bluuebunny/crossref_metadata_2025 huggingface dataset:
Lethal cold stress of Vibrio vulnificus in oysters
Authors: Oliver J D
Published: January 2020

Studies were conducted on the survival of Vibrio vulnificus, an estuarine human pathogen, in oyster homogenates held at 4 degrees C. Results indicated a rapid and dramatic decrease in viability not attributable to either cold shock or the oyster homogenate alone but to a combination of the two. Such a decline was not observed with Vibrio parahaemolyticus. Chilled V. vulnificus cells were unable to repair themselves in brain heart infusion broth at 37 degrees C. V. vulnificus cells incubated on whole raw oysters at 0.5 degrees C also exhibited a decline in viability, but of a lesser degree. The effects of various plating media were also investigated. The data reported here suggest that oysters kept on ice are not likely to be a major factor in the epidemiology of V. vulnificus infection. It is further suggested that the standard method of homogenizing oysters for examining bacteriological quality should not be followed because toxic compounds are released from the oysters during this process.
Syntrophic association of a butyrate-degrading bacterium and methanosarcina enriched from bovine rumen fluid
Authors: McInerney M J , Mackie R I , Bryant M P
Published: January 2020

An anaerobic butyrate-degrading bacterium, morphologically similar to Syntrophomonas wolfei, was isolated in coculture with Desulfovibrio strain G11 from an enrichment of bovine rumen fluid. A Methanosarcina species was the major H2-using organism in the enrichment. The results are discussed in relationship to the absence of Methanospirillum hungatei, the H2-using methanogen usually found in association with S. wolfei, and the finding of Methanosarcina rather than Methanobrevibacter ruminantium as the major H2-using bacterium in the enrichments. The finding of butyrate degraders in the rumen suggests that, if the retention time of the rumen contents becomes more prolonged, butyrate and longer-chained fatty acids might be significantly degraded.
Microbial transformations of 7,2-dimethylbenz[a]anthracene
Authors: Wu J , Wong L K
Published: January 2020

Microbial transformations of 7,12-dimethylbenz[a]anthracene, a carcinogenic polycyclic aromatic hydrocarbon, in cultures of Pseudomonas aeruginosa and Penicillium notatum were studied by high performance liquid chromatographic separation of metabolic fractions followed by gas chromatographic-mass spectrometric analysis of the metabolites. Two methyl-hydroxylated metabolites were identified in each of the incubations. The metabolic activation of the polycyclic aromatic hydrocarbon suggests a possible involvement of microorganisms in environmental carcinogenesis.
Enumeration of mycoplasmas after acridine orange staining
Authors: Rosendal S , Valdivieso-Garcia A
Published: January 2020

Samples of liquid mycoplasma cultures were mixed with equal part of a 0.01% solution of acridine orange and placed on agar plates. The number of fluorescing organisms per field was counted in an epifluorescence microscope at an X 1,000 magnification. When the number of fluorescing organisms per field was related to the number of colony-forming units per milliliter during the growth cycle, highly significant correlation was found in cultures with greater than or equal to 10(6) colony-forming units per ml during the exponential growth phase. The counts were weakly correlated during the stationary phase and not correlated during the death phase. This technique provides a mean to enumerate mycoplasmas in liquid cultures.
Isolation and Presumptive Identification of Adherent Epithelial Bacteria (“Epimural” Bacteria) from the Ovine Rumen Wall
Authors: Mead Lorna J. , Jones G. A.
Published: January 2020

One hundred sixty-one strains of adherent bacteria were isolated under anaerobic conditions from four sites on the rumen epithelial surface of sheep fed hay or a hay-grain ration. Before isolation of bacteria, rumen tissue was washed six times in an anaerobic dilution solution, and viable bacteria suspended in the washings were counted. Calculation indicated that unattached bacteria would have been removed from the tissue by this procedure, but a slow and progressive release of attached bacteria also occurred. Nevertheless, a wide range of characteristic morphological types remained associated with the epithelium as demonstrated by scanning electron microscopy. Most of these types were represented among the isolates. Characterization and presumptive identification of the isolates showed that 95.0% belonged to previously described genera of functionally significant rumen bacteria, including
Butyrivibrio
sp. (31.1%),
Bacteroides
sp. (22.4%),
Selenomonas ruminantium
(9.9%),
Succinivibrio dextrinosolvens
(8.7%),
Streptococcus bovis
(8.1%),
Propionibacterium
sp. (4.3%),
Treponema
sp. (3.1%), and
Eubacterium
sp.,
Lachnospira multiparus
, and
Ruminococcus flavefaciens
(2.5% each). Eight isolates (5.0%) were not identified.
L. multiparus
was recovered only from hay-fed animals; all other genera were obtained from animals fed either ration. All
S. bovis
strains and two strains each of
Bacteroides
sp. and
Butyrivibrio
sp. were aerotolerant; all other strains were strictly anaerobic. Bacteria representing the gram-positive, facultatively anaerobic flora associated with rumen wall tissue (R. J. Wallace, K.-J. Cheng, D. Dinsdale, and E. R. Ørskov, Nature (London)
279
:424-426, 1979) were therefore not recovered by the techniques used; instead a different fraction of the adherent population was isolated. The term “epimural” is proposed to describe the flora associated with the rumen epithelium.
Thermal inactivation of infectious hematopoietic necrosis and infectious pancreatic necrosis viruses
Authors: Gosting L H , Gould R W
Published: January 2020

