bluuebunny/crossref_metadata_2025 huggingface dataset:
Chemical transformation of eremofortin C into PR toxin
Authors: Li S Y , Chang S C , Wei R D
Published: January 2020

The natural products of both eremofortin C (EC) and PR toxin are secondary metabolites of Penicillium roqueforti. Because the chemical structures of EC and PR toxin are closely related to each other and differ only by a hydroxyl functional group in EC and an aldehyde functional group in PR toxin at the C-12 position, the chemical transformation of EC into PR toxin was investigated. Oxidation with a chromic anhydride-pyridine complex was found to be the most satisfactory method.
Glutamine synthetase activity in the ruminal bacterium Succinivibrio dextrinosolvens
Authors: Patterson J A , Hespell R B
Published: January 2020

Succinivibrio dextrinosolvens C18 was found to possess glutamine synthetase (GS), urease, glutamate dehydrogenase, and several other nitrogen assimilation enzymes. When grown in continuous culture under ammonia limitation, both GS and urease activities were high and glutamate dehydrogenase activity was low, but the opposite activity pattern was observed for growth in the presence of ample ammonia. The addition of high-level (15 mM) ammonium chloride to ammonia-limited cultures resulted in a rapid loss of GS activity as measured by either the gamma-glutamyl transferase or forward assay method with cells or extracts. No similar activity losses occurred for urease, glutamate dehydrogenase, or pyruvate kinase. The GS activity loss was not prevented by the addition of chloramphenicol and rifampin. The GS activity could be recovered by washing or incubating cells in buffer or by the addition of snake venom phosphodiesterase to cell extracts. Manganese inhibited the GS activity (forward assay) of untreated cells but stimulated the GS activity in ammonia-treated cells. Alanine, glycine, and possibly serine were inhibitory to GS activity. Optimal pH values for GS activity were 7.3 and 7.4 for the forward and gamma-glutamyl transferase assays, respectively. The glutamate dehydrogenase activity was NADPH linked and optimal in the presence of KCl. The data are consistent with an adenylylation-deadenylylation control mechanism for GS activity in S. dextrinosolvens, and the GS pathway is a major route for ammonia assimilation under low environmental ammonia levels. The rapid regulation of the ATP-requiring GS activity may be of ecological importance to this strictly anaerobic ruminal bacterium.
Improvement of a Wine
Saccharomyces cerevisiae
Strain by a Breeding Program
Authors: Romano Patrizia , Soli M. Grazia , Suzzi Giovanna , Grazia Luigi , Zambonelli Carlo
Published: January 2020

Hybridization by spore conjugation was used to develop new and improved wine yeasts of
Saccharomyces cerevisiae.
The procedure was achieved with diploid, homothallic strains with high sporulation frequency and high spore viability. The method was verified by crossing flocculent and non-H
2
S-forming strains. Single-spore descendants of the hybrids were studied by tetrad analysis with regard to the aforementioned characters and the other two winemaking traits, i.e., ethanol production and fermentation rate. A highly flocculent, non-H
2
S-forming wine yeast strain with a high fermentation rate and high ethanol production was obtained.
Selective and differential medium for detecting Clostridium botulinum
Authors: Silas J C , Carpenter J A , Hamdy M K , Harrison M A
Published: January 2020

A selective and differential growth medium was developed for detection of Clostridium botulinum types A, B, and F. The medium consisted of peptone-glucose-yeast extract agar supplemented with cycloserine, 250 micrograms/ml; sulfamethoxazole, 76 micrograms/ml; and trimethoprim, 4 micrograms/ml as selective inhibitors and various types and levels of botulinal antibodies for type differentiation in the immunodiffusion reaction. Growth of proteolytic types of C. botulinum were not affected by the incorporation of the selective agents, but some nonproteolytic types were suppressed. Cross-reactions between types A and B were visually distinguishable, whereas cross-reactions between type F and Clostridium sporogenes did not occur at the optimum antibody titer. Optimum antibody titer varied with toxin type. The proposed selective differential medium should be valuable in isolating and typing of proteolytic C. botulinum types A, B, and F from samples containing mixed microbial populations.
Antibiotic resistance and population structure in Escherichia coli from free-ranging African yellow baboons
Authors: Routman E , Miller R D , Phillips-Conroy J , Hartl D L
Published: January 2020

Two collections of Escherichia coli from human hosts and one from free-ranging African yellow baboons were examined for the ability to utilize various sugars (biotype) and for resistance to antibiotics. The frequency of antibiotic resistance in the E. coli flora of baboons that feed regularly in village garbage dumps was found to be no greater than that in baboons not associated with human habitation. The frequency of antibiotic resistance in E. coli isolated from baboons is similar to that in E. coli isolated from humans before the widespread use of antibiotics but significantly lower than that in recent isolates from humans. The biotype data indicate that the amount and distribution of genetic variation in the E. coli among free-ranging baboon troops are similar to those in isolates from humans. However, E. coli isolates from baboons are able to utilize a greater variety of sugars as their sole carbon source, possibly because of a greater variety of sugars in the baboon diet.
Habitat Segregation and Biochemical Activities of Marine Members of the Family
Vibrionaceae
Authors: Simidu Usio , Tsukamoto Kumiko
Published: January 2020

