bluuebunny/crossref_metadata_2025 huggingface dataset:
Bacterial Growth Kinetics on Diphenylmethane and Naphthalene-Heptamethylnonane Mixtures
Authors: Wodzinski Richard S. , Larocca David
Published: January 2020

Experiments were carried out to determine if diphenylmethane is utilized by a species of
Pseudomonas (Hydrogenomonas)
in the dissolved state regardless of the physical state (liquid or solid) of the undissolved diphenylmethane suspended in the medium. Bacterial growth rates in the presence of various amounts of solid or liquid diphenylmethane indicate that liquid diphenylmethane is utilized at the aqueous-diphenylmethane interface but that solid diphenylmethane is not. A
Pseudomonas
sp. that was isolated on naphthalene (solid), but could not utilize heptamethylnonane, was grown in the presence of various amounts of a naphthalene-heptamethylnonane mixture (liquid). The growth rates indicate that the bacterium could utilize naphthalene at the aqueous-hydrocarbon interface, which is not the case in the absence of the heptamethylnonane.
Sensitivity of Various Bacteria, Including Actinomycetes, and Fungi to Cadmium and the Influence of pH on Sensitivity
Authors: Babich H. , Stotzky G.
Published: January 2020

A variety of microorganisms, including gram-negative and gram-positive eubacteria, actinomycetes, yeasts, and filamentous fungi, were tested for their sensitivity to cadmium (Cd). In general, the actinomycetes were more tolerant to Cd than were the eubacteria; gram-negative eubacteria were more tolerant to Cd than were gram-positive eubacteria. The period of exponential growth of the eubacteria and actinomycetes was extended in the presence of Cd. Wide extremes in sensitivity to Cd were noted among the fungi; there was no correlation between the class of fungus and tolerance to Cd. Fungal sporulation was more sensitive to Cd than was mycelial growth, as spore formation was inhibited at Cd concentrations that were noninhibitory to mycelial proliferation. The toxicity of Cd to the eubacteria, actinomycetes, and fungi appeared to be pH dependent, as toxicity was generally potentiated at pH 8 or 9.
Tentative Identification of Methanogenic Bacteria by Fluorescence Microscopy
Authors: Mink Ronald W. , Dugan Patrick R.
Published: January 2020

Methanogenic bacteria, which are presently identified on the basis of cell morphology and substrate conversion to CH
4
, can be differentiated from nonmethanogens and identified in pure or mixed culture on the basis of their autofluorescence under ultraviolet illumination.
Characterization of a Cytoplasmic Reserve Glucan from
Ruminococcus albus
Authors: Cheng K.-J. , Brown R. G. , Costerton J. W.
Published: January 2020

Ruminococcus albus
, an anaerobic bacterium that digests cellulose in the rumen of cattle, produces intracellular polysaccharide granules varying from 0.05 to 0.31 μm in diameter when grown in batch culture. This polysaccharide material was purified and found to contain
d
-glucose as the only reducing sugar. The polyglucose polymer was slightly opalescent in aqueous solution and formed a strong reddish purple iodine complex with a maximum absorbance at 550 nm. Its infrared spectrum had characteristic absorption bands at 8.70, 9.25, and 9.75 μm and was identical with that of the amylopectin-glycogen type of
Megasphaera elsdenii
and that of the glycogen of enteric bacteria and beef liver. It reacted strongly with concanavalin A. Methylation analysis showed that the glucan contained 2,3,4,6-tetra-
O
-MeG-2,3,6-Tri-
O
-MeG-2,3-Di-
O
-MeG in a ratio of 8:84:8. Characterization of the products obtained by treatment with isoamylase indicates that the glucan of
R. albus
is of the glycogen type.
Induction and Catabolite Repression of Glucosidases in
Pseudomonas maltophilia
Authors: Boethling R. S.
Published: January 2020

Intracellular α-and β-glucosidases were induced in cell suspensions of
Pseu-domonas maltophilia
by maltose or cellobiose, and the synthesis of these enzymes was sensitive to apparent catabolite repression by α-ketoglutarate.
Isolation of Atypical
Candida albicans
from the North Sea
Authors: Crow S. A. , Bowman P. I. , Ahearn D. G.
Published: January 2020

Isolates of
Candida albicans
with sparse filamentation and weak fermentation were isolated from the surface microlayer of the North Sea, but not from subsurface waters. Such atypical isolates may be misidentified by using normal taxonomic procedures.
Produciton and biological activity of patulin and citrinin from Penicillium expansum
Authors: Ciegler A , Vesonder R F , Jackson L K
Published: January 2020

Penicillium expansum isolated from meat and apples produced both patulin and citrinin. Toxin identity was confirmed by spectroscopic and physical methods. The mean lethal dose in chicken embryos was determined for toxins administered both singly and in various ratios. Data from simultaneous administration of mycotoxin combinations plotted as isobolograms showed and additive effect. Both toxins were teratogenic in chicken embryos.
Recovery of Clostridia on Catalase-Treated Plating Media
Authors: Harmon Stanley M. , Kautter Donald A.
Published: January 2020

