bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Conditioning, Betaine, and Sucrose on Survival of Rhizobacteria in Powder Formulations
Authors: Caesar A. J. , Burr T. J.
Published: January 2020

To assess the feasibility of simplified dry formulations of beneficial rhizobacteria with improved shelf life, strains of
Pseudomonas
and members of the family
Enterobacteriaceae
were conditioned by either aging, exposure to osmotica, or growth on media amended with sucrose or betaine. Strains thus treated were formulated in 1% methylcellulose and talc, and survival was assessed over 10 to 12 months. Survival of 24-h-old cultures of the same strains suspended in 20% xanthan gum and talc over the same interval was used as the standard of comparison. The survival of strains treated with sucrose, with or without betaine, and formulated in methylcellulose and talc was equal to or greater than that of 24-h-old cultures suspended in 20% xanthan gum and talc. Aging of strains on unamended King's medium B, growth of strains on KCl-amended media, and addition of betaine to KCl-amended King's medium B, whether or not strains were suspended in buffer or a diluent isosmotic to culture conditions, failed to improve survival of the strains tested. The practicality of specific amendments, such as sucrose or other disaccharides, for promoting survival of beneficial bacteria in simplified dry formulations was thus demonstrated.
Identification of a new genetic determinant for cell aggregation associated with lactose plasmid transfer in Lactococcus lactis
Authors: van der Lelie D , Chavarri F , Venema G , Gasson M J
Published: January 2020

Derivatives of the lactose miniplasmid pMG820 were constructed in which a staphylococcal erm gene was inserted and in which this was accompanied by subsequent deletion of the lactose genes. The resulting plasmids were thus marked with both erythromycin resistance and lactose utilization genes in pF1132 or solely erythromycin resistance in pF1133. These plasmids retained the normal conjugation properties characteristic of lactose plasmid pLP712, including the generation by intermolecular rearrangement of high-frequency-transfer Clu+ derivatives which exhibited cell aggregation. The use of such Clu+ plasmids in a variety of mating experiments between different lactococcal strains and the observation of cell aggregation when particular mating mixtures were made led to the discovery of a new component of this conjugation system named Agg. A chromosomal gene agg was postulated to be present in some but not all strains of lactococci. High-frequency conjugation and cell aggregation thus depend on the presence of both Agg and Clu, although in a mating pair these components can be in the same or in separate strains. The Agg and Clu components may be analogous to the binding substance and aggregation substance that are involved in the hemolysin plasmid transfer system of Enterococcus faecalis, although control of their expression is different.
Epidemiology of Salmonella sofia in Australia
Authors: Harrington C S , Lanser J A , Manning P A , Murray C J
Published: January 2020

In recent years, the incidence of isolation of Salmonella sofia in Australia has risen from 33% of all poultry isolates in 1982 to a peak of 49% of isolates in 1988. A parallel rise has not been seen in S. sofia isolated from humans. In Israel, however, S. sofia has been commonly isolated from both humans and poultry. We investigated the possibility that the Israeli strains may belong to a different clonal group and express virulence determinants not seen in the Australian isolates, accounting for the apparent differences in the virulence seen within this species. A number of S. sofia isolates from Australian chickens and humans, as well as from Israeli humans and chickens, were compared by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of outer membrane proteins, plasmid profiles, and restriction fragment length polymorphism analysis. No reproducible differences could be detected by analysis of outer membrane proteins. A small 6.4-kb plasmid, pIMVS2, was detected in all Australian isolates from chickens but not in the Israeli isolates. Restriction fragment length polymorphism studies with cosmid clones as probes provided the most discrimination among isolates, allowing us to divide them into seven groups. This technique revealed that significant differences exist between Australian and Israeli isolates and provided additional insights into the epidemiology of these Salmonella isolates.
Succinate transport by a ruminal selenomonad and its regulation by carbohydrate availability and osmotic strength
Authors: Strobel H J , Russell J B
Published: January 2020