A comparative study of marine members of the family
Vibrionaceae
with the technique of numerical taxonomy revealed habitat segregation as well as a cosmopolitan nature of species distribution among the vibrios in different marine environments. The bacterial strains analyzed were isolated from seawater, sediments, phyto- and zooplankton, and fish in the Indian Ocean, the South and East China Sea, and West Pacific Ocean, and coastal areas of Japan. A total of 155 morphological, physiological, and biochemical tests were carried out for each of 405 strains examined. The results showed that most of the large taxonomical clusters which emerged from the computation corresponded to ecological groups which have particular niches. For instance, each group of seawater vibrios inhabited a particular water layer of limited depth range, in spite of the fact that strains of the group were isolated from sampling locations spread over a wide area from the Indian Ocean to Japanese coast. Various vibrio groups showed remarkable differences in their physiological and biochemical activities, and the activities of each group seemed to correspond with its ecological niche. The strains which inhabited surface-water layers grew fast and actively utilized many high-molecular-weight organic compounds and carbohydrates that are derived from fresh, easily degradable organic matter present in the surface waters, whereas the middle- and deep-water vibrios did not decompose most of the high-molecular-weight organic compounds except chitin but, rather, utilized some carbohydrates and organic acids which seemed to be derived from refractory particulate organic matter present in the deeper waters.
Denitrification Rates in the Low-Oxygen Waters of the Stratified Baltic Proper
Authors: R�nner Ulf , S�rensson Fred
Published: January 2020

Denitrification activity was shown in the deep, low-oxygen waters of the Baltic proper by both in vitro and in situ methods. The vertical distribution of NO
3
−
in the water column showed nitrate consumption and NO
2
−
and N
2
O maxima in the deep waters when O
2
was below 0.2 ml liter
−1
, which is suggestive evidence for denitrification. Direct in situ evidence for denitrification was obtained by finding an N
2
saturation of up to 108% in the deep waters. When these waters were incubated with
15
NO
3
−
,
15
N
2
was produced. Quantification of the denitrification rate done by the addition of C
2
H
2
to water samples from the active depths showed a rate of about 0.10 μmol liter
−1
day
−1
.
Dissimilatory nitrate reduction to nitrate, nitrous oxide, and ammonium by Pseudomonas putrefaciens
Authors: Samuelsson M O
Published: January 2020

The influence of redox potential on dissimilatory nitrate reduction to ammonium was investigated on a marine bacterium, Pseudomonas putrefaciens. Nitrate was consumed (3.1 mmol liter-1), and ammonium was produced in cultures with glucose and without sodium thioglycolate. When sodium thioglycolate was added, nitrate was consumed at a lower rate (1.1 mmol liter-1), and no significant amounts of nitrite or ammonium were produced. No growth was detected in glucose media either with or without sodium thioglycolate. When grown on tryptic soy broth, the production of nitrous oxide paralleled growth. In the same medium, but with sodium thioglycolate, nitrous oxide was first produced during growth and then consumed. Acetylene caused the nitrous oxide to accumulate. These results and the mass balance calculations for different nitrogen components indicate that P. putrefaciens has the capacity to dissimilate nitrate to ammonium as well as to dinitrogen gas and nitrous oxide (denitrification). The dissimilatory pathway to ammonium dominates except when sodium thioglycolate is added to the medium.
Streptococcus cremoris M12R transconjugants carrying the conjugal plasmid pTR2030 are insensitive to attack by lytic bacteriophages
Authors: Steenson L R , Klaenhammer T R
Published: January 2020

Conjugal transfer of lactose-fermenting ability (Lac+), nisin resistance (Nisr), and phage resistance (Hsp+) was demonstrated in matings between Streptococcus lactis ME2 (donor) and Streptococcus cremoris M43a (recipient), a derivative of M12R. Transconjugants were detected by transfer of Lac+ and were found to exhibit Nisr and harbor a 40-megadalton plasmid (pTR1040). Fifty-six percent of Lac+ transconjugants were resistant to the S. cremoris M12R lytic phage. Efficiency of plaquing for phage m12r . M12 on a phage-resistant transconjugant, T2r-M43a, was less than 4.3 X 10(-10). Five additional phages which were virulent for S. cremoris M12R and isolated from industrial sources failed to plaque on S. cremoris T2r-M43a. Mating experiments with T2r-M43a revealed that phage resistance was accompanied by high-frequency conjugation ability (Tra+) and the appearance of both pTR1040 and pTR2030 encoding Lac+ Nisr and Tra+ Hsp+, respectively, in transconjugants of S. lactis LM2302. Phage-sensitive Lac+ transconjugants of S. cremoris M43a (T2s-M43a) showed no conjugal ability. These observations confirmed that pTR2030 was present and responsible for the phage resistance and conjugal ability exhibited by the S. cremoris transconjugant T2r-M43a. Unlike the S. lactis LM2302 transconjugant carrying pTR2030, resistance of T2r-M43a to phage was not affected at high temperatures (35 to 40 degrees C) or destabilized in repeated transfers through a starter culture activity test. These results demonstrated that phage resistance conferred by pTR2030 in the S. cremoris transconjugant was effective against industrially significant phages under fermentation conditions normally encountered during cheese manufacture.
Effect of temperature on bacterial species diversity in thermophilic solid-waste composting
Authors: Strom P F
Published: January 2020