Four plating media commonly used for culturing clostridia were tested for their ability to support growth of several
Clostridium
species after storage of the plates for 1 to 10 days at 4 and 25°C with and without subsequent addition of catalase. Liver-veal (LV) agar and brain heart infusion (BHI) agar rapidly became incapable of supporting growth after storage without added catalase, whereas Shahidi Ferguson perfringens agar base and Brewer anaerobic agar were less affected. Plate counts of vegetative cells of nine of the less fastidious
Clostridium
species on untreated LV and BHI agars, stored for 3 days at 4°C, were 60 to 90% lower than counts on catalase-treated media. Counts on Shahidi Ferguson perfringens agar base were only 1 to 24% lower on untreated medium with the same species. Addition of 500 U of purified beef liver catalase to the surface of the 3-day-old agars before inoculation resulted in substantial restoration of the ability of the media to support colony formation from vegetative cells except with the most strictly anaerobic species (nonproteolytic
C. botulinum
types B, E, and F, and
C. novyii
types A and B). A similar response was obtained with spores of the less fastidious species on catalase-treated media. Our results suggest that inhibition of most
Clostridium
species on LV and BHI agars may be due to accumulation of peroxide during preparation, storage, and incubation of the media, and also suggest that the presence of glucose in these media is a major factor contributing to their inability to support growth. It is believed that the addition of exogenous catalase prevents the accumulation of peroxide(s), thus allowing colony formation from vegetative cells of the clostridia under what would otherwise be unsuitable cultural conditions.
Oxidation of linear terpenes and squalene variants by Arthrobacter sp
Authors: Yamada Y , Kusuhara N , Okada H
Published: January 2020

Cells of Arthrobacter sp. that had been isolated from soil were used to study oxidation of some linear terpenes and squalene variants. The cells oxidized geraniol, nerol, and farnesol to the corresponding aldehydes, with partial conversion of the geometrical isomerism of the alpha,beta-double bond. The squalene variant, squalene-2,3-oxide, was cleaved to 9,10-epoxygeranylacetone and geranylacetone. Squalene-2,3-22,23-dioxide was cleaved to 9,10-epoxygeranylacetone. These products were optically active, and their stereochemistry and optical purity were determined.
Succession of Streptococcus bovis strains with differing bacteriophage sensitivities in the rumens of two fistulated sheep
Authors: Iverson W G , Millis N F
Published: January 2020

The bacteriophage sensitivity of the Streptococcus bovis population resident in the rumens of two fistulated sheep was monitored for 112 days. During this time, three changes in the bacteriophage sensitivity of S. bovis occurred in the absence of detectable bacteriophages. Identical changes in bacteriophage sensitivity occurred simultaneously in both animals and, except for the relatively short periods of changeover in phage sensitivity, the S. bovis population in the rumens of the two sheep was homogeneous with respect to phage sensitivity.
Nitrogen Fixation (Acetylene Reduction) in a Salt Marsh Amended with Sewage Sludge and Organic Carbon and Nitrogen Compounds
Authors: Hanson Roger B.
Published: January 2020

Seasonal distribution of nitrogen fixation by
Spartina alterniflora
epiphytes and in surface and soil samples was investigated in a Georgia salt marsh which was amended with sewage sludge or with glucose and/or ammonium nitrate. There was no significant difference between the rates of fixation in the unamended and sewage sludge plots. Additional perturbation experiments suggested that nitrogen addition indirectly stimulates nitrogen fixation by enhancing
Spartina
production and root exudation. Glucose additions, on the other hand, suppressed nitrogen fixation on a long-term basis. It is suggested that the microbial population in the soil out-competed the plants for the available nitrogen and in turn suppressed plant production and possibly root exudation. A comparison of nitrogen fixation in clipped and unclipped
Spartina
plots substantiated the suggestion that root exudation probably supports nitrogen fixation. Fixation in the clipped plots was significantly lower (
P
< 0.05) than the rates in the unclipped plots.
Microbiological characteristics of Pacific shrimp (Pandalus jordani)
Authors: Lee J S , Pfeifer D K
Published: January 2020

Microorganisms associated with Pacific shrimp (Pandalus jordani) were isolated and identified. Those on the iced raw shrimp, which yielded an average count of 1.6 x 10(6), were predominantly Moraxella, Pseudomonas, Acinetobacter, Arthrobacter, and Flavobacterium-Cytophaga spp. The blanching and peeling reduced the microbial level to 3.3 x 10(4) and also selectively eliminated Moraxella spp. The microbial flora changed after each processing sequence, and the heat sensitivity and growth characteristics of the representative microbial groups suggested that the presence of Arthrobacter and Acinetobacter spp. in peeled shrimp may indicate inadequate cleaning of raw shrimp or a shorter blanching time. The presence of Moraxella and Flavobacterium-Cytophaga spp. would indicate the degree of secondary contamination, and the presence of Pseudomonas spp. would indicate the shelf-age of the processed shrimp.
Energetics of Streptococcal Growth Inhibition by Hydrostatic Pressure
Authors: Matsumura Philip , Marquis Robert E.
Published: January 2020

Growth of
Streptococcus faecalis
in complex media with various fuel sources appeared to be limited by the rate of supply of adenosine-5′ -triphosphate (ATP) at 1 atm and also under 408 atm of hydrostatic pressure. Growth under pressure was energetically inefficient, as indicated by an average cell yield for exponentially growing cultures of only 10.7 g (dry weight) per mol of ATP produced compared with a 1-atm value of 15.6. Use of ATP for pressure-volume work or for turnover of protein, peptidoglycan, or stable ribonucleic acid (RNA) did not appear to be significant causes of growth inefficiency under pressure. In addition, there did not seem to be an increased ATP requirement for ion uptake because cells growing at 408 atm had significantly lower internal K
+
levels than did those growing at 1 atm. Pressure did stimulate the membrane adenosine triphosphatase (ATPase) or
S. faecalis
at ATP concentrations greater than 0.5 mM. Intracellular ATP levels were found to vary during the culture cycle from about 2.5 μmol/ml of cytoplasmic water for lag-phase or stationary-phase cells to maxima for exponentially growing cells of about 7.5 μmol/ml at 1 atm and 5.5 μmol/ml at 408 atm.
N,N′
-dicyclohexylcarbodiimide at a 10 μM concentration improved growth efficiency under pressure, as did Mg
2+
or Ca
2+
ions at 50 mM concentration. These agents also enhanced ATP pooling, and it seemed that at least part of the growth inefficiency under pressure was due to increased ATPase activity. In all, it appeared that
S. faecalis
growing under pressure has somewhat reduced ATP supply but significantly increased demand and that the inhibitory effects of pressure can be interpreted largely in terms of ATP supply and demand.
Numerically Dominant Denitrifying Bacteria from World Soils
Authors: Gamble Thomas N. , Betlach Michael R. , Tiedje James M.
Published: January 2020