Washed cells of strain H18, a newly isolated ruminal selenomonad, decarboxylated succinate 25-fold faster than Selenomonas ruminantium HD4 (130 versus 5 nmol min-1 mg of protein-1, respectively). Batch cultures of strain H18 which were fermenting glucose did not utilize succinate, and glucose-limited continuous cultures were only able to decarboxylate significant amounts of succinate at slow (less than 0.1 h-1) dilution rates. Strain H18 grew more slowly on lactate than glucose (0.2 versus 0.4 h-1, respectively), and more than half of the lactate was initially converted to succinate. Succinate was only utilized after growth on lactate had ceased. Although nonenergized and glucose-energized cells had similar proton motive forces and ATP levels, glucose-energized cells were unable to transport succinate. Transport by nonenergized cells was decreased by small increases in osmotic strength, and it is possible that energy-dependent inhibition of succinate transport was related to changes in cell turgor. Since cells which were deenergized with 2-deoxyglucose or iodoacetate did not transport succinate, it appeared that glycogen metabolism was providing the driving force for succinate uptake. An artificial delta pH drove succinate transport in deenergized cells, but an artificial membrane potential (delta psi) could not serve as a driving force. Because succinate is nearly fully dissociated at pH 7.0 and the transport process was electroneutral, it appeared that succinate was taken up in symport with two protons. An Eadie-Hofstee plot indicated that the rate of uptake was unusually rapid at high substrate concentrations, but the low-velocity, high-affinity component could account for succinate utilization by stationary cultures.(ABSTRACT TRUNCATED AT 250 WORDS)
Intracellular accumulation of potassium and glutamate specifically enhances survival of Escherichia coli in seawater
Authors: Gauthier M J , Flatau G N , Le Rudulier D , Clément R L , Combarro Combarro M P
Published: January 2020

The high resistance of Escherichia coli grown in saline media to seawater was suppressed by an osmotic down-shock. The shock released several molecules into the medium, including potassium, glutamate, and glycine betaine when cells were previously grown in the presence of this osmolyte. Incubation of such sensitized cells in a solution containing K+ (80 mM) and glutamate (50 mM) at pH 7.4 restored their resistance to seawater up to a level close to that observed initially. The protective effect was partly due to the rapid accumulation of K+; a significant exponential relationship between intracellular concentration of K+ and resistance to seawater was observed. Glutamate was accumulated more slowly and progressively completed the action of K+. These data emphasize the specific influence of potassium glutamate on osmotically stressed E. coli cells. They confirm that regulation of osmotic pressure and, probably, of intracellular pH strongly enhances survival of E. coli in seawater. Osmotic fluctuations in waters carrying enteric bacteria from intestines to seawater, together with variations in their K+ and amino acid contents, could modify the ability of cells to survive in marine environments. These results demonstrate the need to strictly control conditions (K+ content, temperature) used to wash cells before their transfer to seawater microcosms. They suggest that the K+ and glutamate contents of media in which E. coli cells are transported to the sea can influence their subsequent survival in marine environments.
Propionic Acid Production by a Propionic Acid-Tolerant Strain of
Propionibacterium acidipropionici
in Batch and Semicontinuous Fermentation
Authors: Woskow Steven A. , Glatz Bonita A.
Published: January 2020

A propionic acid-tolerant derivative of
Propionibacterium acidipropionici
P9 was obtained by serially transferring strain P9 through broth that contained increasing amounts of propionic acid. After 1 year of repeated transfers, a strain (designed P200910) capable of growth at higher propionic acid concentrations than P9 was obtained. An increase in the proportion of cellular straight-chain fatty acids and uncoupling of propionic acid production from growth were observed for strain P200910. Growth rate, sugar utilization, and acid production were monitored during batch and semicontinuous fermentations of semidefined medium and during batch fermentation of whey permeate for both strain P200910 and strain P9. The highest propionic acid concentration (47 g/liter) was produced by P200910 in a semicontinuous fermentation. Strain P200910 produced a higher ratio of propionic acid to acetic acid, utilized sugar more efficiently, and produced more propionic acid per gram of biomass than did its parent in all fermentations.
Biodegradation of dichloromethane and its utilization as a growth substrate under methanogenic conditions
Authors: Freedman D L , Gossett J M
Published: January 2020