Continuously thermophilic composting was examined with a 4.5-liter reactor placed in an incubator maintained at representative temperatures. Feed was a mixture of dried table scraps and shredded newspaper wetted to 55% moisture. One run at 49 degrees C (run A) employed a 1:4 feed-to-compost ratio, while the other runs used a 10:1 ratio and were incubated at 50, 55, 60, or 65 degrees C. Due to self-heating, internal temperatures of the composting mass were 0 to 7 degrees C hotter than the incubator. Two full-scale composting plants (at Altoona, Pa., and Leicester, England) were also examined. Plate counts per gram (dry weight) on Trypticase soy broth (BBL Microbiology Systems) with 2% agar ranged from 0.7 X 10(9) to 5.3 X 10(9) for laboratory composting and 0.02 X 10(9) to 7.4 X 10(9) for field composting. Fifteen taxa were isolated, including 10 of genus Bacillus, which dominated all samples except that from run A. Species diversity decreased markedly in laboratory composting at 60 degrees C and above, but was similar for the three runs incubated at 49, 50, and 55 degrees C. The maximum desirable composting temperature based on species diversity is thus 60 degrees C, the same as that previously recommended based on measures of the rate of decomposition.
Protein synthesis by mammalian cells treated with C-3-modified analogs of the 12,13-epoxytrichothecenes T-2 and T-2 tetraol
Authors: Ehrlich K C , Daigle K W
Published: January 2020

Modification at the C-3 position of the trichothecenes T-2 and T-2 tetraol affected their ability to inhibit protein synthesis in African green monkey kidney (Vero) and mouse erythroleukemia cells. Replacement of the 3-hydroxyl of T-2 with hydrogen caused a 24-fold decrease in activity, whereas acetylation resulted in a 500-to 1,000-fold decrease. Protection of the 3-hydroxyl with a tetrahydropyranyl moiety gave an analog that was 37-fold more inhibitory to Vero than to mouse erythroleukemia cells; with the other analogs a similar effect on protein synthesis was found for both types of cells. The analogs obtained after alkaline hydrolysis were much less potent than the parent trichothecenes. The 3-tetrahydropyranyl-modified analog was equivalent in potency to T-2 tetraol, while the deoxygenated species was at least threefold less potent. All T-2 analogs caused some degree of polysome "runoff," thereby demonstrating that these species inhibit protein synthesis at the chain initiation stage when added at their 50% infective dose concentrations or lower. From these results, we suggest that the 3-hydroxyl moiety is essential for T-2 to exhibit such high activity on eucaryotic cell protein synthesis and that modification at the C-3 position decreases but does not eliminate this activity.
Thermophilic Anaerobic Biodegradation of [
14
C]Lignin, [
14
C]Cellulose, and [
14
C]Lignocellulose Preparations
Authors: Benner Ronald , Hodson Robert E.
Published: January 2020

Thermophilic (55�C) anaerobic enrichment cultures were incubated with [
14
C-lignin]lignocellulose, [
14
C-polysaccharide]lignocellulose, and kraft [
14
C]lignin prepared from slash pine,
Pinus elliottii
, and
14
C-labeled preparations of synthetic lignin and purified cellulose. Significant but low percentages (2 to 4%) of synthetic and natural pine lignin were recovered as labeled methane and carbon dioxide during 60-day incubations, whereas much greater percentages (13 to 23%) of kraft lignin were recovered as gaseous end products. Percentages of label recovered from lignin-labeled substrates as dissolved degradation products were approximately equal to percentages recovered as gaseous end products. High-pressure liquid chromatographic analyses of CuO oxidation products of sound and degraded pine lignin indicated that no substantial chemical modifications of the remaining lignin polymer, such as demethoxylation and dearomatization, occurred during biodegradation. The polysaccharide components of pine lignocellulose and purified cellulose were relatively rapidly mineralized to methane and carbon dioxide; 31 to 37% of the pine polysaccharides and 56 to 63% of the purified cellulose were recovered as labeled gaseous end products. An additional 10 to 20% of the polysaccharide substrates was recovered as dissolved degradation products. Overall, these results indicate that elevated temperatures can greatly enhance rates of anaerobic degradation of lignin and lignified substrates to methane and low-molecular-weight aromatic compounds.
Micro-lipid-droplet encapsulation of Bacillus thuringiensis subsp. israelensis delta-endotoxin for control of mosquito larvae
Authors: Cheung P Y , Hammock B D
Published: January 2020

The crystal delta-endotoxin of Bacillus thuringiensis subsp. israelensis is less toxic to larvae of Anopheles freeborni than to larvae of Aedes aegypti. However, when solubilized crystal was used, larvae from both species showed similar sensitivities. This effect presumably was due to the differences in feeding behavior between the two mosquito larvae when crystal preparations are used. A procedure is described whereby both crystal and solubilized B. thuringiensis subsp. israelensis toxin were emulsified with Freund incomplete adjuvant, with retention of toxicity. The use of Freund incomplete adjuvant also allowed one to assay the solubilized toxin at a low nanogram level. Furthermore, coating the toxin with lipophilic material altered the buoyancy of the toxin and reversed the sensitivities of the two mosquito larvae toward the B. thuringiensis subsp. israelensis toxin. This difference in buoyancy was determined by using an enzyme-linked immunosorbent assay that was specific for the toxic peptides. These data indicate that economically feasible buoyant formulations for the B. thuringiensis subsp. israelensis crystal can be developed.
Reactions of fish to microorganisms in wastewater
Authors: Buras N , Duek L , Niv S
Published: January 2020

Fish were inoculated with various microorganisms present in wastewater. A threshold concentration was determined over which these microorganisms were recovered from the muscles. The threshold concentrations were different for bacteria, bacteriophages, and polio 1 LSc virus. The threshold values were lower when fish were inoculated than when they were immersed in water containing these organisms. Depuration experiments were efficient when the fish did not contain high concentrations of bacteria in their muscles. As the threshold concentrations are an expression of the capability of the immune system of the fish, these values can be useful for the design and management of fishponds in which treated wastewater is used.
Bioreactor for the study of defined interactions of toxic metals and biofilms
Authors: Hsieh K M , Lion L W , Shuler M L
Published: January 2020

