [
14
C]bicarbonate and its incorporation into cellular protein as well as the heterotrophic uptake of
3
H-labeled amino acids and their incorporation into cellular protein. Activity-temperature curves were analyzed objectively by nonlinear estimation of parameters from various mathematical models. Over the seasonal cycle, the cardinal temperatures and a parameter formally equivalent to the thermodynamic enthalpy of activation for most of the four processes measured were positively correlated with the water temperature. The temperature sensitivity of metabolic activity (i.e., change in activity per unit change in temperature) was indexed by the tangent to the activity-temperature curves. When this index was expressed in dimensionless form by normalization to the scaling factor of the activity-temperature curves, the resulting relative temperature sensitivity, evaluated at the prevailing temperature, proved to be statistically invariant throughout the year. During the height of the spring bloom, the water temperature (−0.3�C) was not so low as to inhibit metabolic activity of either the phytoplankton or the bacterioplankton. The evidence suggests that heterotrophic utilization of products is not suppressed during the spring phytoplankton bloom.
Purification and Characterization of a Substrate-Size-Recognizing Metalloendopeptidase from
Streptococcus cremoris
H61
Authors: Yan Tsong-Rong , Azuma Norihiro , Kaminogawa Shuichi , Yamauchi Kunio
Published: January 2020

During the ripening of Gouda-type cheese, two kinds of endopeptidases were found to participate in the degradation of αs1-CN(f1-23), a specific product from αs1-casein hydrolyzed by chymosin. One of the endopeptidases, lactic acid bacteria endopeptidase (LEP-II), which can recognize the size of its substrates, has already been purified and characterized (T. R. Yan, N. Azuma, S. Kaminogawa, and K. Yamauchi, Eur. J. Biochem. 163:259-265, 1987). The other endopeptidase, LEP-I, was purified to homogeneity by conventional chromatographic techniques from
Streptococcus cremoris
H61. The enzyme appeared to be monomeric, with an apparent molecular weight of 98,000, and its isoelectric point was 5.1. For the hydrolysis of αs1-CN(f1-23), the enzyme had an optimum pH and temperature of 7.0 to 7.5 and 40�C, respectively. Its activity was inhibited by such chelating agents as EDTA and 1,10-phenanthrolin, and it could be fully reactivated by Mn
2+
. Inhibitors specific for serine and thiol proteases had no effect on the protease activity. The enzyme showed a high affinity toward the Glu-Asn peptide bond of αs1-CN(f1-23) and αs1-CN(f91-100) but showed no hydrolysis activity toward αs1-CN(f1-52), αs1-CN(61-122), αs1-CN(136-196), αs1-casein, β-casein, κ-casein, α-lactalbumin, and β-lactoglobulin. The
K

m

and
V
max
of LEP-I for αs1-CN(f1-23) were 14.2 pM and 139 U, respectively.
Sodium bicarbonate reduces viability and alters aflatoxin distribution of Aspergillus parasiticus in Czapek's agar
Authors: Montville T J , Goldstein P K
Published: January 2020

The potential of sodium bicarbonate to inhibit growth of and aflatoxin synthesis by Aspergillus parasiticus was examined in Czapek's agar (CA), a medium in which fluorescence under UV light indicates aflatoxin production. Incorporation of sodium bicarbonate (SB) into CA at 0.011, 0.022, and 0.033 mol% reduced cell viability 63-, 10(3)-, and greater than 10(7)-fold, respectively. Colonies resulting from surviving cells did not fluoresce under UV light, but thin-layer chromatography analysis of culture extracts detected aflatoxins. Potassium bicarbonate (KB) at 0.011 and 0.022 mol% produced inhibitory effects similar to those of SB, but NaCl and silica had no effect. After 7 days, control cultures had the normal aflatoxin distribution (B1 greater than G1 greater than B2 greater than G2), but this distribution shifted to B2 greater than B1 approximately equal to G2 greater than G1 during prolonged incubation. Cultures supplemented with SB and KB contained mostly aflatoxins B1 and G1 after 28 days. Both SB and KB raised the pH of CA to 7.5 to 8.5 at the time of growth. Culture growth on CA adjusted to pH 7.5 to 8.5 with NaOH was not inhibited but exhibited reduced fluorescence and elevated levels of aflatoxins B1 and G1. Thus, while bicarbonate inhibition of growth could not be attributed to pH elevation, the lack of culture fluorescence on CA-SB and CA-KB and the altered aflatoxin distribution were caused by the ability of SB and KB to elevate pH.
Isolation and Characterization of
Frankia
sp. Strain FaC1 Genes Involved in Nitrogen Fixation
Authors: Ligon James M. , Nakas James P.
Published: January 2020