Biodegradation of dichloromethane (DCM) to environmentally acceptable products was demonstrated under methanogenic conditions (35 degrees C). When DCM was supplied to enrichment cultures as the sole organic compound at a low enough concentration to avoid inhibition of methanogenesis, the molar ratio of CH4 formed to DCM consumed (0.473) was very close to the amount predicted by stoichiometric conservation of electrons. DCM degradation was also demonstrated when methanogenesis was partially inhibited (with 0.5 to 1.5 mM 2-bromoethanesulfonate or approximately 2 mM DCM) or completely stopped (with 50 to 55.5 mM 2-bromoethanesulfonate). Addition of a eubacterial inhibitor (vancomycin, 100 mg/liter) greatly reduced the rate of DCM degradation. 14CO2 was the principal product of [14C]DCM degradation, followed by 14CH4 (when methanogenesis was uninhibited) or 14CH3COOH (when methanogenesis was partially or completely inhibited). Hydrogen accumulated during DCM degradation and then returned to background levels when DCM was consumed. These results suggested that nonmethanogenic organisms mediated DCM degradation, oxidizing a portion to CO2 and fermenting the remainder to acetate; acetate formation suggested involvement of an acetogen. Methanogens in the enrichment culture then converted the products of DCM degradation to CH4. Aceticlastic methanogens were more easily inhibited by 2-bromoethanesulfonate and DCM than were CO2-reducing methanogens. When DCM was the sole organic-carbon and electron donor source supplied, its use as a growth substrate was demonstrated. The highest observed yield was 0.085 g of suspended organic carbon formed per g of DCM carbon consumed. Approximately 85% of the biomass formed was attributable to the growth of nonmethanogens, and 15% was attributable to methanogens.
Method To Enhance Growth and Sporulation of Pelletized Biocontrol Fungi
Authors: Knudsen G. R. , Eschen D. J. , Dandurand L. M. , Wang Z. G.
Published: January 2020

The biocontrol fungi
Trichoderma harzianum
, used to control soilborne plant pathogens, and
Beauveria bassiana
, used to control insect pests, were formulated as mycelial biomass in alginate pellets with wheat bran added. After drying for 0, 4, or 16 h, pellets were placed in water or in aqueous solutions of polyethylene glycol (PEG) 8000 for 4 to 24 h and then allowed to continue drying. PEG-treated pellets containing
T. harzianum
showed significantly greater proliferation of hyphae in soil than untreated pellets or pellets treated with water. Production of conidia of
T. harzianum
from PEG-treated pellets was lower than production from untreated pellets after 4 days, although rates were equivalent after 7 days. In contrast, production of conidia of
B. bassiana
was significantly more rapid from PEG-treated pellets than from untreated pellets. Biocontrol of soilborne plant pathogens or insect pests may be enhanced by rapid hyphal growth of
T. harzianum
in soil or rapid sporulation of
B. bassiana
on foliage, respectively. Therefore, PEG treatment may improve the efficacy of these biocontrol agents.
Proteases and Their Involvement in the Infection and Immobilization of Nematodes by the Nematophagous Fungus
Arthrobotrys oligospora
Authors: Tunlid Anders , Jansson Sven
Published: January 2020