A plaque assay was used to follow the inactivation kinetics of infectious hematopoietic necrosis virus and infectious pancreatic necrosis virus in cell culture media at various temperatures. Inactivation of infectious hematopoietic necrosis virus in a visceral organ slurry was compared with that in culture media.
Enterobacteriaceae associated with meats and meat handling
Authors: Stiles M E , Ng L K
Published: January 2020

The source of Enterobacteriaceae on meats was shown to be associated with the meat-handling work surfaces in two packing plants studies. A total of 2,343 Enterobacteriaceae were isolated and identified from meat samples and work surfaces at the packing plants and at the retail facilities. Escherichia coli biotype I and Serratia liquefaciens were detected at all stages of meat handling, indicating that they may be present in meats throughout the meat-handling system. Enterobacter agglomerans and S. liquefaciens were the predominant Enterobacteriaceae at the retail level, but they had limited indicator potential for sanitation and hygiene, Klebsiella pneumoniae was a frequent isolate among Enterobacteriaceae from meats and meat-handling surfaces in the packing plants but not at the retail level, indicating that this organism might signal unhygienic handling of meats at the retail level.
Effects of Pharmaceutical Wastes on Microbial Populations in Surface Waters at the Puerto Rico Dump Site in the Atlantic Ocean
Authors: Peele E. R. , Singleton F. L. , Deming J. W. , Cavari B. , Colwell R. R.
Published: January 2020

A series of cruises during 1979 and 1980 to the pharmaceutical dump site located 64 km north of Arecibo, Puerto Rico, in the Atlantic Ocean, was carried out to evaluate effects of wastes on the ecology of the microflora of surface waters of the dump site. In addition to bacteriological monitoring of the waste plume created by the release of wastes from the disposal barge, stations along a series of transects, extending north from coastal waters through and beyond the dump site, were sampled. Largest numbers of culturable bacteria on marine agar were found at stations closest to shore and in the vicinity of the dump site. Bacteria recovered on marine agar were predominantly
Vibrio
and
Aeromonas
spp., with the relative abundance of these organisms decreasing as gram-positive organisms (staphylococci, micrococci, and bacilli) became dominant in areas immediately affected by waste dumping. Total numbers of bacteria (determined by acridine orange direct counts [AODC]), which were relatively stable throughout the region, and a direct estimate of viable cells (DVC), i.e., those cells responsive to additions of yeast extract and nalidixic acid, were determined by acridine orange staining and epifluorescence microscopy. Heterotrophic bacterial activity, measured by the uptake (
V
max
) of
14
C-labeled amino acids, declined relative to distance from land. Increases in specific activity indices (DVC/AODC and
V
max
/AODC) were observed near the dump site. The composite results of this study, i.e., increased specific activities (determined by two methods), increased numbers of culturable marine bacteria, and marked alteration of the taxonomic composition of the culturable bacterial community in waters within and surrounding the Puerto Rico dump site, indicate demonstrable changes in the marine microbial community in the region used for waste disposal.
Lectin-Like Constituents of Foods Which React with Components of Serum, Saliva, and
Streptococcus mutans
Authors: Gibbons R. J. , Dankers I.
Published: January 2020

Hot and cold aqueous extracts were prepared from 22 commonly ingested fruits, vegetables, and seeds. When tested by agar diffusion, extracts from 13 and 10 of the foods formed precipitin bands with samples of normal rabbit serum and human saliva, respectively; extracts from four of the foods also reacted with antigen extracts of strains of
Streptococcus mutans
. When added to rabbit antiserum, extracts from 18 of 21 foods tested inhibited reactivity with antigen extracts derived from
S. mutans
MT3. Extracts from 16 foods agglutinated whole
S. mutans
cells, whereas those from 10 foods agglutinated human erythrocytes of blood types A and B. The lectin-like activities of extracts which reacted with human saliva were studied further. Pretreatment of saliva-coated hydroxyapatite (S-HA) beads with extracts of bananas, coconuts, carrots, alfalfa, and sunflower seeds markedly reduced the subsequent adsorption of
S. mutans
MT3. Pretreatment of S-HA with banana extract also strongly inhibited adsorption of
S. mutans
H12 and
S. sanguis
C1, but it had little effect on attachment of
Actinomyces naeslundii
L13 or
A. viscosus
LY7. Absorption experiments indicated that the component(s) in banana extract responsible for inhibiting streptococcal adsorption to S-HA was identical to that which bound to human erythrocytes. The banana hemagglutinin exhibited highest activity between pH 7 and 8, and it was inhibited by high concentrations of glucosamine, galactosamine, and, to a lesser extent, mannosamine. Other sugars tested had no effect. The selective bacterial adsorption-inhibiting effect noted for banana extract was also observed in studies with purified lectins. Thus, pretreating S-HA with wheat germ agglutinin and concanavalin A inhibited adsorption of
S. mutans
MT3 cells, whereas peanut agglutinin,
Ulex
agglutinin,
Dolichos
agglutinin, and soybean agglutinin had little effect; none of these lectins affected attachment of
A. viscosus
LY7. Collectively, the observations suggest that many foods contain lectins which can interact with components of human saliva and
S. mutans
cells. Because of their potential to influence host-parasite interactions in the mouth and elsewhere in the gastrointestinal canal, these reactions warrant further study.
Incidence of lysogeny, colicinogeny, and drug resistance in enterobacteria isolated from sewage and from rectum of humans and some domesticated species
Authors: Dhillon T S , Dhillon E K
Published: January 2020