Nineteen soils, three freshwater lake sediments, and oxidized poultry manure were examined to determine the dominant denitrifier populations. The samples, most shown or expected to support active denitrification, were from eight countries and included rice paddy, temperate agricultural, rain forest, organic, and waste-treated soils. Over 1,500 organisms that could grow anaerobically on nitrate agar were isolated. After purification, 146 denitrifiers were obtained, as verified by production of N
2
from NO
3
-
. These isolates were characterized by 52 properties appropriate for the
Pseudomonas-Alcaligenes
group. Numerical taxonomic procedures were used to group the isolates and compare them with nine known denitrifier species. The major group isolated was representative of
Pseudonomas fluorescens
biotype II. The second most prevalent group was representative of
Alcaligenes
. Other
Pseudomonas
species as well as members of the genus
Flavobacterium
, the latter previously not known to denitrify, also were identified. One-third of the isolates could not utilize glucose or other carbohydrates as sole carbon sources. Significantly, none of the numerically dominant denitrifiers we isolated resembled the most studied species:
Pseudomonas denitrificans, Pseudomonas perfectomarinus
, and
Paracoccus denitrificans
. Denitrification appears to be a property of a very diverse group of gram-negative, motile bacteria, as shown by the large number (22.6%) of ungrouped organisms. The diversity of denitrifiers from a given sample was usually high, with at least two groups present. Denitrifiers, nitrite accumulators, and organisms capable of anaerobic growth were present in the ratio of 0.20±0.23:0.81±0.23:1. There were few correlations between their numbers and the sample characteristics measured. However, the temperatures at which isolates could grow were significantly related to the temperatures of the environments from which they were isolated. Regression analysis revealed few relationships between physical parameters and bacterial types, save for the anaerobe numbers, in which 94% of the variance could be accounted for.
Use of enzyme-labeled antibodies to detect Salmonella in foods
Authors: Krysinski E P , Heimsch R C
Published: January 2020

An indirect enzyme-labeled antibody technique (ELAT), in which Salmonella typhimurium was used as a model, was developed as a method to detect Salmonella in food samples. A cellulose-acetate membrane filter, the matrix for detection, was placed on a membrane-filter base and overlaid with a multiwelled lucite template. Mixed broth enrichment cultures were dispensed in the template wells, and cells were spotted onto the membrane via suction. After fixation, the membranes were immersed in rabbit anti-S. typhimurium flagella antibody, washed, immersed in goat anti-rabbit antibody conjugated to peroxidase, and washed. Exposure of membranes to the substrates 3,3'-diaminobenzidine or benzidine resulted in development of brown or blue macroscopic reaction products, respectively, on spots containing S. typhimurium. ELAT results agreed with those of enrichment serology and cultural procedures on three food products containing known levels of S. typhimurium. Because of the magnification effect of the enzyme-substrate reaction, fewer cells were needed for detection than with enrichment serology, thereby reducing the total analysis time. The ability to test 14 or more samples simultaneously on a 47-mm membrane filter would facilitate screening large number of samples. Pending the development of a pure H antisera pool for the common Salmonella serotypes free from O antibodies, the ELAT demonstrated potential as a Salmonella detection methodology.
Antigenicity of bdellovibrios
Authors: Kramer T T , Westergaard J M
Published: January 2020

Antigenic relationships between 12 locally isolated bdellovibrios and 3 established reference strains (109D, 6-5-S, and UKi2) were investigated. Antigenicity of the strains was examined by use of the micro-complement fixation test, the serum and complement bactericidal test, and the immunodiffusion test. Antisera were prepared against one of the local strains (MS7) and against one of the established reference strains (UKi2). The complement fixation titers suggest a close relationship among all strains. Immunodiffusion tests produced lines of identity between the homologous strain MS7 and all other strains. It is suggested on the basis of these results that bdellovibrio may possess a common antigen.
Linkage of Mercury, Cadmium, and Arsenate and Drug Resistance in Clinical Isolates of
Pseudomonas aeruginosa
Authors: Nakahara Hideomi , Ishikawa Tomoaki , Sarai Yasunaga , Kondo Isamu , Kozukue Hiroyuki , Silver Simon
Published: January 2020

Of the 787 isolates, 99.8% were metal resistant, with most (99.5%) showing multiple resistance. Fifty-three percent of the isolates were both metal and drug resistant, whereas only 19% were metal resistant and drug sensitive.
Rapid tube test for detecting fungal cellulase production
Authors: Smith R E
Published: January 2020

A simple procedure is described for the detection of cellulolytic activity in fungi, utilizing dyed cellulose powder as substrate. The powder, incorporated into a basal medium containing agar, mineral salts, and growth factors, is layered onto basal medium minus cellulose. Cellulase production results in release of the dye which diffuses into the lower layer.
Small pet aquarium frogs as a source of Salmonella
Authors: Bartlett K H , Trust T J , Lior H
Published: January 2020