Genomic DNA was isolated from
Frankia
sp. strain FaC1, an
Alnus
root nodule endophyte, and used to construct a genomic library in the cosmid vector pHC79. The genomic library was screened by in situ colony hybridization to identify clones of
Frankia
nitrogenase (
nif
) genes based on DNA sequence homology to structural nitrogenase genes from
Klebsiella pneumoniae.
Several
Frankia nif
clones were isolated, and hybridization with individual structural nitrogenase gene fragments (
nifH, nifD
, and
nifK
) from
K. pneumoniae
revealed that they all contain the
nifD
and
nifK
genes, but lack the
nifH
gene. Restriction endonuclease mapping of the
nifD
and
nifK
hybridizing region from one clone revealed that the
nifD
and
nifK
genes in
Frankia
sp. are contiguous, while the
nifH
gene is absent from a large region of DNA on either side of the
nifDK
gene cluster. Additional hybridizations with gene fragments derived from
K. pneumoniae
as probes and containing other genes involved in nitrogen fixation demonstrated that the
Frankia nifE
and
nifN
genes, which play a role in the biosynthesis of the iron-molybdenum cofactor, are located adjacent to the
nifDK
gene cluster.
Effects of potassium ion concentrations on the antimicrobial activities of ionophores against ruminal anaerobes
Authors: Dawson K A , Boling J A
Published: January 2020

The antimicrobial activities of monensin and lasalocid against representative strains of ruminal bacteria were evaluated in medium containing three different concentrations of potassium (1.3, 7.9, or 23.3 mM). The growth of Eubacterium ruminantium was inhibited by low concentrations of ionophores (less than or equal to 0.16 mg/liter), while the strain of Streptococcus bovis tested was resistant to high concentrations of ionophores (40 mg/liter) at all potassium concentrations tested. The MICs of the ionophores for strains of Bacteroides succinogenes, Butyrivibrio fibrisolvens, Ruminococcus albus, and Ruminococcus flavefaciens and for one strain of Bacteroides ruminicola increased with increasing potassium concentrations in the medium. High concentrations of ionophores (40 mg/liter) decreased the maximum cell yields or increased the lag times or both in cultures of one strain of Bacteroides ruminicola and two strains of Selenomonas ruminantium but did not completely inhibit the growth of these organisms. Increased potassium concentrations in the medium (from 7.9 to 23.3 mM) decreased the lag times or increased the cell yields or both when these three strains were grown in ionophore-containing medium, while the activities of lasalocid and monensin against these organisms were enhanced in the medium containing low potassium concentrations (1.3 mM). The data from this study suggest that extracellular potassium concentrations may influence the antimicrobial activities of ionophores in the rumen.
Microbial Growth Rates and Biomass Production in a Marine Sediment: Evidence for a Very Active but Mostly Nongrowing Community
Authors: Novitsky James A.
Published: January 2020

Biomass, nucleic acid synthesis, and specific growth rates of the microbial communities were measured throughout a vertical profile of a coastal marine sediment. The microbial biomass, as determined by ATP concentrations, in the sediment-water interface was over twice that measured in the other horizons of a 10-cm profile. Likewise, biomass carbon production, as determined by DNA synthesis, and the specific growth rate, as determined from the kinetics of [
3
H]ATP pool labeling, were also elevated at the interface. These results indicate that, due to a large and active community in the interface, the greatest amount of microbial activity, growth, and biosynthesis occurs within the first few millimeters of sediment. These results notwithstanding, a combination of two independent techniques established that over 90% of the sediment-water interface community was not actively growing.