A novel bioreactor system constructed for studies of the interactions of heavy metals and microbial cells at the solid-solution interface is described. The applicability of this experimental system to meet the severe constraints imposed on such an apparatus by the requirements for an unambiguous interpretation of data and for mathematical modeling of these interactions was explored with the trace metal lead and with the marine bacterium Pseudomonas atlantica. A chemically defined medium composed of the major components of seawater, simple salts required for growth, glucose, and the single amino acid glycine was derived. It supported a maximum growth rate several times less than that in a complex medium, but provided growth to high cell densities and the formation of biopolymer and supported the development of a monolayer biofilm. The use of such a medium in conjunction with our bioreactor system minimized trace metal contamination while allowing quantification of the partitioning of lead onto various reactor surfaces. Lead adsorption by reactor walls and model surfaces was linear with equilibrium led concentration up to 6 X 10(-6) mol/liter. Equilibrium lead adsorption due to P. atlantica biofilm surfaces ranged from 20 to 40% at a total lead concentration of 10(-6) mol/liter depending upon solution pH and ionic composition, indicating that biofilms can play an important role in controlling toxic metal concentrations in natural systems.
X-ray microanalytic method for measurement of dry matter and elemental content of individual bacteria
Authors: Heldal M , Norland S , Tumyr O
Published: January 2020

A method for the determination of dry matter and elemental content of individual bacterial cells is described. The method is based on energy-dispersive X-ray microanalysis in a transmission electron microscope. A theory for area correction of intensity is developed. Escherichia coli in the late exponential phase of growth and early stationary phase (glucose limited) had an average dry matter content of 278 and 154 fg/cell, respectively. Of the elements detected, sodium, magnesium, phosphorus, sulphur, chlorine, potassium, and calcium together made up 15 to 17% of the dry matter content. A phosphorus content of 4.2 to 5.4% of the dry matter was found in these cells. Volume measurements of air-dried cells gave an average of 1.20 to 1.25 micron3. These results emphasize that dry matter content and elemental composition can be measured directly on single cells from complex microbial communities.
Inhibitory effect of a copper-dipeptide complex on the establishment of a Clostridium perenne strain in the intestinal tract of gnotobiotic mice
Authors: Dubos F , Pelissier J P , Andrieux C , Ducluzeau R , Raibaud P
Published: January 2020

A semisynthetic diet fed to axenic mice was found to prevent the establishment of a Clostridium perenne strain in their intestinal tract. This inhibitory effect did not occur when axenic mice were preinoculated with a strain of Clostridium difficile. The inhibitory effect was related to the presence in the intestinal contents of axenic mice of both dietary copper and a dipeptide, aspartic-epsilon-lysine. When C. difficile was inoculated into axenic mice, the dipeptide disappeared from the digesta, and C. perenne became established even in the presence of high concentrations of copper.
Effect of Fall Turnover on Terminal Carbon Metabolism in Lake Mendota Sediments
Authors: Phelps T. J. , Zeikus J. G.
Published: January 2020

The carbon and electron flow pathways and the bacterial populations responsible for the transformation of H
2
-CO
2
, formate, methanol, methylamine, acetate, ethanol, and lactate were examined in eutrophic sediments collected during summer stratification and fall turnover. The rate of methane formation averaged 1,130 μmol of CH
4
per liter of sediment per day during late-summer stratification versus 433 μmol of CH
4
per liter of sediment per day during the early portion of fall turnover, whereas the rate of sulfate reduction was 280 μmol of sulfate per liter of sediment per day versus 1,840 μmol of sulfate per liter of sediment per day during the same time periods, respectively. The sulfate-reducing population remained constant while the methanogenic population decreased by one to two orders of magnitude during turnover. The acetate concentration increased from 32 to 81 μmol per liter of sediment while the acetate transformation rate constant decreased from 3.22 to 0.70 per h, respectively, during stratification versus turnover. Acetate accounted for nearly 100% of total sedimentary methanogenesis during turnover versus 70% during stratification. The fraction of
14
CO
2
produced from all
14
C-labeled substrates examined was 10 to 40% higher during fall turnover than during stratification. The addition of sulfate, thiosulfate, or sulfur to stratified sediments mimicked fall turnover in that more CO
2
and CH
4
were produced. The addition of
Desulfovibrio vulgaris
to sulfate-amended sediments greatly enhanced the amount of CO
2
produced from either [
14
C]methanol or [2-
14
C]acetate, suggesting that H
2
consumption by sulfate reducers can alter methanol or acetate transformation by sedimentary methanogens. These data imply that turnover dynamically altered carbon transformation in eutrophic sediments such that sulfate reduction dominated over methanogenesis principally as a consequence of altering hydrogen metabolism.
Influence of Leaching Parameters on the Biological Removal of Uranium from Coal by a Filamentous Cyanobacterium
Authors: Lorenz Michael G. , Krumbein Wolfgang E.
Published: January 2020