Salmonellae were isolated from 21% of the samples of freshwater aquarium frogs tested and from 25% of the samples of aquarium water containing these frogs. The salmonellae were Salmonella arizonae, S. bovis-morbificans, S. hadar, S. saint-paul, S. typhimurium, and S. worthington. These isolations were made over a period of 9 months and from three different cities. This association of salmonellae with frogs may contribute to cases of human salmonellosis since other aquarium species have already been shown to contribute to such cases.
Indonesian Tapé Ketan Fermentation
Authors: Cronk T. C. , Steinkraus K. H. , Hackler L. R. , Mattick L. R.
Published: January 2020

Indonesian tapé ketan is a fermentation in which a mold,
Amylomyces rouxii
Calmette (
Chlamydomucor oryzae
Went and Prinsen Geerligs), in combination with one or more yeasts such as
Endomycopsis burtonii
converts steamed rice to a sweet-sour, slightly alcoholic paste. A study was made to determine the biochemical changes that occur in the substrate during fermentation. It was found that the product was ready for consumption after fermentation at 30°C for 36 to 48 h.
A. rouxii
used about 30% of the total rice solids, resulting in a crude protein of 12% in 96 h, whereas the combination of the mold with
E. burtonii
reduced total solids by 50% in 192 h, causing crude protein to increase to 16.5%. Soluble solids increased from 5 to about 67% in 36 h and decreased to 12% at 192 h with
A. rouxii
alone, whereas soluble solids fell to about 8% at 192 h in the fermentation with both the mold and the yeast. The mold, by itself, reduced the starch content of the rice from 78 to 10% in 48 h and to less than 2% in 144 h. The mold plus yeast reduced the starch content to about 18% in 48 h; however the “starch” content did not fall below 6% even at 192 h, presumably because the yeast was producing glycogen, which was determined along with the residual starch. With both the mold and the mold plus yeast fermentations, reducing sugars increased from less than 1% to approximately 5% in 24 h and reached maximum concentration, 16 to 17%, between 36 and 48 h.
A. rouxii
by itself produced a maximum of about 5.6% (vol/vol) ethanol at 96 h. The highest concentration of ethanol (8%, vol/vol) was produced by the mold plus
E. burtonii
at 144 h. The mold by itself reduced the starting pH from 6.3 to about 4.0 in 48 h. The combination of the mold and yeast reduced the pH to 4.1 in 144 h. The mold increased total acidity to approximately 6.2 meq of H
+
per 100 ml, and the combination of the mold and yeast increased the total acidity to 7.8 meq of H
+
per 100 ml in 192 h. At 48 h there was practically no difference in the volatile acidity (0.20) for the combined fermentation compared with 0.26 meq of H
+
per 100 ml for the mold fermentation. The mold and at least one species of yeast were required to develop the rich aroma and flavor of typical Indonesian tapé.
Growth of heat-injured Vibrio parahaemolyticus in media supplemented with various cations
Authors: Heinis J J , Beuchat L R , Jones W K
Published: January 2020

Mid- to late logarithmic growth phase cells of Vibrio parahaemolyticus grown in tryptic soy broth (TSB) containing 0.5, 3.0, and 7.5% NaCl were heated for 8 min at 45 degrees C in 0.1 M phosphate buffer (pH 7.2) containing 3% NaCl. Colony formation on thiosulfate-citrate-bile salts-sucrose agar (TCBS) containing 2% NaCl was greatest for unheated cells that had been grown in 7.5% NaCl-TSB; cells grown in 0.5% NaCl-TSB formed a greater number of colonies on 1.0% NaCl-TCBS. Thermal injury was evident in heated cells, regardless of the NaCl concentration in TSB growth medium. The effects of Mg2+, K+, and Li+ added as chlorides to 0.5% NaCl-TSB on the growth of nonheated and heated V. parahaemolyticus were studied. Lower levels of Mg2+ and slightly higher levels of K+ were required to replace Na+ in TSB inoculated with thermally injured cells that had been originally grown in 3.0 and 7.5% NaCl-TSB. LiCl had an inhibitory effect on both nonheated and heated cells when present in the recovery medium (0.5% NaCl-TSB) at concentrations as low as 0.5%. Increased numbers of colonies were formed by heated cells plated in MgCl2-supplemented TCBS, regardless of the NaCl concentration in the original growth medium. Potassium had little, if any, effect on colony formation by nonheated V. parahaemolyticus recovered on TCBS and may have had a detrimental effect on heat-injured cells.
Laminarinase (beta-glucanase) activity in Bacteroides from the human colon
Authors: Salyers A A , Palmer J K , Wilkins T D
Published: January 2020

Laminarin, a beta(1 leads to 3)-glucan similar to those found in plant cell walls, is fermented by some species of anaerobic bacteria from the human colon. Laminarinase (EC 3.2.1.6) and beta-glucosidase (EC 3.2.1.21) activities were determined in strains representing Bacteroides thetaiotaomicron, Bacteroides distasonis, and an unnamed deoxyribonucleic acid homology group of Bacteroides fragilis. In all three species, laminarinase activity was inducible by laminarin and was predominantly cell bound. The products of laminarinase activity varied with each species. In the case of B. thetaiotaomicron, the major product of laminarin hydrolysis was glucose (70 to 90%), and there were small amounts of laminaribiose (G2) and oligomers of glucose as high as G4. In the case of group '0061-1,' glucose (40 to 50%) and oligomers of glucose as high as G6 were found. The laminarinase of B. distasonis differed from the laminarinases of the other two species in that it mainly produced oligomers of glucose (G2-G5). beta-Glucosidase activity was also found in all three species. beta-Glucosidase was induced by glucose-containing disaccharides as well as by laminarin. The beta-glucosidases of the three Bacteroides species differed with respect to level of activity, induction pattern, and sensitivity to inhibition by D-glucono-1,5-lactone.
Minimal requirements in defined media for improved growth of some radio-resistant pink tetracocci
Authors: Shapiro A , DiLello D , Loudis M C , Keller D E , Hutner S H
Published: January 2020