Enterobacteria were isolated by streaking swabs of sewage and rectal swabs from human volunteers from domesticated animals. Thirty strains of human origin were identified as Escherichia coli. Out of 1,367 rectal isolates of animal origin, 21% were lysogenic (phi+), 29% were colicinogenic (col+), and 7% were col+ phi+. Out of 85 rectal samples more than 60% harbored variable numbers of col+ or phi+ bacteria. Lysogens harboring homoimmune prophages were detectable in six out of eight human subjects in sequential samples taken at weekly intervals. Chickens in Hong Kong are fed on antibiotic-containing feeds; the avian isolates contained the highest frequency (98%) of drug-resistant bacteria, whereas only 39% of the bovine and 61% of the human isolates were drug resistant. Transmissible drug resistance was demonstrable in sewage isolates and those from animal sources; the highest frequency (58%) of resistance donors was shown by the avian isolates, and the lowest (9%) was shown by the bovine isolates. Unselected marker analysis has shown that a vast majority of multiply resistant donors of diverse origins are able to transmit multiple resistance.
Sensitive and Accurate Methodology for Measuring the Kinetics of Concentration-Dependent Hydrocarbon Metabolism Rates in Seawater by Microbial Communities
Authors: Button D. K. , Schell D. M. , Robertson B. R.
Published: January 2020

A method having sufficient sensitivity to resolve the kinetic constants for dissolved nonpolar substrate metabolism, together with the related rate constants in natural waters, is presented. The method is based on the rate of
14
CO
2
recovery from radioactive dissolved substrate. Sensitivity is enhanced by using large seawater volumes, high-specific-activity isotopes, and by reducing background radioactivity. Before use, commercial isotopes are purified by mild alkaline hydrolysis followed by sublimation from base to remove
14
CO
2
as well as interfering polar
14
C-substrates. During sample analysis, chilled Tenax resin is used to remove volatile
14
C-substrate from the nitrogen stream containing
14
CO
2
recovered from substrate oxidation. Chromatographic evidence of purity, shown to be insufficient, is augmented by kinetic data from toluene utilization by mixed cultures and by rates in induced versus noninduced pure cultures. Accuracy is enhanced by using short (<10 h) incubation times and small hydrocarbon concentrations so that the metabolism rates in unamended natural water systems can be evaluated. Toluene metabolism rates in seawater as low as 1 pg/liter per h and at concentrations as low as 20 ng/liter have been determined.
New selective agent for isolation of Pseudomonas aeruginosa
Authors: Marold L M , Freedman R , Chamberlain R E , Miyashiro J J
Published: January 2020

Results of minimal inhibitory concentration tests with a diversity of bacterial strains showed that 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan (C-390) inhibited the growth of all microorganisms tested (other than Pseudomonas aeruginosa) at 25 microgram/ml or less, whereas MICs obtained for P. aeruginosa ranged from to to greater than 100 microgram/ml. Therefore, C-390 was evaluated as a potential selective agent for P. aeruginosa in pseudomonas agar F. Recovery tests were conducted on this medium with 53 strains o P. aeruginosa, and the results were compared to those obtained in similar tests on commercially available selective media, i.e., pseudomonas isolation agar and Pseudosel agar. The results of these comparisons indicated that pseudomonas agar F with C-390 was significantly less inhibitory than Pseudosel agar and pseudomonas isolation agar and more selective than pseudomonas isolation agar. The incorporation of C-390 in pseudomonas agar F also provided a medium that was both selective and differential. Preliminary evidence also suggested that C-390 may be added to other basal media with comparable results.
Heat inactivation of staphylococcus epidermidis at various water activities
Authors: Verrips C T , Van Rhee R
Published: January 2020

Members of the family Micrococcaceae play an important role in food spoilage and even in food poisoning. In contrast to members of the family Enterobacteriaceae, these bacteria can grow in media with low water activities. Therefore, the heat resistance of Staphylococcus epidermidis, a rather resistant member of the family Micrococcaceae, was studied at water activities between 0.87 and 1.00. The heat inactivation curves were clearly biphasic at all temperatures and water activities tested. Especially at low water activities, the D-values of the tail phase were extremely high (at 0.87 water activity, a D-value at 70 degrees C of 500 s was recorded).
Impact of verification media and resuscitation on accuracy of the membrane filter total coliform enumeration technique
Authors: Evans T M , Seidler R J , LeChevallier M W
Published: January 2020

Verification of membrane filter total coliform colonies was compared in lauryl tryptose broth, and m-LAC broth primary media and brilliant green-lactose-bile broth and EC broth secondary media. Verification in m-LAC broth yielded the greatest number of aerogenic isolates for both untreated surface water and drinking water samples. Verification in brilliant green-lactose-bile broth increased the number of false-negative reactions. At least 90% of the isolates aerogenic in primary verification media and anaerogenic in brilliant green-lactose-bile broth were representative of the coliform genera. The addition of a resuscitation step in the membrane filter technique did not yield greater numbers of verified coliforms per sample. Verification of both typical and atypical colonies in m-LAC broth resulted in a 10-fold increase in coliform numbers from untreated surface water. With drinking water, verification of both colony types resulted in an increase from less than 1 coliform per 100 ml to greater than 1/100 ml. A single-step verification in m-LAC broth is proposed as a more rapid and sensitive coliform verification procedure than the standard technique.
Inhibition of catechol 2,3-dioxygenase from Pseudomonas putida by 3-chlorocatechol
Authors: Klecka G M , Gibson D T
Published: January 2020

