bluuebunny/crossref_metadata_2025 huggingface dataset:
Adsorption of bacteria to roots as related to host specificity in the Rhizobium-clover symbiosis
Authors: Dazzo F B , Napoli C A , Hubbell D H
Published: January 2020

Quantitative microscope techniques were utilized to examine the adsorption of rhizobial cells to clover root hairs. Adsorption of cells of noninfective strains of Rhizobium trifolii or infective R. meliloti strains to clover root hairs was four to five times less than that of the infective R. trifolii strains. Attachment of the rod-shaped bacteria to clover root cells occurred in a polar, end-on fashion. Viable or heat-killed R. trifolii cells precoated with a clover lectin having 2-deoxyglucose specificity had increased adsorption to clover roots. Adsorption of bacteria to roots was not increased if the clover lectin was inactivated by heat or 2-deoxyglucose treatment prior to incubation with R. trifolii. Adsorption of R. trifolii to clover root hairs was inhibited by 2-deoxyglucose (30 mM) but not by 2-deoxygalactose or alpha-D-glucose. Adsorption of R. meliloti cells to alfalfa root hairs was not affected by 2-deoxyglucose at that concentration. These results suggest that expression of host specificity in the Rhizobium-clover symbiosis involves a preferential adsorption of infective cells to clover root hairs through a 2-deoxyglucose-sensitive receptor site.
Easily Constructed Soil Percolation Apparatus
Authors: Longden A. R. , Claridge C. A.
Published: January 2020

The construction of a soil percolator from a disposable Nalgene sterilizing filter is described.
Isolation of exocellular polymer from Zoogloea strains MP6 and 106 and from activated sludge
Authors: Farrah S R , Unz R F
Published: January 2020

Exocellular polymer was isolated from zoogloeae of Zoogloea strains MP6 and 106 and from activated sludge flocs by blending samples with phosphate buffer and precipitation of solubilized polymer with cetyltrimethylammonium bromide. Samples of polymer from these sources were similar and yielded amino sugars as the principal components after acid hydrolysis.
Enzymatic hydrolysis of organophosphate insecticides, a possible pesticide disposal method
Authors: Munnecke D M
Published: January 2020

A crude cell extract from a mixed bacterial culture growing on parathion, an organophosphate insecticide, hydrolyzed parathion (21 C) at a rate of 416 nmol/min per mg of protein. This rate of enzymatic hydrolysis, when compared with chemical hydrolysis by 0.1 N sodium hydroxide at 40 C, was 2, 450 times faster. Eight of 12 commonly used organophosphate insecticides were enzymatically hydrolyzed with this enzyme preparation at rates ranging from 12 to 1,360 nmol/min per mg of protein. Seven pesticides were hydrolyzed at rates significantly higher (40 to 1,005 times faster) than chemical hydrolysis. The pH optimum for enzymatic hydrolysis of the eight pesticides ranged from 8.5 to 9.5, with less than 50% of maximal activity expressed at pH 7.0. Maximal enzyme activity occurred at 35 C. The crude extract lost its activity at the rate of only 0.75%/day when stored at 6 C. Eight organic solvents, ranging from methanol to hexane, at low concentrations stimulated enzymatic hydrolysis by 3 to 20%, whereas at higher concentrations (1,000 mg/liter) they inhibited the reaction (9 to 50%). Parathion metabolites p-nitrophenol, hydroquinone, and diethylthiophosphoric acid, at up to 100-mg/liter concentrations, did not significantly influence enzyme activity.
Utilization of Lactose, Glucose, and Galactose by a Mixed Culture of
Streptococcus thermophilus
and
Lactobacillus bulgaricus
in Milk Treated with Lactase Enzyme
Authors: O'Leary Virginia S. , Woychik John H.
Published: January 2020

The mechanism responsible for an increased rate of acid production when yogurt starter cultures are grown in milk treated with lactase enzyme was investigated by studying carbohydrate utilization and acid development by a pure culture of
Streptococcus thermophilus
and a mixed yogurt starter culture consisting of
S. thermophilus
and
Lactobacillus bulgaricus
. In milk containing glucose, galactose, and lactose, glucose and lactose (but not free galactose) were fermented. Fermentation of lactose in control milk was accompanied by the release of free galactose, with the result that carbohydrate utilization was less efficient than in treated milk. This phenomenon also occurred when lactose was fermented by
S. thermophilus
in broth culture. Carbohydrate utilization by the mixed yogurt culture was more rapid when the lactose in milk was partially prehydrolyzed. Our results suggest that the more rapid acid development that took place when a mixed yogurt starter culture was grown in milk containing prehydrolyzed lactose was the result of a more rapid and efficient utilization of carbohydrate by
S. thermophilus
when free glucose in addition to lactose was available for fermentation. The evidence presented also suggests that uptake and utilization of glucose and lactose by
S. thermophilus
are different in broth and milk cultures.
Evaluation of millipore HA and HC membrane filters for the enumeration of indicator bacteria
Authors: Lin S D
Published: January 2020

Millipore type HA and HC membrane filters were compared for recoveries of total coliforms, fecal coliforms and fecal streptococci from natural waters and wastewater effluents. Bacterial and fecal coliform recovery was better on HC than on HA filters.
Microbial Conversion of Tall Oil Sterols to C
19
Steroids
Authors: Conner Anthony H. , Nagaoka Muneo , Rowe John W. , Perlman D.
Published: January 2020