Defined media permitting extensive growth of representative pink radio-resistant tetracocci (Micrococcus radiodurans, Micrococcus roseus, and Micrococcus radiophilus) and two controls (an ultraviolet-sensitive mutant of M. radiodurans and Micrococcus luteus) are described. Availability of Fe (especially Fe3+) proved essential for good growth, as evidenced by (i) favorable effects of hydroxamic acids, e.g., salicylhydroxamic acid, and (ii) the growth promotion by hemin when joined with elevated concentrations of Fe. Cobalamin (B12) and methionine were interchangeable as an absolute requirement for methionine not affected by B12. M. luteus required neither. Pink radio-resistant micrococci may form a coherent group. Some divergences among them might be attributable to the method for isolating them, which for ordinary bacteria would be mutagenic to the point of total lethality. The ecology of these tetracocci vis-à-vis other pink-red radio-resistant organisms is discussed in relation to a question: can these bacteria be isolated without dependence on radiation as the cardinal selective factor?
Evaluation of a new presumptive medium for group D streptococci
Authors: Abshire R L
Published: January 2020

A new medium designated as D streptococcus-enterococcus broth was formulated and evaluated for the enrichment and isolation of strains of serological group D streptococci. This medium was made by modifying Todd-Hewitt broth. Most-probable-number multiple-tube and membrane filter techniques were employed to estimate the numbers of enterococci in known cultures, wastewater, and other samples. Preliminary most-probable-number counts with this medium were as much as 3 logs higher than those counts obtained from four other media with which it was compared. The methodology for using this medium to estimate the numbers of group D streptococci in water is discussed.
Use of nuclepore filters for counting bacteria by fluorescence microscopy
Authors: Hobbie J E , Daley R J , Jasper S
Published: January 2020

Polycarbonate Nuclepore filters are better than cellulose filters for the direct counting of bacteria because they have uniform pore size and a flat surface that retains all of the bacteria on top of the filter. Although cellulose filters also retain all of the bacteria, many are trapped inside the filter where they cannot be counted. Before use, the Nuclepore filters must be dyed with irgalan black to eliminate autofluorescence. Direct counts of bacteria in lake and ocean waters are twice as high with Nuclepore filters as with cellulose filters.
Two-temperature membrane filter method for enumerating fecal coliform bacteria from chlorinated effluents
Authors: Green B L , Clausen E M , Litsky W
Published: January 2020

Reports indicate that the standard membrane filter (MF) technique for recovery of fecal coliform bacteria from chlorinated sewage effluents is less effective than the multiple-tube (or most-probable-number [MPN]) procedure. A modified MF method was developed that requires a preincubation period of 5 h at 35 degrees C followed by 18+/-1 h at 44.5 degrees C. This procedure was evaluated by using both laboratory- and plant-chlorinated primary and secondary effluents. Results obtained by the modified MF method compared favorably with those of the MPN technique for the enumeration of fecal coliforms from chlorinated effluent. Agreement between these two methods was greatest with samples from secondary treatment plants. The average recovery of fecal coliforms by the standard MF procedure was only 14% that of the MPN method, whereas with the modified technique recovery was increased to 68% of the MPN counts. Enhanced recovery resulting from a simple modification in the incubation schedule makes the MF method a valuable adjunct for enumerating fecal coliforms from chlorinated effluents.
Identification of intermediates formed during the degradation of hexachlorocyclohexanes by Clostridium sphenoides
Authors: Heritage A D , Rae I C
Published: January 2020

Washed cell suspensions of Clostridium sphenoides degraded the alpha-isomer of 1,2,3,4,5,6-hexachlorocyclohexane via delta-3,4,5,6-tetrachloro-1-cyclohexene and the gamma-isomer via gamma-3,4,5,6-tetrachloro-1-cyclohexene. Both intermediates were further metabolized to unknown substances. The tetrachlorocyclohexene intermediates were identified by gas chromatography and mass spectrometry.
Rapid induction of alpha-amylase by nongrowing mycelia of Aspergillus oryzae
Authors: Yabuki M , Ono N , Hoshino K , Fukui S
Published: January 2020

A rapid induction system for synthesis of alpha-amylase by the funga Aspergillus oryzae M-13 was established. The mycelia were prepared from 20-h cultures grown on a peptone-glycerol medium and starved for 5 h; maltose was the optimum inducer tested. During h 1 of induction, formation of both intra- and extracellular alpha-amylases occurred at an almost identical rate (70 to 80 microgram/g of cells-h) without a detectable lag period. After a 1-h induction period, a remarkable increase in the extracellular concentration of the enzyme occurred, and a maximum rate (330 microgram/g of cells-h) was reached after 1.5 h of induction. During h 2 of induction, no significant change in mycelial weight was observed. Purified samples of intra- and extracellular enzymes formed in the induction system showed identical properties as examined by behavior in diethylaminoethyl-cellulose column chromatography, gel filtration, discontinuous gel electrophoresis, electrofocusing, optimal conditions for the reaction, heat stability, and molecular weight.
Effect of dredge spoil deposition on fecal coliform counts in sediments at a disposal site
Authors: Babinchak J A , Graikoski J T , Dudley S , Nitkowski M F
Published: January 2020