Partially purified preparations of catechol 2,3-dioxygenase from toluene-grown cells of Pseudomonas putida catalyzed the stoichiometric oxidation of 3-methylcatechol to 2-hydroxy-6-oxohepta-2,4-dienoate. Other substrates oxidized by the enzyme preparation were catechol, 4-methylcatechol, and 4-fluorocatechol. The apparent Michaelis constants for 3-methylcatechol and catechol were 10.6 and 22.0 muM, respectively. Substitution at the 4-position decreases the affinity and activity of the enzyme for the substrate. Catechol 2,3-dioxygenase preparations did not oxidize 3-chlorocatechol. In addition, incubation of the enzyme with 3-chlorocatechol led to inactivation of the enzyme. Kinetic analyses revealed that both 3-chlorocatechol and 4-chlorocatechol were noncompetitive or mixed-type inhibitors of the enzyme. 3-Chlorocatechol (Ki = 0.14 muM) was a more potent inhibitor than 4-chlorocatechol (Ki = 50 muM). The effect of the ion-chelating agents Tiron and o-phenanthrolene were compared with that of 3-chlorocatechol on the inactivation of the enzyme. Each inhibitor appeared to remove iron from the enzyme, since inactive enzyme preparations could be fully reactivated by treatment with ferrous iron and a reducing agent.
Carbon Monoxide Metabolism in Roadside Soils
Authors: Spratt Henry G. , Hubbard Jerry S.
Published: January 2020

Air-dried soils which were equilibrated under relative humidities greater than 93% or moistened with liquid water showed marked increases in their capacities to oxidize CO to CO
2
. Liquid water addition in excess of saturation resulted in lower CO oxidation rates, reflecting the limited diffusion of CO through the aqueous phase. After 35 days' storage under 100% relative humidity, the capacity for CO oxidation decreased to 21% of the value observed with a freshly collected sample. Incubation of this stored soil under an atmosphere containing 200 ppm of CO (250 mg/m
3
) for 21 days resulted in a sevenfold increase in CO oxidation. A correlation was noted between the CO oxidative activity and the history of previous exposure of soils to high ambient levels of CO. The organisms responsible for CO oxidation apparently comprise a small fraction of the microbial population in the soils. With a roadside soil the oxidation of CO provided the driving force for the assimilation of CO
2
. The stoichiometry of the oxidative and assimilatory reactions in soil was in the range of values reported from laboratory studies with CO chemoautotrophs (carboxydobacteria). It is proposed that the population and activity of CO-oxidizing microorganisms increase in response to increasing levels of CO in the environment.
Isolation of
Legionella pneumophila
from Cooling Tower Water by Filtration
Authors: Orrison Leta H. , Cherry William B. , Milan David
Published: January 2020

Methods are described for detection of
Legionella pneumophila
in cooling tower water or other water sources by direct fluorescent-antibody staining. A procedure for isolation of
Legionella
bacteria from water samples by guinea pig inoculation is described. Two different serogroups of
L. pneumophila
were isolated repeatedly from one of the cooling towers.
Particle size of airborn Cryptococcus neoformans in a tower
Authors: Ruiz A , Bulmer G S
Published: January 2020

Nearly 10(6) cells of Cryptococcus neoformans were cultured per g of pigeon droppings in a vacant tower. The air in the tower contained an average of 45 viable cells of C. neoformans per 100 liters: 60% of the cells were less than 4.7 micron in diameter. It is estimated that a human exposed to this atmosphere for 1 h would have 41 cells of c. neoformans deposited in the lungs. Sweeping resulted in the aerosolization of large numbers of cells of C. neoformans from 4.7 to 11 micron in diameter, the number of cells less than 4.7 micron remained relatively constant. One minute after sweeping, 4.4% of viable airborne cells of C. neoformans were less than 1.1 micron in diameter. We believe that this is the first report of isolating such small cells of C. neoformans from a natural site.
Activation of Polynuclear Aromatic Hydrocarbons to Mutagens by the Marine Ciliate
Parauronema acutum
Authors: Lindmark Donald G.
Published: January 2020

The marine ciliate
Parauronema acutum
converted 2-aminofluorene and 2-acetylaminofluorene to compounds with mutagenic activity in the Ames
Salmonella
test. The ciliate, however, did not activate benzo (α)pyrene or benzanthracene or destroy the mutagenic properties of nitrosoguanidine. Homogenates, when substituted for the liver S-9 fraction in the
Salmonella
/microsome test, activated 2-aminofluorene and 2-acetylaminofluorene to mutagens. Benzo(α)pyrene and benzanthracene were not activated, nor was nitrosoguanidine inactivated. Phenobarbitol did not induce or increase the amount of activating activity. The activation showed no requirement for the reduced nicotinamide adenine dinucleotide phosphate-regenerating system required by liver P-450 cytochromes. Upon differential sedimentation of a cell homogenate, the majority of the activity sedimented with a small-particle fraction with sedimentation properties like those of microsomes from higher eucaryotes. Benzo(α)pyrene, although not metabolized, was accumulated by cultures of
P. acutum
at a linear rate and was not appreciably released (10%) after removal of benzo(α)pyrene from the incubation medium. Hence, this ciliate could convert certain polynuclear aromatic hydrocarbons to mutagens and accumulate others.
Fungal Metabolite from Members of the Genus
Rhizopus
Authors: Thompson D. P. , Smalley A. W. , Eribo Broderick E.
Published: January 2020

A fluorescent metabolite present in seven members of the genus
Rhizopus
was isolated. This compound appeared green before spray treatment and purple after spray treatment with
p
-anisaldehyde in visible light. Subsequent purification and structural elucidation of the isolated compound yielded 1-[2,6,10,14-tetramethyl-17-carbomethyl heptadecyl]-1-[2,6,10,14-tetramethyl-17-methanoyl heptadecyl]-benzene.
Reduction of Polymeric Azo and Nitro Dyes by Intestinal Bacteria
Authors: Brown Joseph P.
Published: January 2020