Axenic cultures of the filamentous cyanobacterium LPP OL3 were incubated with samples of uraniumbearing coal from a German mining area. The influence of leaching parameters such as coal concentration (pulp density), initial biomass, particle size, temperature, and composition of the growth medium on the leaching of uranium from the ore by the cyanobacterial strain was studied. When low pulp densities were applied, the yield of biologically extracted uranium was optimal (reaching 96% at 1% [wt/vol] coal) and all released uranium was found in the culture liquid. Above 10% (wt/vol) coal in the medium, the amount of cell-bound uranium increased. Initial biomass concentration (protein content of the cultures) and particle size were not critical parameters of leaching by LPP OL3. However, temperature and composition of the growth medium profoundly influenced the leaching of uranium and growth of the cyanobacterium. The yield of leached uranium (at 10% [wt/vol] coal) could not be raised with a tank leaching apparatus. Also, coal ashes were not suitable substrates for the leaching of uranium by LPP OL3. In conclusion, the reactions of the cyanobacterium to variations in leaching parameters were different from reactions of acidic leaching organisms.
Inability of Pseudomonas stutzeri denitrification mutants with the phenotype of Pseudomonas aeruginosa to grow in nitrous oxide
Authors: Bryan B A , Jeter R M , Carlson C A
Published: January 2020

Pseudomonas aeruginosa PAO1 reduced nitrous oxide to dinitrogen but did not grow anaerobically in nitrous oxide. Two transposon insertion Nos- mutants of Pseudomonas stutzeri exhibited the P. aeruginosa phenotype. Growth yield studies demonstrated that nitrous oxide produced in vivo was productively respired, but nitrous oxide supplied exogenously was not. The defect may be in electron transport or in nitrous oxide uptake.
Plasmid profiles and transfer of plasmid-encoded antibiotic resistance in Lactobacillus plantarum
Authors: West C A , Warner P J
Published: January 2020

Plasmids were visualized in strains of Lactobacillus plantarum by use of a rapid method. Plasmids pIP501 and pAM beta 1 were transferred by conjugation from Streptococcus strains to Lactobacillus plantarum, and recipient strains were shown to act as donors in crosses to S. lactis. Attempts to transfer these plasmids between strains of L. plantarum were not successful.
Aconitase and citric acid fermentation by Aspergillus niger
Authors: Kubicek C P , Röhr M
Published: January 2020

In view of the often-cited theory that citric acid accumulation is caused by an inhibition of aconitase activity, the equilibrium of the reaction of aconitase was investigated by comparing in vivo steady-state concentrations of citrate and isocitrate in Aspergillus niger grown under various citric acid-producing conditions. With the equilibrium catalyzed by the A. niger enzyme in vitro, similar values were obtained. The validity of our in vivo measurements was verified by the addition of the aconitase inhibitor fluorocitrate, which appreciably elevated the citrate:isocitrate ratio. The results strongly argue against an inhibition of aconitase during citric acid fermentation.
Nine-year microflora study of an isolator-maintained immunodeficient child
Authors: Taylor G R , Kropp K D , Molina T C
Published: January 2020

A male child, maintained in a controlled environment, was monitored each month for bacteria, yeasts, and filamentous fungi recovered from the mouth, nasal passages, feces, and nine body surface sites. Three natural microbial categories became apparent. Incident microorganisms were recovered from within the isolator but did not establish permanent residence. Of the 53 incident types isolated, 20 were filamentous fungi and 4 were yeasts. Some genera, such as Fusobacterium, Lactobacillus, Neisseria, and Rothia, which were commonly found in the reference group, did not become permanent inhabitants. Transient microorganisms were repeatedly recovered but could not be demonstrated within the isolated environment at the end of the study. The loss of only a few of the 19 transient species could be associated with antimicrobial therapy. Permanent microorganisms consisted of Pencillium citrinum and 17 bacterial types, of which alpha-hemolytic streptococci, Staphylococcus edpidermidis subgroups II and V, Micrococcus groups 1 and 2, Clostridium bifermentans, and Propionibacterium acnes were recovered throughout the entire 9 years of the study. The number of CFUs recovered from each sample type was generally not unlike that from the reference group of healthy male adults. Also, the number of different aerobic species recovered from the feces was within the normal range of that of the reference group. In contrast, the number of different species recovered from all other samples was less than that commonly found in the reference group.
Responses of
Ruminococcus flavefaciens
, a Ruminal Cellulolytic Species, to Nutrient Starvation
Authors: Wachenheim Daniel E. , Hespell Robert B.
Published: January 2020

Ruminococcus flavefaciens
strain C94, a strictly anaerobic, cellulolytic ruminal bacterial species, was grown either in batch or continuous cultures (cellobiose limited or nitrogen limited) at various dilution rates. Washed cell suspensions were incubated anaerobically at 39�C without nutrients for various times up to 24 h. The effects of starvation on direct and viable cell counts, cell composition (DNA, RNA, protein, and carbohydrate), and endogenous production of volatile fatty acids by the cell suspensions were determined. In addition, the effect of the pH of the starvation buffer on direct and viable cell counts was determined. Survival of batch-grown cells during starvation was variable, with an average time for one-half the cells to lose viability (ST
50
) of 10.9 h. We found with continuous cultures that viable cell counts declined faster when the initial cell suspensions had been grown at faster dilution rates; this effect was more pronounced for suspensions that had been limited by cellobiose (ST
50
= 6.6 h at a dilution rate of 0.33 h
−1
) than for suspensions that had been limited by nitrogen (ST
50
= 9.5 h at a dilution rate of 0.33 h
−1
). With continuous cultures, viable cell counts in all cases declined faster than direct cell counts did. The rates of disappearance of specific cell components during starvation varied with the initial growth conditions, but could not be correlated with the loss of viability. Volatile fatty acid production by starving cells was very low, and acetate was the main product. Starved cells survived longer at pH 7.0 than they did at pH 5.5, and this effect of pH was greater for cellobiose-limited cells (mean ST
50
= 7.1 h) than for nitrogen-limited cells (mean ST
50
= 12 h). Although it has relatively low ST
50
values,
R. flavefaciens
has sufficient survival abilities to maintain reasonable numbers in domestic animals having maintenance or greater feed intake.
Fluorogenic assay for rapid detection of Escherichia coli in food
Authors: Moberg L J
Published: January 2020