Mycobacterium
sp. NRRL B-3683 converted tall oil sterols to C
19
steroids as efficiently as it converted soybean sterols.
Heat inactivation of poliovirus in wastewater sludge
Authors: Ward R L , Ashley C S , Moseley R H
Published: January 2020

The effect of raw and anaerobically digested sludge on heat inactivation of poliovirus was investigated. Raw sludge was found to be very protective of poliovirus plaque-forming ability at all temperatures studied, but digested sludge had variable effects that were highly dependent upon the experimental conditions. In low concentrations and at relatively low inactivation temperatures, digested sludge is nearly as protective of poliovirus as raw sludge. However, at higher tempeatures and concentrations, digested sludge caused a significant acceleration of poliovirus inactivation. The difference between the protective capability of raw and digested sludge is not due to loss of protective material, because this component is present in the solids of digested sludge as well as in those of raw sludge. Instead, the difference is due to a virucidal agent acquired during digestion. Addition of this agent to the solids of either raw or digested sludge reverses the protective potential of these solids during heat treatment of poliovirus.
Subsampling technique for measuring growth of bacterial cultures under high hydrostatic pressure
Authors: Taylor C D , Jannasch H W
Published: January 2020

A method is presented for measuring growth of bacteria under high hydrostatic pressure in subsamples taken without pressure change in the incubation vessel. Subsamples may be withdrawn rapidly (5 s) and are not subjected to shear forces. Vice versa, nutrient media, labeled substrates, etc., may be introduced into the culture while under pressure. Chemical fixation of subsamples for electron microscopy or adenosine 5'-triphosphate determinations under pressure is also possible without affecting the growing culture. Data are given of growth experiments demonstrating the feasibility of the method. Problems of oxygen depletion are discussed.
Factors affecting production of mold mycelium and protein in synthetic media
Authors: Graham D C , Steinkraus K H , Hackler L R
Published: January 2020

The effects of certain cultural conditions on the yield of dry mycelium, protein, and total amino acid content of Rhizopus oligosporus Saito (NRRL 2710), Rhizopus rhizopodiformis (Cohn apud Lichtheim) Zopf (NRRL 6246), and Absidia corymbifera (Cohn) Sacc. et Trotter (NRRL 6247) were studied. The yield of mycelium was found to significantly increase as the spore inoculum was increased from 187,500 to 2,250,000 spores. But the total amino acids (grams/liter) did not change significantly, whereas the percentage of crude protein decreased. An inoculum containing approximately 750,000 spores/ml was used in all of the other experiments. Mycelial production was highest at 37 degrees C for all three molds. However, the best temperature for percentage of crude protein and total amino acids varied with the organism. The mycelial yield and total crude protein of R. oligosporus showed some significant changes as the C/N ratio was increased in 3% glucose medium. In a synthetic medium having a 15:1 C/N ratio, the strains of R. oligosporus, R. rhizopodiformis, and A. corymbifera had better yields from falactose than glucose, not only in dry mycelium but also in total crude protein (grams/liter) and total amino acids (grams/liter). R. oligosporus grew very well on several ammonium salts. but the maximum yield of dry mycelium, total crude protein (grams/liter), and total amino acids (grams/liter) occurred with ammonium sulfate. The optimum pH for both Rhizopus species was 4.0, although R. oligosporus grew equally well at pH 3.0 and slightly less at pH 5.0. The highest yield of mycelium for A. corymbifera was obtained in a medium with an initial pH of 8.0. It was calculated that a fermenter chanrged with an adequate medium and 1,000 lb (about 450 kg) of R. oligosporus or A. corymbifera cells could produce 88 or 90 lb of protein (on a dry-weight basis) per h if the product was removed continuously.
Protoplasts of
Cosmarium
as a Potential Protein Source
Authors: Berliner Martha D. , Wenc Katherine A.
Published: January 2020

Cosmarium turpinii
, a fast-growing desmid alga, transforms into protoplasts in 4 h when incubated in a mineral medium + 0.4 M mannitol + 0.5% Cellulysin.
Comparison of Various Brands of Membrane Filters for Their Ability to Recover Fungi from Water
Authors: Qureshi A. A. , Dutka B. J.
Published: January 2020

Five brands of ethylene oxide- and autoclave-sterilized membrane filters were examined for their ability to recover fungi from natural waters. Results showed that the recovery on Gelman membranes was consistently higher than on the other brands tested.
Analysis of the Exudate Produced by
Streptococcus mutans
SL-1 Colonies on Sucrose-Containing Agar Media
Authors: Donkersloot J. A. , de Leon H. A. , Chassy B. M. , Krichevsky M. I.
Published: January 2020

The watery exudate produced by
Streptococcus mutans
SL-1 colonies on sucrose-containing agar media was found to contain about 7% (wt/vol) of a water-soluble, branched dextran, 4% sucrose, and smaller (less than 1%) amounts of fructose, Folin-phenol-positive material, and lactic acid.
Presence of bifidobacteria in the rumen of calves fed different rations
Authors: Trovatelli L D , Matteuzzi D
Published: January 2020

A study was made on the numbers and species of bifidobacteria present in the rumen of calves fed high-roughage and high-concentrate diets. With the roughage ration the bifidobacteria were not detectable in a 10(-3) dilution, whereas with the concentrate ration their number was high, usually in the order of 10(8) to 10(9)/ml of rumen fluid. The species most represented, identified by means of deoxyribonucleic acid-deoxyribonucleic acid hybridization tests, included Bifidobacterium ruminale, Bifidobacterium globosum, and an apparently new species.
Regulation of Lactose Fermentation in Group N Streptococci
Authors: Thomas Terence D.
Published: January 2020