The O
2
-sensitive reduction of high-molecular-weight aromatic azo and nitro dyes by intestinal bacteria appears to be mediated by low-molecular-weight electron carriers with
E
o
′ = −200 to −350 mV. This process may allow the design of polymeric azo prodrugs for specific release of certain aromatic amines in the colon.
Detection of
Escherichia coli
in Foods: Indole Staining Methods for Cellulosic and Polysulfone Membrane Filters
Authors: Sharpe Anthony N. , Peterkin Pearl I. , Rayman M. Khalil
Published: January 2020

Optimized procedures for staining
Escherichia coli
colonies on cellulosic and polysulfone membrane filters are described. An explanation for the behavior of the Ehrlich reaction on membrane filters is suggested.
Resistance of selected strains of Pseudomonas aeruginosa to low-intensity ultraviolet radiation
Authors: Abshire R L , Dunton H
Published: January 2020

The resistances of 10 strains of Pseudomonas aeruginosa and other microorganisms to an ultraviolet (UV) intensity of 100 muW/cm2 were determined. Organisms were exposed in 2- or 15-ml saline suspensions contained in uncapped polyethylene bottles for increasing periods of time, and the surviving fractions were enumerated. Decimal reduction times were calculated by regression analysis, using the least-squares method. The 10 strains of P. aeruginosa, compared with Micrococcus radiodurans and Candida albicans, were very susceptible to low-intensity UV radiation. Results from this study showed that a UV intensity of 100 muW/cm2 penetrated saline suspensions up to 40 mm deep sufficiently to kill high levels of microbial cells, especially P. aeruginosa cells. These results allowed us to design a system for determining and monitoring the sterilization capability of low-intensity UV radiation. In our particular case, UV proved to be an efficient mode for sterilizing saline suspensions of P. aeruginosa in polyethylene bottles. The significance and application of these findings with regard to supporting UV as a sterilant are discussed.
Attachment of
Streptococcus faecium
to the Duodenal Epithelium of the Chicken and Its Importance in Colonization of the Small Intestine
Authors: Fuller R. , Houghton S. B. , Brooker B. E.
Published: January 2020

The counts of
Streptococcus faecium
SY1 in the duodenums of gnotobiotic chicks exceeded the counts in their crops, indicating that multiplication was occurring in the anterior small intestine. This growth was related to adhesion to the gut wall which could be demonstrated by viable counts of macerated washed duodenal tissue. Scanning electron microscopy demonstrated that adhesion occurred in restricted areas on the surface of the villus, and transmission studies showed the presence of a thick extracellular layer on the bacterium. Attachment of
S. faecium
SY1 was confirmed in vitro by using chicken duodenal brush borders. The washings, produced during the preparation of the brush borders, increased the number of
S. faecium
adhering to the brush borders. This enhancing effect was due to the presence of trypsin in the duodenal washings. However, the effect was not dependent on the enzymatic activity of the trypsin molecule. The initial adhesion was not prevented by pretreatment of the brush borders with soy bean trypsin inhibitor. There were, therefore, two adhesion systems operating, only one of which was dependent on trypsin. Pretreatment of brush borders with trypsin digested them, but they remained intact in the presence of
S. faecium
SY1, indicating that the enzymatic activity was being inhibited. This effect was specific for the adhering strain of
S. faecium
SY1; the nonadhering
S. faecium
strain CRS23 and an adhering strain of
Lactobacillus
sp. were inactive, as was strain SY1 when adhesion was prevented by including sodium periodate in the test system. The colonizations of the gut by strains of
S. faecium
of differing adhesive abilities were compared. The nonadhering strain CRS23 showed reduced ability to colonize the duodenum, but the penicillin-resistant mutant of
S. faecium
SY1, which had reduced adhesive ability but could still attach to a lesser degree, was able to colonize the duodenum as efficiently as the parent strain.
Influence of Temperature, Oxygen, and pH on a Metalimnetic Population of
Oscillatoria rubescens
Authors: Konopka Allan
Published: January 2020

Planktonic
Oscillatoria
spp. often inhabit depths of thermally stratified lakes in which gradients of physical and chemical factors occur. Measurements of photosynthetic rate or photosynthetic carbon metabolism were used to evaluate the importance of vertical gradients of temperature, oxygen, and pH upon
Oscillatoria rubescens
in Crooked Lake, Ind. At the low light intensities experienced in situ, neither photosynthetic rate nor relative incorporation of carbon dioxide into low-molecular-weight compounds, polysaccharide, or protein was affected by temperature. At a 10-fold-higher light intensity, the photosynthetic rate increased as temperature increased; most of the additional carbon accumulated as polysaccharide. Polysaccharide which was synthesized at high light intensity and temperature was respired when the organisms were placed in the dark, but was not used for protein biosynthesis. When
O. rubescens
was shifted from high light to low light, a fraction of the polysaccharide was metabolized into protein. Adaptation to growth at lower temperatures by
O. rubescens
cultures resulted in a decrease in the maximum photosynthetic rate. Oxygen inhibited photosynthesis by only 10 to 15% at concentrations typically found in the lake. The photosynthetic rates at pH values which occurred in Crooked Lake were all near the maximum. Thus, gradients of temperature, oxygen, or pH are not likely to significantly affect the distribution of
O. rubescens
in Crooked Lake, given the low light intensity at which
O. rubescens
grows and the range of values for those factors in the lake.
Specific Lysogenicity in
Streptomyces azureus
Authors: Ogata Seiya , Yoshino Sadazo , Suenaga Hikaru , Aoyama Koji , Kitajima Nakao , Hayashida Shinsaku
Published: January 2020