The most-probable-number of fecal coliforms in sediments was monitored at the New London dump site in Long Island Sound during the deposition of dredge spoil from the Thames River. Although the geometric mean for fecal coliforms at five stations in the river was 14,000/100 ml before dredging commenced, the deposition of this material did not increase the incidence of fecal coliforms at 17 spoil stations and 13 control stations in the disposal and surrounding areas. Fecal coliforms appear to occur only in the surface sediment material and are diluted by the subsurface material during the dredging operation. Fecal coliform analyses of bottom waters during high and low tides indicated that the flow of water from the Thames River played a major role in determining the most-probable-number of fecal coliforms in the sediments at the disposal site.
Stereospecific sulfur oxidation of 7-methylthioxanthone-2-carboxylic acid by Calonectria decora
Authors: Lanzilotta R P , Bradley D G , Maddox M L
Published: January 2020

Thin-layer chromatography was used to determine the ability of three microorganisms capable of sulfur oxygenation, including Aspergillus niger, Streptomyces armentosus subsp. armentosus, and Calonectria decora, to oxidize 7-methylthioxanthone-2-carboxylic acid to the corresponding sulfoxide in growing cultures. In addition, optical rotary dispersion, circular dichroism, and nuclear magnetic resonance analysis in the presence of chiral shift reagent were used variously to access reaction stereoselectivity, absolute configuration, and optical purity of isolated products. The data indicated that C. decora produced the sulfoxide in high yield (69%) and optical purity (97%), most probably in the S-configuration.
Comparison of the surface structure, metal binding, and fecal coliform recoveries of nine membrane filters
Authors: Tobin R S , Dutka B J
Published: January 2020

A comparative study was made of nine commonly used membrane filters from five manufacturers, all recommended for enumeration of coliform bacteria. Bacterial recoveries and flow rates were examined from three types of water and were found to correlate with the surface pore structure determined by scanning electron microscopy. The sorption of metals was also determined. The results of these studies indicate that the five best membranes for fecal coliform recovery could be placed in two groups: Millipore HC and Gelman, followed by Johns-Manville SG and AG and Sartorius 13806.
Aflatoxin localization by the enzyme-linked immunocytochemical technique
Authors: Lawellin D W , Grant D W , Joyce B K
Published: January 2020

Visualization of aflatoxin deposits within Aspergillus parasiticus ATCC 15517 hyphae was made possible by using an enzyme-linked immunocytochemical technique.
Enzyme-linked immunosorbent analysis for aflatoxin B1
Authors: Lawellin D W , Grant D W , Joyce B K
Published: January 2020

An enzyme-linked immunosorbent analysis (ELISA) permitted the detection of less than 10 pg of aflatoxin B1 per ml. The antitoxin was most specific for aflatoxins B1 and B2alpha, and least specific for aflatoxin G1.
Proteolytic mutants obtained from Clostridium botulinum type E
Authors: Nakane A , Iida H
Published: January 2020

Proteolytic mutants were isolated from toxigenic strains of Clostridium botulinum type E after several transfers. When these cultures were plated on blood agar, almost all of the colonies obtained were proteolytic, and there were fewer toxigenic colonies than nontoxigenic colonies. The proteolytic mutants and nonproteolytic original strains were different in their biological properties.
Rapid spectrophotometric differentiation between glutathione-dependent and glutathione-independent gentisate and homogentisate pathways
Authors: Crawford R L , Frick T D
Published: January 2020

A total of four pathways are known for the catabolism by microorganisms of gentisate (2,5-dihydroxybenzoate) and homogentisate (2,5-dihydroxyphenylacetate). Both of these dihydric phenols can be degraded by either a glutathione-dependent or a glutathione-independent reaction sequence. We found that it is not always possible to unequivocally assign glutathione dependence or independence to a particular catabolic sequence by using the well-established spectrophotometric assays at 330 nm (gentisate pathway) or 320 nm (homogentisate pathway). This paper reports a modification of the classical spectrophotometric assays that allowed an unequivocal differentiation between glutathion-dependent and glutathione-independent pathways, even when crude cell extracts contained significant quantities of cell-derived, reduced glutathione. This was accomplished by performing assays in the presence of an approximately 10(-3) M solution of the sulfhydryl-binding agent N-ethylmaleimide.
Nose, throat, and fecal flora of beagle dogs housed in "locked" or "open" environments
Authors: Balish E , Cleven D , Brown J , Yale C E
Published: January 2020

The microbial flora of the nose, throat, and feces of male beagle dogs housed in a "locked environment" (i.e. confined to germfree-style isolators and supplied with sterile food, air, and water) or an open environment were assessed between 26 and 30 months into the study. Forty-five genera and 170 different species or types of microorganisms were cultured from the nose, throat, and feces of the beagles. Clostridia, eubacteria, corynebacteria, bacteroides, lactobacilli, and anaerobic, gram-positive cocci accounted for most of the microbial diversity in the flora. Some of the facultative anaerobes, especially streptococci and lactobacilli (in feces), occurred in numbers that were comparable to the most numerous anaerobic species. Confinement to the locked environment resulted in an increased diversity of microorganisms in the flora, but the total microbial counts did not increase to any great extent. Even with the increased diversity of bacteria in the flora of confined dogs, some bacteria seemed to favor certain areas of the gastrointestinal tract over others. The increased diversity of bacteria observed in these confined dogs may pose some infectious disease problems for other mammals (including humans) that may be confined to a locked, ultra-clean environment for a prolonged period of time.
Isolation, extraction, and measurement of acetylcholine from Lactobacillus plantarum
Authors: Stanaszek P M , Snell J F , O'Neill J J
Published: January 2020