Group N streptococci, which have the lactose phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) and phospho-β-
d
-galactosidase (β-Pgal), grew rapidly on lactose and converted more than 90% of the sugar to
l
-lactate. In contrast,
Streptococcus lactis
7962, which does not have a β-Pgal, grew slowly on lactose and converted only 15% of the sugar to
l
-lactate. With glucose and galactose, this strain had growth rates and fermentation patterns similar to those of other
S. lactis
strains, suggesting that the rapid and homolactic fermentation of lactose that is characteristic of group N streptococci is dependent upon a functional PEP-dependent PTS and the presence of β-Pgal. Seventeen strains of group N streptococci were examined for the activator specificities of pyruvate kinase and lactate dehydrogenase. The properties of each enzyme from all the strains, including
S. lactis
7962, were similar. Pyruvate kinase had a broad activator specificity, whereas activation of lactate dehydrogenase was specific for ketohexose diphosphate. All intermediates of lactose metabolism from the hexose phosphates to the triose phosphates activated pyruvate kinase. No activation was obtained with adenosine 5′-monophosphate. K
+
and Mg
2+
were required for pyruvate kinase activity but could be replaced by NH
4
+
and Mn
2+
, respectively. Lactate dehydrogenase was activated equally by fructose-1,6-diphosphate and tagatose-1,6-diphosphate, the activation characteristics being pH dependent. The roles of pyruvate kinase and lactate dehydrogenase in the regulation of lactose fermentation by group N streptococci are discussed.
Preparation of Agar-Agar from the Red Seaweed
Pterocladia capillacea
off the Coast of Alexandria, Egypt
Authors: Rao A. V. , Bekheet Inaam A.
Published: January 2020

The effect of different treatments on the quality of agar produced from
Pterocladia
has been studied, and the conditions for the production of material of good quality have been standardized. In the modified process, sun-bleached seaweed was washed well in water, soaked for 24 h, and then ground to a pulp and rinsed again in water. The pulp was then extracted with water (weed-to-water ratio, 1:30) under pressure for 2 h after adjusting the pH to 6 by the addition of acetic acid. The agar gel, after freeze thawing, was bleached with NaClO before drying in a current of hot air. Pretreatment of the seaweed with alkali at 80°C for 2 h prior to extraction was found to improve the quality of agar to a very great extent.
Regulation of nitrogen fixation in Rhizobium sp
Authors: Tubb R S
Published: January 2020

Regulation of nitrogen fixation by ammonium and glutamate was examined in Rhizobium sp. 32H1 growing in defined liquid media. Whereas nitrogenase synthesis in Klebsiella pneunoniae is normally completely repressed during growth on NH4+, nitrogenase activity was detected in cultures of Rhizobium sp. grown with excess NH4+. However, an "ammonium effect" on activity was invariably observed in cultures grown on NH4+ as sole nitrogen source; the nitrogenase activity was, depending on conditions, 14 to 36% of that of comparable glutamate-grown cultures. Glutamate inhibited utilization of exogenous NH4+ and, in one of two procedures described, glutamate partially alleviated the ammonium effect on nitrogenase activity. NH4+, apparently produced from N2, was excreted into the culture medium when growth was initiated on glutamate, but not when NH4+ was thesole source of fixed nitrogen for growth. These findings are discussed in relation to nitrogen fixation by Rhizobium bacteroids.
Studies on glucose isomerase from a Streptomyces species
Authors: Chou C C , Ladisch M R , Tsao G T
Published: January 2020

Production and properties of glucose isomerase from a Co2+-sensitive Streptomyces species were studied. After 4 days of shaking cultivation at 30 degrees C and 200 rpm, a maximum of 1.1 enzyme units per ml of broth was obtained. Cell-free glucose isomerase, obtained from mycelia heat-treated in the presence of 0.5 mM Co2+, showed a 3.5-fold increase in specific activity over enzyme obtained from untreated mycelia. The optimum pH and temperature for the glucose isomerase were 7 to 8 and 80 degrees C, respectively. The Michaelis constant for fructose was 0.40 M. Mg2+ was found to enhance the glucose isomerase activity, whereas the effect of Co2+ on enzyme activity depended on the manner in which the enzyme was prepared. This glucose isomerase was quite heat stable, with a half-life of 120 h at 70 degrees C.
Determinants of the developing oral flora in normal newborns
Authors: Long S S , Swenson R M
Published: January 2020

The ability of Streptococcus species to selectively adhere to the oral epithelial cells of newborns was studied in vitro. On day 1 of life, mucosal cells from normal infants demonstrated selective attraction for the natural distribution of streptococci that would soon colonize these surfaces. Streptococcus salivarius and Streptococcus mitis adhered well in vitro to scraped cells from cheek and tongue surfaces. Streptococcus mutans, on the other hand, exhibited feeble or no adherence to cheek or tongue cells. Adherence of Escherichia coli to oral epithelial cells was also studied. The ability of strains of E. coli to adhere to cheek and tongue cells correlated solely with the presence of cell surface substances, probably pili. These observations, made on infants at the critical moment of their developing flora, strengthen the hypothesis that the ability of bacteria to adhere to surfaces is an important determinant of their ecological place in the oral microflora.
Mercurial toxicity in yeast: evidence for catabolic pathway inhibition
Authors: Brunker R L
Published: January 2020