Slope (or plate) cultures of thiostrepton-producing
Streptomyces azureus
(ATCC 14921) often showed spontaneously developing plaques. Plaques increased in number during serial subcultures. The production of aerial mycelia and sporulating aerial hyphae was interrupted by the overlapping plaques, whereas the growth of substrate mycelia continued in the plaques. These abnormal (eroded) cultures were easily restored to their normal conditions once they were passed through liquid cultures under shaking conditions. A few phage particles were found in the plaques, together with some headless tails and numerous tail tips which formed a hexagonal crystal or a large crystal mass when viewed in an electron microscope. No lytic phenomenon and no phage production were found in the liquid cultures, although all mycelia and spores harbored phage-producing abilities. It was also found that the propagation of phages was successful in solid culture, but not in liquid culture. The whole phage was named SAt2, which belongs to group B of Bradley's morphological classification. From these results, it is considered that
S. azureus
is lysogenic with temperate phage SAt2, of which virulent mutants are able to infect the aerial mycelia and sporulating hyphae of their lysogenic host.
Precursor recognition by kinetic pulse-labeling in a toxigenic aflatoxin B1-producing strain of Aspergillus
Authors: Zamir L O , Hufford K D
Published: January 2020

Kinetic pulse-labeling of aflatoxin pathway compounds was carried out in Aspergillus parasiticus, beginning with radioactive acetate. Norsolorinic acid, averufin, versicolorin A, and sterigmatocystin (all known as compounds which can be incorporated into the aflatoxin molecule) were radiotraced to follow their order of appearance. Aflatoxin species B1, B2, G1, and G2 were included. Norsolorinic acid and averufin appeared as early transient intermediates followed in order by versicolorin A, aflatoxins, and sterigmatocystin. To date, a mutually confirming array of results has been obtained with established precursors in wild-type strains of A. parasiticus and A. versicolor (as well as with an aflatoxin pathway mutant of A. parasiticus), which together establish a practical methodology for recognition of new pathway intermediates. The kinetic of pulse-labeling for sterigmatocystin in relation to aflatoxins suggests that duel branchlets may exist to flatoxins; i.e., sterigmatocystin may not be an obligatory aflatoxin precursor.
Enteropathogenic bacteria in frozen chicken
Authors: Norberg P
Published: January 2020

Eighty-two samples of frozen chicken from retail stores were examined for the presence of Campylobacter, Yersinia enterocolitica, and salmonellae. Aerobic plate counts and numbers of coliform bacteria at 37 degrees C were determined. Campylobacter fetus subsp. jejuni was found in 22% of the samples, Y. enterocolitica was found in 24.5% and Salmonella typhimurium was found in one sample (1.2%). The isolated strains of Y. enterocolitica belonged to serotypes 4, 5b, 6, and 8. Aerobic plate counts and numbers of coliform bacteria at 37 degrees C were not found to be noticeably higher in samples containing pathogens than in pathogen-free samples. This investigation showed that chicken does contain other pathogenic bacteria than salmonellae. Campylobacter and Y. enterocolitica were isolated in much higher frequencies than Salmonella.
Filtering out food debris before microbiological analysis
Authors: Peterkin P I , Sharpe A N
Published: January 2020

Sterile disposable pipette "filter tips" capped with polyethylene mesh (111-microgram pore size) removed bothersome debris from food suspensions before microbiological analysis. A study comprising 576 analyses of Escherichia coli and Staphylococcus aureus in lean and regular ground beef, chicken, chedder and mozzarella cheeses, green and lima beans, rhubarb, and beef and turkey pot pies, showed that these filter tips did not reduce bacterial recovery.
Serum substitute in epithelial cell culture media: nonfat dry milk filtrate
Authors: Fassolitis A C , Novelli R M , Larkin E P
Published: January 2020

A number of milk types and milk fractions were investigated as possible substitutes for serum in cell culture media. A filtrate of reconstituted nonfat dry milk showed promise. Culture fluids containing 5% of the nonfat dry milk filtrate were used to propagate primary and continuous cell cultures, and the cell growth from these cultures was compared with that of cells grown in a serum-containing medium. The nonfat dry milk filtrate-supplemented medium supported the growth of all epithelial cells tested, but two fibroblast-type cultures failed to replicate. Cells grown in the medium containing the milk filtrate grew slowly for 2 to 3 days and then propagated to confluency in 6 to 8 days. Viable cell counts of 9 days were comparable to those of serum-grown cells that had been propagated for 7 days. Cells grown in the milk filtrate could be split 1 to 4 when subcultures were prepared. Cell growth could be stimulated by refeeding on days 2 to 3 or by the addition of 30 microM 2-mercaptoethanol to the growth medium. Virus susceptibility and titer comparisons with poliovirus 1, coxsackievirus B2, echovirus 7, and herpes simplex virus indicated that approximately the same data were obtained when either the nonfat dry milk filtrate-treated or the serum-treated cells were studied. The nonfat dry milk filtrate is inexpensive, is easily prepared, and is a substitute for serum in epithelial cell culture media.
Release of
Rhizobium
spp. from Tropical Soils and Recovery for Immunofluorescence Enumeration
Authors: Kingsley Mark T. , Bohlool B. Ben
Published: January 2020