The isolation, extraction, and spectrophotometric determination of acetylcholine from Lactobacillus plantarum ATCC 10241 is described. Acetylcholine was extracted with a mixture of sodium tetraphenylboron-butylethylketone-acetonitrile and was measured enzymatically at 340 nm.
Removal of Escherichia coli in wastewater by activated sludge
Authors: van der Drift C , van Seggelen E , Stumm C , Hol W , Tuinte J
Published: January 2020

Removal of bacteria from wastewater treated with activated sludge was studied by the use of a streptomycin-resistant Escherichia coli strain. The removal appeared to be a biphasic process. A rapid sorption of bacteria to the sludge flocs took place in the first hour after seeding mixed liquor with E. coli. Thereafter, slower elimination of E. coli was observed. The latter process was due to predation on E. coli by ciliated protozoa. This was shown by: (i) appearance of fluorescent food vacuoles of ciliates when fluorescent E. coli cells were added to mixed liquor; (ii) inhibition of predation either in the presence of cycloheximide or under anaerobic conditions; and (iii) absence of predation in bulking and washed sludge.
Mutation of an inosine-producing strain of Bacillus subtilis to DL-methionine sulfoxide resistance for guanosine production
Authors: Matsui H , Sato K , Enei H , Hirose Y
Published: January 2020

An inosine-producing strain of Bacillus subtilis was mutated to resistance against the antagonist of glutamine, DL-methionine sulfoxide. Among the mutants derived, guanosine producers were observed frequently. The best strain, 14119, produced 9.6 g of guanosine per liter at a weight yield of 12% from consumed sugar. Inosine production decreased concomitantly. When resistance was increased further by exposure to higher doses of DL-methionine sulfoxide, another strain, AG169, was obtained that did not excrete inosine but produced increased amounts of xanthosine. In these strains, the specific activity of 5'-nucleotidase was lower and that of inosine 5'-monophosphate (IMP) dehydrogenase was higher than the parent strain. It is speculated that the metabolic flow from IMP to xanthosine 5'-monophosphate proceeds more smoothly than that from IMP to inosine and yields more xanthosine and guanosine.
Structure, Growth, and Decomposition of Laminated Algal-Bacterial Mats in Alkaline Hot Springs
Authors: Doemel W. N. , Brock Thomas D.
Published: January 2020

Laminated mats of unique character in siliceous alkaline hot springs of Yellowstone Park are formed predominantly by two organisms, a unicellular blue-green alga,
Synechococcus lividus
, and a filamentous, gliding, photosynthetic bacterium,
Chloroflexus aurantiacus
. The mats can be divided approximately into two major zones: an upper, aerobic zone in which sufficient light penetrates for net photosynthesis, and a lower, anaerobic zone, where photosynthesis does not occur and decomposition is the dominant process. Growth of the mat was followed by marking the mat surface with silicon carbide particles. The motile
Chloroflexus
migrates vertically at night, due to positive aerotaxis, responding to reduced O
2
levels induced by dark respiration. The growth rates of mats were estimated at about 50 μm/day. Observations of a single mat at Octopus Spring showed that despite the rapid growth rate, the thickness of the mat remained essentially constant, and silicon carbide layers placed on the surface gradually moved to the bottom of the mat, showing that decomposition was taking place. There was a rapid initial rate of decomposition, with an apparent half-time of about 1 month, followed by a slower period of decomposition with a half-time of about 12 months. Within a year, complete decomposition of a mat of about 2-cm thickness can occur. Also, the region in which decomposition occurs is strictly anaerobic, showing that complete decomposition of organic matter from these organisms can occur in the absence of O
2
.
Uptake and Incorporation of Glucose and Mannose by Whole Cells of
Bacteroides thetaiotaomicron
Authors: Hylemon P. B. , Young J. L. , Roadcap R. F. , Phibbs P. V.
Published: January 2020

Glucose uptake by whole-cell suspensions of the obligate anaerobe
Bacteroides thetaiotaomicron
was two- to fourfold higher under aerobic conditions than during incubation under atmospheres of N
2
or H
2
gas. The O
2
-stimulated uptake activity was lost rapidly (>70% in 5 h) when cell suspensions were incubated aerobically, but this loss was prevented by the addition of crude catalase. Catalase had no apparent effect on cell viability during these incubations. Glucose uptake activity was strongly inhibited by a 10-fold excess of mannose or galactose but not by methyl-α-
d
-glucoside, fructose, or lactose. Both glucose and mannose were rapidly incorporated into polyglucose after uptake. The O
2
-stimulated glucose uptake was not inhibited by cyanide, azide, 2,4-dinitrophenol, or 2-
N
-heptyl-4-hydroxyquinoline-
N
-oxide. However,
p
-chloromercuribenzoate, menadione, and sodium fluoride inhibited uptake by 88, 67, and 55%, respectively. All attempts to detect phosphoenolpyruvate-phosphotransferase activity for glucose, methyl-α-
d
-glucoside, and 2-deoxyglucose were negative. The bacteria contained hexokinase activity and a complete glycolytic Embden-Meyerhof pathway.
High-viability lyophilized Bacille Calmette-Guérin vaccine produced by deep-culture technique
Authors: Kim T H
Published: January 2020