An assay procedure to screen for Escherichia coli in foods by using 4-methylumbelliferyl-beta-D-glucuronide (MUG) incorporated into lauryl tryptose (LST) broth was evaluated. The beta-glucuronidase produced by E. coli cleaves the MUG substrate to yield a fluorescent end product. E. coli-negative samples can be identified by lack of fluorescence in LST-MUG within 24 h. MUG was not inhibitory to coliforms and E. coli. Over 1,400 food and dairy samples were tested to compare the standard three-tube most-probable-number procedure with the MUG-containing or non-MUG-containing LST procedure. LST-MUG testing detected a greater number of E. coli, with a lower false-positive rate (1.4%) and in a shorter time, than did the standard procedure. All false-positive results in the LST-MUG testing were attributable to beta-glucuronidase-producing staphylococci. No false-negative result was encountered. Use of MUG in LST broth obviates the EC broth step, allowing a 2.5-day procedure to a completed E. coli test versus the present 4- to 6-day standard most-probable-number method.
Physical and Chemical Correlates of Microbial Activity and Biomass in Composting Municipal Sewage Sludge
Authors: McKinley Vicky L. , Vestal J. Robie
Published: January 2020

Various physical and chemical parameters were monitored to evaluate their influence on the microbial communities present in composting municipal sewage sludge. Temperature, moisture content, depth, pH, protein content, total nitrogen, total carbon, lipid phosphate biomass, and the rates of microbial incorporation of substrates into lipids were measured at several times throughout the 17- to 19-day composting runs. Temperature was found to have the most consistent and dramatic effect on microbial activity and biomass. When temperatures exceeded 55 to 60�C, microbial activity fell dramatically, usually by more than 1 order of magnitude. Microbial activity was generally greatest in samples taken from the 35 to 50�C areas of the composting piles. Changes in the composition of the compost over time included increased pH, increased protein content, and decreased total organic content. The changes in these parameters appeared to reflect the microbial activity and biomass present. The results of this study indicate that the rate of composting may best be optimized by controlling the composting temperatures, provided that the other parameters fall within reasonable limits in the starting material.
Effects of Low-Temperature Acclimation and Oxygen Stress on Tocopheron Production in
Euglena gracilis
Z
Authors: Ruggeri Bruce A. , Gray Rodney J. H. , Watkins Thomas R. , Tomlins Richard I.
Published: January 2020

The effects of low-temperature acclimation and oxygen stress on tocopheron production were examined in the unicellular phytoflagellate
Euglena gracilis
Z. Cells were cultured photoheterotrophically at 27.5 � 1�C with 5% carbon dioxide-95% air and 740 microeinsteins m
−2
s
−1
(photosynthetically active radiation) and served as controls. Low-temperature acclimation (12.5 � 1�C) and high-oxygen stress (5% carbon dioxide-95% oxygen) were individually examined in the mass culturing of the algae. Chromatographic analyses demonstrated a six-to sevenfold enhancement of α-tocopherol production in temperature-stressed cells, along with a concomitant decline in the levels of α-tocotrienol and the absence of other tocopherol homologs. Oxygen-stressed cultures demonstrated the presence of high levels of α-tocopherylquinone; α-tocopheron and its homologs and precursors were absent or declined markedly. These findings are discussed in terms of the feasibility of microbial production of natural tocopherols. In addition, these results lend themselves to speculation regarding the biological role(s) of tocopherols as antioxidants and free radical scavengers in reducing photo-induced oxidative damage or lipid peroxidation toxicities or both in photosynthetically active
E. gracilis
Z.
Use of cloned genes of Pseudomonas TOL plasmid to effect biotransformation of benzoates to cis-dihydrodiols and catechols by Escherichia coli cells
Authors: Zeyer J , Lehrbach P R , Timmis K N
Published: January 2020

DNA fragments containing the xylD and xylL genes, which specify the broad-specificity enzymes toluate-1,2-dioxygenase and 3,5-cyclohexadiene-1,2-diol-1-carboxylic acid dehydrogenase, respectively, of TOL plasmid pWW0-161 of Pseudomonas putida have previously been cloned in the pBR322 vector plasmid (P.R. Lehrbach, J. Zeyer, W. Reinecke, H.-J. Knackmuss, and K. N. Timmis, J. Bacteriol. 158:1025-1032, 1984). In this study, Escherichia coli cells containing hybrid plasmids carrying the cloned xylD or xylDL genes quantitatively transformed 14C-ring- and 14C-carboxy-labeled benzoate to the pathway intermediates 3,5-cyclohexadiene-1,2-diol-1-carboxylic acid (cis-dihydrodiol) and catechol, respectively. Like P. putida cells, E. coli cells containing the xylD gene transformed a variety of chloro- and hydrocarbon-substituted benzoates. The toluate-1,2-dioxygenase produced in E. coli thus exhibited the broad-substrate-specificity properties of the enzyme in P. putida. Turnover rates by the enzymes in these two bacteria are compared.
Characterization of Rhizobia from Ineffective Alfalfa Nodules: Ability to Nodulate Bean Plants [
Phaseolus vulgaris
(L.) Savi.]
Authors: Eardly Bertrand D. , Hannaway David B. , Bottomley Peter J.
Published: January 2020