Limitations associated with immunofluorescence enumeration of bacteria in soil derive largely from the efficiency with which cells can be separated from soil particles and collected on membrane filters for staining. Many tropical soils fix added bacteria tightly, resulting in low recoveries. Eight soils, representative of three of the major soil orders found in the tropics (oxisols, vertisols, and inceptisols), were tested for recovery of added
Rhizobium
strains. All except one Hawaiian andept (Typic Eutrandept) yielded recoveries ranging from <1 to 13%. Recovery from the andept was 100%. In soil-sand mixtures, addition of only a small amount of soil caused a dramatic decrease in recovery of added rhizobia. Increasing the soil content of the mixture from 0% (10 g of sand) to 50% (5 g of soil-5 g of sand) reduced recoveries from >90 to <1%. Varying the ionic strength and pH of the extracting solution did not cause marked increases in recovery. Protein solutions, ethylenediaminetetraacetate, and NaHCO
3
, on the other hand, improved release of bacteria. We report a modification to the usual membrane filter immunofluorescence procedure which yielded consistently high and reproducible recovery (coefficient of variation, 30%) of rhizobia from several tropical soils. In the modified procedure, partially hydrolyzed gelatin, diluted in ammonium phosphate, was used to suspend the soil. This caused dispersion of the soil and release of the bacteria from soil flocs. The efficiency of recovery of
Rhizobium
spp. from several tropical and two temperate soils remained high as the content of these soils in soil-sand mixtures was increased from 0 to 100%. The modified membrane filter immunofluorescence procedure was used to follow the growth of a strain of chickpea (
Cicer arietinum
)
Rhizobium
in a sterilized oxisol. The results showed a close agreement with viable counts at different stages during the growth cycle. Diluent for the hydrolyzed gelatin also had a marked effect on recovery. The efficiency of release of
Rhizobium
spp. from an oxisol was in the following order for the diluents used: 0.1 M (NH
4
)
2
HPO
4
> 0.1 M Na
2
HPO
4
= 0.1 M sodium-phosphate-buffered saline (pH 7.2) > 0.2 M NH
4
Cl > 0.2 KCl > NaCl = LiCl > water.
Culture medium for selective isolation and enumeration of Gram-negative bacteria from ground meats
Authors: Cyzeska F J , Seiter J A , Marks S N , Jay J M
Published: January 2020

We developed a new medium, designated peptone bile amphotericin cycloheximide (PBAC) agar, which contains (per liter) 10 g of peptone, 300 mg of bile salts, 1 mg of amphotericin B, 1 g of cycloheximide, and 15 g of agar. When 21 samples of fresh ground beef were studied and plate count agar counts were used as references, we obtained a mean recovery of 28% of total counts with violet red bile agar overlay, whereas we obtained 48% recovery with PBAC agar. With 12 samples of frozen ground beef, recovery on violet red bile agar overlay was 29% of the recovery on plate count agar, whereas the corresponding value on PBAC agar was 45%. PBAC agar allowed the enumeration of 1.4 times as many gram-negative bacteria as violet red bile agar overlay. None of eight strains of gram-positive bacteria and none of eight strains of yeasts grew on PBAC agar. Of 158 colonies randomly selected from pour plates of eight fresh ground meat samples, 95% stained gram negative. In comparison, only 70% of 151 colonies selected from corresponding plate count agar plates were gram negative. The lack of background color, turbidity, and ease of use make PBAC agar easier to handle than other media used for gram-negative bacteria, such as violet red bile agar, violet red bile agar overlay, and crystal violet tetrazolium agar. In the preparation PBAC agar, all ingredients are autoclaved together except amphotericin B, which is filter sterilized and added before the plates are poured.
Chemolithotrophic Sulfur-Oxidizing Bacteria from the Galapagos Rift Hydrothermal Vents
Authors: Ruby E. G. , Wirsen C. O. , Jannasch H. W.
Published: January 2020

Three distinct physiological types of sulfur-oxidizing bacteria were enriched and isolated from samples collected at several deep-sea hydrothermal vents (2,550 m) of the Galapagos Rift ocean floor spreading center. Twelve strains of the obligately chemolithotrophic genus
Thiomicrospira
were obtained from venting water and from microbial mats covering surfaces in the immediate vicinity of the vents. From these and other sources two types of obligately heterotrophic sulfur oxidizers were repeatedly isolated that presumably oxidized thiosulfate either to sulfate (acid producing; 9 strains) or to polythionates (base producing; 74 strains). The former were thiobacilli-like, exhibiting a thiosulfate-stimulated increase in growth and CO
2
incorporation, whereas the latter were similar to previously encountered pseudomonad-like heterotrophs. The presence of chemolithotrophic sulfur-oxidizing bacteria in the sulfide-containing hydrothermal water supports the hypothesis that chemosynthesis provides a substantial primary food source for the rich populations of invertebrates found in the immediate vicinity of the vents.
Binding of metals to cell envelopes of Escherichia coli K-12
Authors: Beveridge T J , Koval S F
Published: January 2020

As representative of gram-negative bacteria, the isolated and purified envelopes of an Escherichia coli K-12 strain were used to determine metal-binding capacity. The envelopes were suspended in 5 mM metal solutions for 10 min and 23 degrees C, separated and washed by centrifugation, and analyzed for metal by either atomic absorption or X-ray fluorescence spectroscopy. Of 32 metals tested, large amounts (> 0.9 mumol/mg [dry weight]) of Hf and Os, intermediate amounts (0.1 to 0.4 mumol/mg [dry weight]) of Pb, Zn, Zr, Fe III, Mn, Mo, Mg, Co, and Ce IV, and small amounts (< 0.1 mumol/mg [dry weight]) of Na, K, Rb, Ca, Sr, Cu, Sc, La, Pr, Sm, U, Fe II, Ru, Ni, Hg, Pt, Pd, Au, and In were detected Li and V were not bound to the envelopes. Electron microscopy of unstained, thin-sectioned material provided an electron-scattering profile for localizing the bound metal within the envelope. Energy-dispersive X-ray analysis of thin sections detected all metals in single envelope vesicles. These data suggest that most metal deposition occurred at the polar head group regions of the constituent membranes or along the peptidoglycan layer. No leaching of envelope components was detected by monitoring radioactive probes within the lipopolysaccharide and peptidoglycan layers during metal uptake experiments, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins from metal-loaded envelopes, or protein and carbohydrate determinations on the wash fluids. These results suggest that membrane integrity was not disturbed under these ionic conditions.
Medium for Isolation and Growth of Bacteria Associated with Plum Leaf Scald and Phony Peach Diseases
Authors: Wells J. M. , Raju B. C. , Nyland G. , Lowe S. K.
Published: January 2020