Evidence that the mechanism of mercurial toxicity is a blockage of catabolic metabolism is presented. Yeast cells (Saccharomyces cerevisiae) were found to cease respiratory activities within 1.5 min of contrast time with culture mercurials (as HgCl2). This cessation was followed by the rapid depletion of endogenous adenosine 5'-triphosphate (ATP) and a concomitant increase in phosphorylated hexoses. Levels of ATP in the culture medium remained essentially unchanged during this interval suggesting that the structural integrity of the membrane was not affected. Medium potassium concentrations did not increase until after endogenous ATP levels had begun to fall, suggesting that the loss of cellular potassium was the result of the inability of membrane ATPases to function because of the unavailability of sufficient substrate ATP to maintain this gradient.
Virus movement in soil columns flooded with secondary sewage effluent
Authors: Lance J C , Gerba C P , Melnick J L
Published: January 2020

Secondary sewage effluent containing about 3 X 10(4) plaque-forming units of polio virus type 1 (LSc) per ml was passed through columns 250 cm in length packed with calcareous sand from an area in the Salt River bed used for ground-water recharge of secondary sewage effluent. Viruses were not detected in 1-ml samples extracted from the columns below the 160-cm level. However, viruses were detected in 5 of 43 100-ml samples of the column drainage water. Most of the viruses were adsorbed in the top 5 cm of soil. Virus removal was not affected by the infiltration rate, which varied between 15 and 55 cm/day. Flooding a column continuosly for 27 days with the sewage water virus mixture did not saturate the top few centimeters of soil with viruses and did not seem to affect virus movement. Flooding with deionized water caused virus desorption from the soil and increased their movement through the columns. Adding CaCl2 to the deionized water prevented most of the virus desorption. Adding a pulse of deionized water followed by sewage water started a virus front moving through the columns, but the viruses were readsorbed and none was detected in outflow samples. Drying the soil for 1 day between applying the virus and flooding with deionized water greatly reduced desorption, and drying for 5 days prevented desorption. Large reductions (99.99% or more) of virus would be expected after passage of secondary sewage effluent through 250 cm of the calcareous sand similar to that used in our laboratory columns unless heavy rains fell within 1 day after the application of sewage stopped. Such virus movement could be minimized by the proper management of flooding and drying cycles.
Fecal indicator bacteria persistence under natural conditions in an ice-covered river
Authors: Davenport C V , Sparrow E B , Gordon R C
Published: January 2020

Total coliform (TC), fecal coliform (FC), and fecal streptococcus (FS) survival characteristics, under natural conditions at 0 degrees C in an ice-covered river, were examined during February and March 1975. The membrane filter (MF) technique was used throughout the study, and the multiple-tube (MPN) method was used in parallel on three preselected days for comparative recovery of these bacteria. Survival was studied at seven sample stations downstream from all domestic pollution sources in a 317-km reach of the river having 7.1 days mean flow time (range of 6.0 to 9.1 days). The mean indicator bacteria densities decreased continuously at successive stations in this reach and, after adjustment for dilution, the most rapid die-off was found to occur during the first 1.9 days, followed by a slower decrease. After 7.1 days, the relative survival was TC less than FC less than FS, with 8.4%, 15.7%, and 32.8% of the initial populations remaining viable, respectively. These rates are higher than previously reported and suggest that the highest survival rates for these bacteria in receiving streams can be expected at 0 degree C under ice cover. Additionally, the FC-FS ratio was greater than 5 at all stations, indicating that this ratio may be useable for determining the source of fecal pollution in receiving streams for greater than 7 days flow time at low water temperatures. The MPN and MF methods gave comparable results for the TC and FS at all seven sample stations, with both the direct and verified MF counts within the 95% confidence limits of the respective MPNs in most samples, but generally lower than the MPN index. Although FC recovery on membrane filters was comparable results at stations near the pollution source. However, the results became more comparable with increasing flow time. The results of this study indicate that heat shock is a major factor in suppression of the FC counts on the membrane filters at 44.5 degree C. Heat shock may be minimized by extended incubation at 35 degrees C before exposure to the higher temperature.
Membrane filter method for recovery of fecal coliforms in chlorinated sewage effluents
Authors: Lin S D
Published: January 2020

The standard one-step M-FC broth-membrane-filter procedure for recovery of fecal coliforms from chlorinated sewage effluents is much less effective than the multiple-tube (most-probable-number) technique. A two-step membrane-filter method, using a pre-enrichment technique with phenol red lactose broth and incubation at 35 degrees C for 4 h, followether 18+/-2 h, enhanced fecal coliform recovery from chlorinated effluents. The results of 126 comparisons using chlorinated effluents from five wastewater plants showed that fecal coliform recovery by using the two-step membrane-filter method is comparable to that using the multiple-tube procedure.
Natural occurrence of Fusarium toxins in feedstuff
Authors: Mirocha C J , Pathre S V , Schauerhamer B , Christensen C M
Published: January 2020