Evidence suggests that Bacille Calmette-Guérin (BCG) vaccine for use in cancer immunotherapy should have the following characteristics: high viability which is maintained on storage; high ratio of live to dead cells; high proportion of single cells; and low content of soluble antigen. The production of a vaccine with these characteristics was accomplished by use of a deep-culture technique. The medium was modified Proskauer and Beck medium containing Tween 80 and glucose. The mass culture was grown in a Wheaton double-side-arm bottle (6 liters of medium in an 8-liter container), aerated by means of an aquarium aerator and mixed by a magnetic stirrer. The culture was incubated 7 to 9 days at 37 degrees C, concentrated 11 to 15 times by ultrafiltration, diluted with equal parts of 25% lactose, and then lyophilized. The lyophilized ampoules, stored at -70 degrees C, were cultured at intervals ranging from 3 days to 450 days, and no loss in viability was observed. The mean number of viable BCG per ml of reconstituted vaccine was 8.75 log10. The viable count was 90% of the total bacterial count. Moreover, 85% of the cells were present as single bacilli.
Nitrification in the intertidal zone: influence of effluent type and effect of tannin on nitrifiers
Authors: Bohlool B B , Schmidt E L , Beasley C
Published: January 2020

Nitrification by intertidal sediments was measured by using a tide simulator that approximated the cycle of seawater on tidal flats. Sediments were chosen from sites affected by industrial and municipal effluents and pastoral seepage and runoff. The ability of sediments from different sites to nitrify endogenous nitrogen varied markedly. All sites exhibited an initial lag before activity commenced. The duration of this lag and the rate of nitrate production were different at each site. The sediments were also capable of oxidizing NH3-N supplied to them in seawater. This "nitrification potential" was highest at sites receiving nitrogenous effluents (slaughterhouse and sewage), but was also substantial in sediments affected by bark extract effluent and pasture runoff. The lowest potential and the longest lag were exhibited by sediments in an apple cannery effluent area. Enrichment cultures of nitrifying microorganisms were obtained from all sites using NH4+ as a source of energy, but enrichments for nitrite oxidizers were unsuccessful. Concentrated pine bark tannins, similar in origin to those in effluents at the well-nitrifying chipmill site, were tested for toxicity to pure cultures of nitrifying bacteria. Two Nitrobacter strains and one Nitrosomonas strain were unaffected by tannins even at 5 mg/ml. A Nitrosolobus and a Nitrosospira strain were inhibited partially at 5 mg/ml and only slightly or not at all at 1 mg/ml.
Effect of an activated carbon filter on the microbial quality of water
Authors: Fiore J V , Babineau R A
Published: January 2020

Recently, there has been growing concern that microbial health hazards can be increased by the use of activated carbon filters in domestic water systems. The present study was undertaken to investigate the effect of carbon filters on the microbial content of water. Results indicated that the microbial content of filtered and unfiltered water increased to about the same level on overnight standing and, in both cases, was reduced by flushing the next day. In addition, the use of activated carbon for the filtration of contaminated well water over a period of 11 weeks had no effect on the total or coliform count. Under use conditions, activated carbon filters were found to have no significant effect on the number of bacteria present in the water.
Penicillic acid production in submerged culture
Authors: Lindenfelser L A , Ciegler A
Published: January 2020

Twenty known penicillic acid (PA)-producing Aspergillus and Penicillium cultures were grown under various conditions in shaken flasks to determine the highest yielding strains and their requirements for maximum toxin production. Abilities of the cultures to utilize eight different carbon sources in Raulin-Thom medium for mycotoxin synthesis were determined at four different incubation temperatures: 15, 20, 25, and 28 degrees C. Of the 20 cultures, P. cyclopium NRRL 1888 was superior, yielding up to 4 mg of PG per ml, with mannitol as the carbon source and 25 degrees C as the incubation temperature. Fifteen of the cultures elaborated lesser amounts of PA, whereas four strains yielded none under the test conditions. Whey from the manufacture of cottage cheese by the cultured process was also a satisfactory medium for PA production. In whey medium, yields up to 3 mg/ml were obtained with P. cyclopium NRRL 1888.
Colonization of the Botanical Environment by
Klebsiella
Isolates of Pathogenic Origin
Authors: Knittel Martin D. , Seidler Ramon J. , Eby Chris , Cabe Linda M.
Published: January 2020

Growth, survival, and pathogenicity of
Klebsiella
growing in and on environmental foci were examined. Total coliforms present in raw wastes from pulp mills were in excess of 10
5
/ml, and 60 to 80% were
Klebsiella
. Fecal coliform counts ranged from 10
1
to 10
5
/ml.
Klebsiella
isolates from industrial effluents and a variety of human and bovine mastitis origins multiplied in pulp waste and commonly exceeded 10
6
cells per ml. Pathogenic isolates also multiplied in dilute aqueous extracts of sawdust to comparable levels.
Klebsiella
strains from vegetable surfaces and human infections grew rapidly on the surfaces of potatoes and lettuce and exceeded 10
3
organisms per g of surface peel and leaf after a 24h incubation at room temperature. After 7 weeks on potatoes stored at 5°C, some 10 to 30% of the day 1
Klebsiella
counts were recoverable. Three
Klebsiella
isolates of pathogenic origin were passed 45 times through sterile pulp effluent (270 generations), and mean lethal dose levels in mice were periodically monitored. In two instances, a significant decrease in virulence was noted after 15 to 26 passes (90 to 156 generations). The third culture, of bovine mastitis origin, retained its original mean lethal dose value. Botanical milieu provided suitable habitats for the multiplication and colonization of
Klebsiella
isolates of disease origins in the same manner as indigenous isolates. Aquatic environments polluted with botanical material served as potential reservoirs for perpetuating the growth and spread of opportunistic
Klebsiella
pathogens that may ultimately colonize animals, humans, and aquatic organisms.