This study was initiated to characterize
Rhizobium
isolates obtained from root nodules of ineffectively nodulated, field-grown alfalfa (
Medicago sativa
L.) plants. The purpose was to determine if these isolates possessed characteristics which would explain either their ineffectiveness in N
2
fixation or their apparent ability to tolerate the moderately acid soil conditions from which they originated. Isolates were characterized by analysis of growth rate, 39°C tolerance, acid production on conventional media, and symbiotic performance. All isolates were ineffective in N
2
fixation on alfalfa, and they contained one or more anomalous characteristics. These included either slow growth rate, lack of 39°C tolerance, or lack of acid production on conventional media. Infectiveness tests on a broad range of legumes revealed that the isolates formed root nodules on
M. sativa, Medicago lupulina
L., and
Phaseolus vulgaris
(L.) Savi. (common bean). These results provide evidence that, in some situations, ineffective nodulation of
M. sativa
in the field may be due to the presence of promiscuous, native
Rhizobium
species.
Optimizing aerobic conversion of glycerol to 3-hydroxypropionaldehyde
Authors: Slininger P J , Bothast R J
Published: January 2020

When cells of Klebsiella pneumoniae NRRL B-199 (ATCC 8724) were grown aerobically on a rich glycerol medium and then suspended in buffer supplemented with semicarbazide and glycerol, aerobic conversion of glycerol to 3-hydroxypropionaldehyde (3-HPA) ensued. Depending on conditions, 0.38 to 0.67 g of 3-HPA were formed per gram of glycerol consumed. This means that up to 83.8% of the carbon invested as glycerol could potentially be recovered as the target product, 3-HPA. Production of 3-HPA was sensitive to the age of cells harvested for resuspension and was nonexistent if cells were cultivated on glucose instead of glycerol as the sole carbon source. Compared with 24- and 72-h cells, 48-h cells produced 3-HPA at the highest rate and with the greatest yield. The cell biomass concentration present during the fermentation was never particularly critical to the 3-HPA yield, but initial fermentation rates and 3-HPA accumulation displayed a linear dependence on biomass concentration that faded when biomass exceeded 3 g/liter. Fermentation performance was a function of temperature, and an optimum initial specific 3-HPA productivity occurred at 32 degrees C, although the overall 3-HPA yield increased continuously within the 25 to 37 degrees C range studied. The pH optimum based on fermentation rate was different from that based on overall yield; 8 versus 7, respectively. Initial glycerol concentrations in the 20 to 50 g/liter range optimized initial 3-HPA productivity and yield.
Concentration of viruses from water by using cellulose filters modified by in situ precipitation of ferric and aluminum hydroxides
Authors: Farrah S R , Preston D R
Published: January 2020

Untreated cellulose filters adsorbed only small amounts of poliovirus 1, echovirus 5, coxsackievirus B5, or bacteriophage MS2 that were added to tap water or to solutions of imidazole-glycine buffer at pH 5 to 7. Modification of filters by in situ flocculation of ferric and aluminum hydroxides greatly increased the ability of the filters to adsorb viruses. Viruses adsorbed to the modified filters could be recovered by treating the filters with 3% beef extract (pH 9.5). Greater than 60% of the enteroviruses and greater than 55% of the MS2 added to tap water or buffer could be recovered in the beef extract eluate.
Gamma processing of Arabic bread for immune system-compromised cancer patients
Authors: Grecz N , Brannon R , Jaw R , Al-Harithy R , Hahn E W
Published: January 2020

Arabic bread prepared from local Saudi flour contained a total of up to 10(4) organisms per g. Most of these were bacterial spores that survived the baking process (1.3 X 10(2) to 3.5 X 10(3] and a small number of yeasts and molds (10 to 40 cells per g). The organisms in Arabic bread appear to be harmless to healthy individuals. However, for immune system-compromised cancer patients and bone marrow transplant recipients, it is prudent to irradiate the bread to reduce microbial contamination. The decimal reduction doses (10% survival) for the most radiation-resistant organisms (spore formers) in bread were 0.11 to 0.15 Mrad. Accordingly, 0.6 Mrad was sufficient to reduce the number of spores in Arabic bread by a factor of 10,000, i.e., to less than 1/g. This treatment constitutes radiation pasteurization (radicidation), and to this extent, provides a margin of microbiological safety. Sensory evaluation by the nine-point hedonic scale showed no detectable loss of organoleptic quality of bread up to 0.6 Mrad, while irradiation to 2.5 Mrad induced unacceptable organoleptic changes.
Uptake and clearance of Vibrio vulnificus from Gulf coast oysters (Crassostrea virginica)
Authors: Kelly M T , Dinuzzo A
Published: January 2020

Oysters collected in late winter, when they were free of Vibrio vulnificus, were exposed in the organism in the laboratory. The oysters effectively concentrated the bacteria from seawater, but when the inoculum was removed, the bacteria were rapidly cleared from the oyster tissues. These results suggest that V. vulnificus may be found in oysters as a result of filtration of the bacteria from seawater rather than active multiplication of the bacteria in the oysters.
Selective Medium for Recovering Specific Populations of Rhizobia Introduced into Tropical Soils
Authors: Habte Mitiku
Published: January 2020