The mycotoxins diacetoxyscirpenol, deoxynivalenol, and zearalenone, produced by Fusarium roseum, were found naturally occuring in mixed feed samples. In all cases analyzed, deoxynivalenol occurred together with zearalenone. The natural occurrence of zearalenone in sesame seed is reported for the first time. Strains of F. roseum isolated in various parts of the world form feed implicated in animal mycotoxicosis produced monoacetoxyscirpenol, diacetoxyscirpenol, deoxynivalenol, and zearalenone.
Morphological and physiological characteristics of Gemmiger formicilis isolated from chicken ceca
Authors: Salanitro J P , Muirhead P A , Goodman J R
Published: January 2020

Morphological and physiological studies were made on chicken cecal isolates of the strictly anaerobic bacterial species Gemmiger formicilis. Structural features (phase-contrast and electron microscopy) of these microorganisms indicate they (i) are highly pleomorphic, (ii) possess a trilaminar cell wall like gram-negative bacteria, (iii) exhibit an unusual growth process characterized by polar swelling (resembling budding bacteria), and (iv) grow into elongated cells when exposed to a subinhibitory concentration of penicillin. The morphological data presented suggest that this species has a rod-shaped structure. These bacteria ferment a variety of sugars to produce formic, butyric, and lactic acids. There appear to be two groups of Gemmiger, one producing primarily lactate and the other producing formate as major fermentation metabolites. Growth of six strains in a basal medium, consisting of Trypticase, minerals, carbohydrate, Na2CO3 buffer, and cysteine as reducing agent, was stimulated by rumen fluid and yeast extract. Volatile fatty acids partially replaced the requirement for rumen fluid with some strains. Single deletions of vitamins (from a defined vitamin mixture) indicated that pantothenate, riboflavin, and thiamine were highly stimulatory to growth of the organism in a medium containing rumen fluid and Trypticase as source of vitamins. Other vitamin requirements were not studied.
Organic Flocculation: an Efficient Second-Step Concentration Method for the Detection of Viruses in Tap Water
Authors: Katzenelson Eliyahu , Fattal Badri , Hostovesky Tova
Published: January 2020

A method is described for second-step concentration of viruses from water. This method, combined with an adsorption-elution method, yields a mean recovery of about 75%
Linear Transformation of Standard Curves for Yeast Turbidity
Authors: Bestic Philip B. , Arnold Wilfred N.
Published: January 2020

Within a very serviceable range, light extinction plotted against yeast concentration follows a rectangular hyperbola. A linear transformation, which facilitates the determination of yeast concentration, is described.
Effect of alfalfa fiber substrate on culture counts of rumen bacteria
Authors: Chung K T , Hungate R E
Published: January 2020

A medium has been developed using alfalfa fiber as the sole substrate. It gave high culture counts (3 X 10(9) to 8 X 10(9)/ml) of rumen bacteria. When this medium was combined with the medium 98-5 of Bryant and Robinson, modified to contain 33% rumen fluid instead of 40% clarified rumen fluid, a higher count was obtained than with either medium alone.
Alcohol Dehydrogenase Activities of Wine Yeasts in Relation to Higher Alcohol Formation
Authors: Singh Rajendra , Kunkee Ralph E.
Published: January 2020

Alcohol dehydrogenase activities were examined in cell-free extracts of 10 representative wine yeast strains having various productivities of higher alcohols (fusel oil). The amount of fusel alcohols (
n
-propanol, isobutanol, active pentanol, and isopentanol) produced by the different yeasts and the specific alcohol dehydrogenase activities with the corresponding alcohols as substrates were found to be significantly related. No such relationship was found for ethanol. The amounts of higher alcohols formed during vinification could be predicted from the specific activities of the alcohol dehydrogenases with high accuracy. The results suggest a close relationship between the control of the activities of alcohol dehydrogenase and the formation of fusel oil alcohols. Also, new procedures for the prediction of higher alcohol formation during alcoholic beverage fermentation are suggested.
Inactivation of Clostridium botulinum toxin by ruminal microbes from cattle and sheep
Authors: Allison M J , Maloy S E , Matson R R
Published: January 2020

Toxin from Clostridium botulinum type C was rapidly inactivated during incubation in vitro with ruminal contents from either a cow or a sheep. Fractions of ruminal contents from which cells had been removed by high-speed centrifugation did not inactivate toxin. Inactivation was associated with fractions containing bacteria, whereas fractions containing protozoa and relatively few bacteria were much less active. This activity may help explain the relatively greater tolerance by ruminants to oral doses of botulinum toxin than to toxin administered by other routes. The results are also pertinent to assays for botulinum toxin from gastrointestinal samples obtained postmortem.
Basal medium for the selective enumeration of rumen bacteria utilizing specific energy sources
Authors: Dehority B A , Grubb J A
Published: January 2020