Rickettsia-like bacteria associated with plum leaf scald and phony peach diseases were isolated from diseased but not from healthy tissues and cultured on charcoal-yeast extract medium (BCYE) buffered with ACES (2-[(2-amino-2-oxoethyl) amino]-ethanesulfonic acid). Optimum conditions for isolation and growth on BCYE medium were pH 6.5 to 6.9 at 20 and 25°C under normal atmosphere. Growth of primary colonies and first-passage subcultures was restricted, and colonies reached a maximum diameter of 0.6 mm in 60 days. After 12 passages, subcultures reached maximum growth in 21 days. The rickettsia-like bacteria from BCYE cultures were gram negative, serologically the same as those present in diseased peach and plum, and composed of rod-shaped cells measuring 0.35 by 5 μm (average diameter and maximum length) in a matrix of filamentous strands of similar width but of variable length.
Bacterial adherence to polystyrene: a replica method of screening for bacterial hydrophobicity
Authors: Rosenberg M
Published: January 2020

A simple replica method is described for the rapid identification of colonies of bacteria which adhere to polystyrene. A correlation was found between the adherence of bacterial strains to polystyrene and cell surface hydrophobicity, suggesting the use of this technique in screening for cell surface mutants and in the isolation of hydrophobic bacteria from nature.
Monitoring Steam Sterilization of Surgical Instruments: a Dilemma
Authors: Perkins Robert E. , Bodman H. A. , Kundsin Ruth B. , Walter Carl W.
Published: January 2020

The study of two biological indicators in monitoring “flash” sterilization demonstrated that indicator construction often leads to a false interpretation of spore survival.
Effect of aflatoxin B1 on chromatin-bound ribonucleic acid polymerase and nucleic acid and protein synthesis in germinating maize seeds
Authors: Tripathi R K , Misra R S
Published: January 2020

The treatment of germinating maize seeds (cv. Ganga 2) with aflatoxin B1 resulted in suppression of ribonucleic acid (RNA), protein, and deoxyribonucleic acid (DNA) synthesis at 3, 4, and 5 h, respectively. At or below the concentrations inhibitory for these in vivo syntheses, the toxin inhibited chromatin-bound DNA-dependent RNA polymerase activity. The synthesis of both polyadenylated and non-polyadenylated RNA was inhibited, but the effect on the former was more pronounced. Equilibrium dialysis and difference spectral and viscometric analyses showed a binding of aflatoxin B1 to DNA isolated from the seeds. It is proposed that the inhibition of RNA synthesis in maize seeds by the toxin is due to the interference with the RNA polymerase activity, which seems, at least partially, due to the impairment of DNA template functions.
Enhanced Production of Surfactin from
Bacillus subtilis
by Continuous Product Removal and Metal Cation Additions
Authors: Cooper D. G. , Macdonald C. R. , Duff S. J. B. , Kosaric N.
Published: January 2020

The lipopeptide, surfactin, is produced by
Bacillus subtilis.
A study has been made on large-scale production of this surfactant. A good yield was obtained from a glucose substrate fermentation by continuously removing the product by foam fractionation. The surfactin could be easily recovered from the collapsed foam by acid precipitation. The yield was also improved by the addition of either iron or manganese salts. Hydrocarbon addition to the medium, which normally increases biosurfactant production, completely inhibited surfactin production by
B. subtilis.
Detection of viruses in drinking water by concentration on magnetic iron oxide
Authors: Rao V C , Waghmare S V , Lakhe S B
Published: January 2020

Discharge of raw domestic wastes containing human enteric viruses into water courses, consumption of untreated water from canals, streams, and shallow wells in villages, and cross-contamination of water in the distribution system because of intermittent water supply in urban areas continue to cause widespread outbreaks of infectious hepatitis in India. To detect a low number of viruses in 50- to 100-liter samples of water, a method was developed with magnetic iron oxide as the virus adsorbent. Poliovirus-seeded dechlorinated tap water, adjusted to pH 3.0 and 0.0005 M AlCl3, was filtered through a 10-g bed of iron oxide sandwiched between two AP20 prefilter pads held in a 142-mm-diameter, stainless-steel holder. Virus was eluted from iron oxide by recirculating three times a 100-ml volume of 3% beef extract, pH 9.0. The eluate was reconcentrated to 5 ml by adjusting to pH 3, adding 1 g of iron oxide, stirring for 30 min, and eluting the readsorbed virus with 5 ml of beef extract, pH 9.0. Virus recovery varied from 60 to 80%. Using the above method, we took a survey of drinking water at three locations in Nagpur during 1976 and found the presence of virus in 7 of 50 samples. The quantity of virus recovered ranged from 1 to 7 plaque-forming units per 30 to 60 liters. Virus was detected in some samples even with residual chlorine. No coliforms were detected in the virus-positive samples.