Experiments were designed to evaluate the usefulness of antifungal agents and streptomycin for recovering low densities of rhizobia inoculated into tropical soils. The results showed that yeast-mannitol agar (pH 6.0) containing 500 μg of streptomycin, 400 μg of cycloheximide and 50 μg of benomyl or chlorothalonil per ml was the best selective medium.
Effect of Nickelous and Other Metal Ions on the Inhibition of Rumen Bacterial Metabolism by 3-(3′-Isocyanocyclopent-2-Enylidene)Propionic Acid and Related Isocyanides
Authors: Brewer D. , Calder F. W. , Jones G. A. , Tanguay D. , Taylor A.
Published: January 2020

3-(3′-Isocyanocyclopent-2-enylidene)propionic acid at a concentration of 2 to 5 μg ml
−1
inhibited cellulose digestion by a mixed culture of rumen microorganisms and in other experiments inhibited the degradation of timothy hay (
Phleum pratense
) in a digestibility test. At isocyanide concentrations of 12 μg ml
−1
the fermentation activity of rumen fluid, measured by its dehydrogenase activity, was inhibited but not abolished. All of these isocyanide effects were reversed by the incorporation of nickelous ion into the solutions of the systems under study. The activity of 1 mol of isocyanide is reversed by about 1 mol of Ni
2+
and, in the case of the cellulose digestion test, by about 1 mol of Co
2+
. Of some 15 other ions tested only Pd
2+
and possibly chromium reversed the effect of the isocyanide.
Activity of an Attached and Free-Living
Vibrio
sp. as Measured by Thymidine Incorporation,
p
-Iodonitrotetrazolium Reduction, and ATP/DNA Ratios
Authors: Jeffrey Wade H. , Paul John H.
Published: January 2020

Three independent techniques, [
3
H]thymidine incorporation, the reduction rate of
p
-iodonitrotetrazolium violet (INT) to INT formazan normalized to DNA, and the ratio of ATP to DNA, were adapted to measure the activity of attached and unattached estuarine bacteria. In experiments employing the estuarine isolate
Vibrio proteolytica
, nutrient concentrations were manipulated by varying the concentration of peptone-yeast extract. In the presence of exogenous nutrients, the activity of free-living cells was greater than that of attached cells as measured by [
3
H]thymidine incorporation and ATP/DNA ratios. In the absence of peptone-yeast extract, however, the activity of attached cells surpassed that of free-living cells as determined by [
3
H]thymidine incorporation and INT formazan normalized to DNA. Of the three techniques, [
3
H]thymidine incorporation was deemed most sensitive for detecting changes in activity resulting from slight differences in nutrient concentration. By this technique, attached cells were much less sensitive to changing nutrient concentrations than were free-living cells. Below a threshold concentration, attached cell activity remained constant, while the activity of unattached cells decreased as a function of decreasing nutrient concentration. The results suggest that loss of cell surface area available for substrate uptake due to attachment may be an important factor in determining the relative activities of attached and free-living cells.
Simple method for determination of patulin production by Penicillium griseofulvum Dierckx
Authors: Torres M , Sanchis V , Riba M , Canela R
Published: January 2020

Patulin production by Penicillium griseofulvum was monitored with Sep-Pak cartridges and high-pressure liquid chromatography. Determination and quantification of this metabolite proved to be very simple, and our method saved time and a large amount of organic solvents.
Isolation of a rat intestinal Clostridium strain producing 5 alpha- and 5 beta-bile salt 3 alpha-sulfatase activity
Authors: Robben J , Parmentier G , Eyssen H
Published: January 2020

An unnamed sporeforming microorganism, termed Clostridium sp. strain S2, possessing bile salt sulfatase activity was isolated from rat intestinal microflora. The microorganism was a strictly anaerobic, nonmotile, gram-negative, asaccharolytic, sporeforming rod requiring CO2, vitamin K, and taurine; the guanine-plus-cytosine content of the DNA was 40.8 mol% (Tm), and the strain was tentatively classified as an atypical Clostridium species. Sulfatase activity was specific for 3 alpha-sulfate esters of 5 alpha- and 5 beta-bile salts, leaving the 3 beta-, 7 alpha-, and 12 alpha-sulfates unchanged. Strain S2 also deconjugated tauro- and glyco-conjugated bile salts and partially reduced into the corresponding 6 alpha-hydroxy bile salts. By these reactions, alpha-muricholate and beta-muricholate were more than 80% converted into hyocholate and omega-muricholate, respectively. In addition, strain S2 produced 12 alpha-hydroxysteroid dehydrogenase converting deoxycholate into 3 alpha-hydroxy-12-oxo-5 beta-cholanoate. When strain S2 was associated with gnotobiotic rats, the fecal bile salts were more than 90% desulfated and the fecal excretion of allochenodeoxycholate was five times lower than in control rats.
Production of toxin by Clostridium botulinum type A strains cured by plasmids
Authors: Weickert M J , Chambliss G H , Sugiyama H
Published: January 2020

Twelve strains of Clostridium botulinum type A and seven strains of Clostridium sporogenes were screened for plasmids by agarose gel electrophoresis of cleared lysates of cells from 5 ml of mid-log-phase culture. Nine type A strains had one or more plasmids of 4.3, 6.8, or 36 megadaltons (MDa); several strains showed a large plasmid of 61 MDa, but it was not consistently recovered. Four C. sporogenes strains had one or more plasmids of 4.3, 5.6 or 36 MDa. Isolates obtained from cultures of plasmid-containing C. botulinum type A strains grown in ionic detergent broth and from spontaneously arising variants were screened both for toxin production and for plasmid content. Toxigenicity of C. botulinum could not be correlated with the presence of any one plasmid.