A 40% rumen fluid basal medium has been developed that without added substrate will support growth of about 10% or less of the total colony count obtained with 40% rumen fluid-glucose-cellobiose-starch-agar medium (RGCSA). The basal medium is prepared by anaerobic incubation of all ingredients in RGCSA medium except the carbohydrates, Na2CO3, and cysteine for 7 days at 38 degrees C. After incubation, substrate(s), Na2CO3 and cysteine are added and the medium is tubed and sterilized as in normal medium preparation. When xylose was included with glucose, cellobiose, and starch as added carbohydrates in the incubated medium, colony counts were comparable to those obtained with RGCSA medium. The addition of specific carbohydrates or other substrates as energy sources to the basal medium suggested that the percentage of the bacterial population capable of utilizing these energy sources was influenced by the ration of the animal; however, considerable animal variation and day-to-day variation in a given animal was observed. Comparison of the population in animals fed either orchardgrass hay or 60% corn-40% orchardgrass (60-40) indicated little or no difference for the percentage of bacteria utilizing glucose, pectin, xylan, or mannitol. Increases in the percentages of xylose-, cellobiose-, Glycerol-, and lactate-utilizing bacteria occurred with the orchardgrass hay ration, whereas the percentage of starch-digesting bacteria was increased significantly (P less than 0.01) in the animals fed the 60-40 ration. A limited number of bacterial strains were isolated from the basal medium without added substrate, most of which were atypical with respect to the predominant rumen bacteria. Growth of these strains, even in complex media, was very slow and limited. Based on these data with isolated strains and colony counts obtained in roll tube medium containing only minerals, resazurin, agar, Na2CO3, and cysteine, the selective medium overestimated the percentage of bacteria able to use a specific energy source. This overestimate was 6 to 7% of the total culturable count.
Timed-release capsule method for coliform enumeration
Authors: Lanz W W , Hartman P A
Published: January 2020

Wax-coated capsules containing selective ingredients (brilliant green and oxgall) were added at the time of inoculation of most-probable-number media (modified lactose broths). The inhibitory ingredients gradually diffused from the capsules into the nonselective media, imparting selectivity to the media. Concentrations of brilliant green did not reach inhibitory levels until 2 or more h had elapsed, which permitted repair of some injured cells. Resuscitation of heat-injured Escherichia coli B cells occurred in the capsule-containing media, but not in conventional brilliant green bile 2% broth or violet red bile agar. No statistically significant differences were noted between coliform counts obtained on two groups of water samples by using the capsule, most-probable-number, membrane filtration, and pour plate methods. The capsule method could be used, however, as a combined presumptive and confirmed test for the examination of water. Improvements are needed to adapt the capsule method to the analysis of some categories of food.
Microbiological Hydroxylation of Carbons 18 and 19 in 9-Oxo-13 (
cis
and
trans
)-Prostenoic Acids
Authors: Lanzilotta Raymond P. , Bradley Dorothy G. , McDonald Kerry M. , Tökés László
Published: January 2020

dl
-9-Oxo-13 (
cis
and
trans
)-prostenoic acids were converted in high yields into their 18- and 19-hydroxy derivatives by cultures of
Microascus trigonosporus.
The structure elucidation of the microbial products is described.
Virus inactivation by grapes and wines
Authors: Konowalchuk J , Speirs J I
Published: January 2020

Infusions and extracts of different grapes inactivated poliovirus; agents responsible for this property resided in the skin of the grape. Commercial grape juice at both natural and neutral pH inactivate various enteric viruses and herpes simplex virus; a 1,000-fold reduction in poliovirus infectivity occurred after incubation with grape juice, pH 7.0, for 24 h at 4 degrees C. A variety of wines were antiviral but to a lesser extent than grape juice; red wines were more antiviral than white. Antiviral activity was demonstrable in fractions of grape juice varying in molecular weight from less than 1,000 to greater than 30,000 as determined by membrane filtration. Some restoration of poliovirus infectivity from virus-grape juice complexes was achieved with 1% gelatin, 0.1% Tween 80, 0.5% polyvinyl pyrrolidone, and 0.5% polyethylene glycol.
Microbial oxidation and assimilation of propylene
Authors: Cerniglia C E , Blevins W T , Perry J J
Published: January 2020

Hydrocarbon-utilizing microorganisms in our culture collection oxidized propylene but could not utilize it as the sole source of carbon and energy. When propane-grown cells of Mycobacterium convulutum were placed on propylene, acrylate, the terminally oxidized, three-carbon unsaturated acid, accumulated. A mixed culture and an axenic culture (strain PL-1) that utilized propylene as the sole source of carbon and energy were isolated from soil. Respiration rates, enzyme assays, fatty acid profiles, and 14CO2 incorporation experiments suggest that both the mixed culture and strain PL-1 oxidize propylene via attack at the double bond, resulting in a C2+C1 cleavage of the molecule.
Nitrate removal in closed-system aquaculture by columnar denitrification
Authors: Balderston W L , Sieburth J M
Published: January 2020

The columnar denitrification method of nitrate-nitrogen removal from high-density, closed system, salmonid aquaculture was investigated and found to be feasible. However, adequate chemical monitoring was found to be necessary for the optimization and quality control of this method. When methanol-carbon was not balanced with inlet nitrate-nitrogen, the column effluent became unsatisfactory for closed-system fish culture due to the presence of excess amounts of nitrite, ammonia, sulfide, and dissolved organic carbon. Sulfide production was also influenced by column maturity and residence time. Methane-carbon was found to be unsatisfactory as an exogenous carbon source. Endogenous carbon could not support high removal efficiencies. Freshwater columns adpated readily to an artificial seawater with a salinity of 18% without observable inhibition. Scanning electron microscopy revealed that the bacterial flora was mainly rod forms with the Peritricha (protozoa) dominating as the primary consumers. Denitrifying bacteria isolated from freshwater columns were tentatively identified as species of Pseudomonas and Alcaligenes. A pilot plant column was found to behave in a manner similar to the laboratory columns except that nitrite production was never observed.
Enteropathogenicity of Vibrio parahaemolyticus in the ligated rabbit ileum
Authors: Brown D F , Spaulding P L , Twedt R M
Published: January 2020

The enteropathogenicity of Vibrio parahaemolyticus was investigated by contrasting the effects of whole cells, cell fragments, cell-free preparations, and media constituents injected into rabbit ileal loops. Three of 20 cultures utilized were Kanagawa-negative strains from seawater and sea fish. The remaining 17 cultures included both Kanagawa-positive and -negative strains from Japanese victims of gastroenteritis. Broth culture filtrates concentrated 10-fold by dialysis against 30% Carbowax were unreactive, whereas lyophilized filtrates, regardless of Kanagawa type, as well as all sterile broth preparations containing greater than or equal to 5% NaCl gave positive reactions in the rabbit gut. In contrast, crude lysates derived from broth cultures of Kanagawa-positive strains caused loop dilatation; lysate supernatants were unreactive. Lysates of cells washed from brain heart infusion agar were more reactive than lysates from Trypticase soy agar-grown cells. When agar-grown cell lysates prepared by disruption in saline were dialyzed against distilled water, they were devoid of gut reactivity. Reactivity was restored in dialysands resuspended in saline and in dialysates concentrated 10-fold. The agar-grown cell lysates exhibited Kanagawa-type hemolysis. Our data support the conclusion that the rabbit loop reactivity observed with lyophilized, cell-free culture filtrates may result from excessively elevated NaCl concentrations, and that a toxic factor associated with large-cell particles may be dialyzable, depends on saline for expression, and resembles the Kanagawa hemolysin.
Aeromonas
Distribution and Survival in a Thermally Altered Lake
Authors: Fliermans C. B. , Gorden R. W. , Hazen T. C. , Esch G. W.
Published: January 2020

Par Pond is a thermally enriched monomictic southeastern lake which receives heated effluent from a production nuclear reactor. Fish populations in the lake have lesions of epizooty from which
Aeromonas
spp. are readily isolated. Distribution and population densities of
Aeromonas
in the water column were measured along an oxygen and temperature gradient as well as seasonally. Greater population densities of
Aeromonas
occurred below the oxygen chemocline when the lake was stratified. Survival of
Aeromonas hydrophila
under in situ conditions in both epilimnetic and hypolimnetic waters was determined through the use of polycarbonate membrane diffusion chambers during two separate reactor operating conditions. Survival levels of pure cultures of
A. hydrophila
corresponded to the distribution patterns of the naturally occurring
Aeromonas
-like populations. The greater survival of
A. hydrophila
during full reactor operation suggests that the fish populations may be exposed to
Aeromonas
for a longer period of time than when the reactor is not operating.
Receptor Site on Clover and Alfalfa Roots for
Rhizobium
Authors: Dazzo Frank B. , Brill Winston J.
Published: January 2020

Sites on white clover and alfalfa roots that bind
Rhizobium trifolii
and
R. meliloti
capsular polysaccharides, respectively, were examined by fluorescence microscopy. Fluorescein isothiocyanate-labeled capsular material from
R. trifolii
bound specifically to root hairs of clover but not alfalfa. Binding was most intense at the root hair tips. Treatment of clover roots with 2-deoxyglucose (2-dG) prevented binding of
R. trifolii
capsular material to the roots. The sugar 2-dG enhanced the elution of clover root protein, which could bind to and specifically agglutinate
R. trifolii
but not
R. meliloti
or
R. japonicum
. The mild elution procedure left the roots intact. Agglutination of
R. trifolii
and passive hemagglutination of rabbit erythrocytes coated with the capsular material of
R. trifolii
were specifically inhibited by 2-dG. These results suggest that clover roots contain proteins that cross-link complementary polysaccharides on the surface of clover root hairs and infective
R. trifolii
through 2-dG-sensitive binding sites. Alfalfa root hairs were shown to specifically bind to a surface polysaccharide from
R. meliloti.
Aggregation of poliovirus and reovirus by dilution in water
Authors: Floyd R , Sharp D G
Published: January 2020

Poliovirus and reovirus were found to aggregate into clumps of up to several hundred particles when diluted 10-fold into distilled water from a stock preparation of minimal aggregation in 0.05 M phosphate buffer, pH 7.2, plus 22 to 30% sucrose. Reovirus was also found to aggregate when diluted into phosphate-buffered saline. The aggregation was concentration dependent and did not occur when either virus was diluted into water 100-fold or greater. The aggregation of poliovirus was reversible by further addition of saline and produced a dispersed preparation of virus. Reovirus aggregation was not reversible. Both viruses aggregated when diluted into buffers at pH 5 and 3, and poliovirus aggregated at pH 6, and this aggregation of both viruses was reversible when returned to pH 7. Aggregation did not occur at alkaline pH values. Aggregation at low pH could be caused aggregation of either virus at pH 7. Calcium ions, however, were found to aggregate both viruses at a concentration of 0.01 M.
Poliovirus aggregates and their survival in water
Authors: Young D C , Sharp D G
Published: January 2020

Inactivation of aggregated poliovirus by bromine is characterized by a continuously decreasing reaction rate. Poliovirus released from infected cells in these experiments by alternate freezing and thawing in water without electrolytes has always been aggregated. The aggregates persist even on 7,000-fold dilution in ion-free water. Virus similarly released into phosphate-buffered saline solution may be well dispersed, but it aggregates when sedimented into a salt-free sucrose gradient or when it is diluted as little as 10-fold in water. Large one-step dilutions of dispersed virus in water remain dispersed. Aggregated virus was not dispersed by one-step dilution (7,000-fold) in distilled or untreated lake water but was dispersed if phosphate-buffered saline or clarified secondary sewage plant effluent was used as diluent. Dispersed virus aggregates at all dilutions in alum-treated, finished water from the city filter plant. This may be the result of complex formation with insoluble material rather than virion-virion aggregation. A simple procedure is described for rendering a very dilute suspension of mixed virion aggregates into a three-part spectrum of sizes.
Coliforms from hides and meat
Authors: Newton K G , Harrison J C , Smith K M
Published: January 2020

Coliform tests were performed on 85 hide and 75 meat samples. IMViC reactions were determined on isolates from positive confirmed and fecal tests, and strains other than Escherichia coli were identified. Strains typed as Aerobacter aerogenes types I and II were identified as Enterobacter cloacae (51.4%), Klebsiella pneumoniae (21.5%), Enterobacter aerogenes (15%), and Enterobacter liquefaciens, Serratia, and unidentified coliforms (12.1%). K. pneumoniae appeared to be responsible for less than 1% positive fecal tests.
Slide staining device for use during space flight
Authors: Brockett R M , Brady J , Day J L , Ferguson J K , Walsh J M
Published: January 2020

A slide staining device is described that performs Gram and Wright stains during space flight. Reagents and liquid wastes are contained within a closed system.
Mutant of Aspergillus flavus producing more aflatoxin B2 than B1
Authors: Papa K E
Published: January 2020

A mutant of Aspergillus flavus having a high and relatively stable aflatoxin B2/B1 ratio was recovered after treatment with nitrosoguanidine.
Chemically defined medium for Rothia dentocariosa
Authors: Lesher R J , Deal S J
Published: January 2020

A chemically defined medium for Rothia dentocariosa has been established. All strains tested, including the type strain, grew well in this medium. Rothia strains had a generation time of 3.5 to 4.0 h, with a yield of 62 mg (dry weight) per 100 ml after 72 h of growth in the chemically defined medium.
Comparison of microbial counts on beef carcasses by using the moist-swab contact method and secondary tissue removal technique
Authors: Lazarus C R , Abu-Bakar A , West R L , Oblinger J L
Published: January 2020

When a tissue removal rinse technique was compared to the moist-swab contact method, significantly greater numbers of bacteria were recovered from beef carcasses, especially when the flora exceeded log10 4.5/6.45 cm2. Secondary treatment of the removed surface tissue by blending resulted in a significantly greater number of bacteria being recovered than when the same sample was swabbed and/or rinsed. Data indicate that blending of the carcass surface tissue provides a more representative value of the true microbial flora.
Fermentation of L-aspartate by a saccharolytic strain of Bacteroides melaninogenicus
Authors: Wong J C , Dyer J K , Tribble J L
Published: January 2020

Resting cells of Bacteroides melaninogenicus fermented L-[14C]aspartate as a single substrate. The 14C-labeled products included succinate, acetate, CO2, oxaloacetate, formate, malate, glycine, alanine, and fumarate in the relative percentages 68, 15, 9.9, 2.7, 1.8, 1.0, 0.7, 0.5, and 0.06, respectively, based on the total counts per minute of the L-[14C]aspartate fermented. Ammonia was produced in high amounts, indicating that 96% of the L-aspartate fermented was deaminated. These data suggest that L-aspartate is mainly being reduced through a number of intermediate reactions involving enzymes of the tricarboxylic acid cycle to succinate. L-[14C]asparagine was also fermented by resting cells of B. melaninogenicus to form L-aspartate, which was subsequently, but less actively, fermented.
Isolation and identification of fecal bacteria from adult swine
Authors: Salanitro J P , Blake I G , Muirhead P A
Published: January 2020

An examination of the fecal microflora of adult swine was made with regard to the efficiency of several roll tube media in enumeration and recovery of anaerobes, the effects of medium constituents on recovery, and the isolation and identification of the predominant kinds of bacteria. Total number of organisms by microscopic bacterial counts varied among fecal samples from 4.48 X 10(10) to 7.40 X 10(10) bacteria/g (wet weight). Comparison of different nonselective roll tube media indicated that about 30% of the fecal bacteria could be recovered with a rumen fluid (40%, vol/vol) medium (M98-5). Recoveries of 21 and 15%, respectively, were obtained with M10 and rumen fluid-glucose-cellobiose agar (RGCA) media. Rumen fluid, Trypticase, sugars, and CO2 gas phase were important components required for maximum recovery with this medium. Similar high recoveries of anaerobes were also obtained with M98-5 containing swine cecal extract of place in rumen fluid or M10 plus swine cecal extract. Significantly lower recoveries were observed with RCGA, media supplemented with swine fecal extracts, reinforced clostridial medium, brain heart infusion agar, and prereduced blood agar. Ninety percent of the bacteria isolated from roll tube media were gram positive and consisted of facultatively anaerobic streptococci, Eubacterium sp., Clostridium sp., and Propionibacterium acnes. The remainder of the flora (8%) included several other species of anaerobes and Escherichia coli. Rumen fluid (or volatile fatty acids), Trypticase, and yeast extract additions to basal media stimulated the growth of anaerobic strains. Variation in the relative proportions of the predominant fecal microflora was observed. This work indicates that satisfactory enumeration, isolation and cultivation of the predominant microflora in swine feces can be obtained when strict anaerobic culture methods and a rumen fluid medium are used.