The nematophagous fungus
Arthrobotrys oligospora
produced extracellular proteases when grown in a liquid culture, as revealed by measuring the hydrolysis of the chromogenic substrate Azocoll. The extracellular protease activity was inhibited by phenylmethylsulfonyl fluoride (PMSF) and other serine protease inhibitors and partly inhibited by the aspartate protease inhibitor pepstatin and by a cysteine protease inhibitor [
l
-
trans
-epoxysuccinyl-leucylamide-(4-guanidino)-butane, or E-64]. Substrate gel electrophoresis showed that the fungus produced several different proteases, including multiple serine proteases. The function of proteases in the infection of nematodes was examined by treating the fungus with various protease inhibitors. None of the inhibitors tested affected the adhesion of nematodes to the traps, but incubating trap-bearing mycelium with a serine protease inhibitor, PMSF, antipain, or chymostatin, or the metalloprotease inhibitor phenanthroline significantly decreased the immobilization of nematodes captured by the fungus. Inhibitors of cysteine or aspartic proteases did not affect the immobilization of captured nematodes. The effects of PMSF on the immobilization of nematodes were probably due to serine proteases produced by the fungus, since the effects were observed when unbound inhibitor was washed away from the fungus before the nematodes were added to the system. No effects were observed when the nematodes only were pretreated with PMSF.
Expression of the Escherichia coli beta-glucuronidase gene in Pseudocercosporella herpotrichoides
Authors: Bunkers G J
Published: January 2020

The plant-pathogenic fungus Pseudocercosporella herpotrichoides has been successfully transformed by using two different positive selection systems in combination with the Escherichia coli gusA gene. The selectable markers used in this study were the hygromycin B phosphotransferase gene (hph) from E. coli and the gene (bml) for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa. A lower transformation rate was obtained with the bml system than with the hph system. Conversely, cotransformation frequencies, as determined with medium plates containing the chromogenic substrate 5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid, were higher with bml than with hph as the selectable marker. The hygromycin-resistant transformants were mitotically stable, and both the selectable gene and gusA were maintained through conidiation. The vector DNA was integrated into the genome, and the number and sites of insertion varied among transformants. Enzyme assays of mycelial extracts showed that beta-glucuronidase activity was highest in transformants with a high gusA copy number. Expression of gusA during growth of the fungus on plants was easily detectable and did not affect pathogenicity. These results form the basis for construction of a versatile and sensitive reporter gene system for P. herpotrichoides.
Aflatoxin production via cross-feeding of pathway intermediates during cofermentation of aflatoxin pathway-blocked Aspergillus parasiticus mutants
Authors: Cleveland T E , Bhatnagar D , Brown R L
Published: January 2020

Cofermentation of Aspergillus parasiticus strains (SRRC 163 and SRRC 2043) blocked at different steps in the aflatoxin B1 (AFB1) biosynthetic pathway in a synthetic liquid medium or on seeds (cottonseed, corn kernels, and peanuts) resulted in production of AFB1. Strain SRRC 2043 accumulated O-methylsterigmatocystin (OMST), a late precursor in AFB1 biosynthesis, whereas SRRC 163 accumulated averantin, an early precursor in the pathway. Strain SRRC 2043 secreted large amounts of OMST in culture relative to the amounts of several other pathway intermediates secreted into media (by other AFB1 pathway-blocked strains). AFB1 production occurred even when colonies of SRRC 163 and SRRC 2043 strains (producing no detectable AFB1) were grown together on an agar medium while physically separated from each other by a filter membrane (0.22-micron pore size). In addition, when mycelia of strain SRRC 163 were added to culture filtrates (containing no mycelia but containing secreted OMST) of strain SRRC 2043, AFB1 production occurred. The results suggested a chemical (rather than genetic) mechanism of complementation for AFB1 production between AFB1 pathway-blocked strains, since no mycelial contact was required between these strains for AFB1 production. The mechanism for chemical complementation involves secretion of OMST by SRRC 2043 and subsequent absorption and conversion of OMST to AFB1 by mycelia of strain SRRC 163.
Detection and production of verotoxin 1 of Escherichia coli O157:H7 in food
Authors: Weeratna R D , Doyle M P
Published: January 2020

Verotoxin 1 (VT1) is a recognized virulence factor of Escherichia coli O157:H7, a cause of severe food-borne disease. The public health significance of preformed verotoxin in food is unknown, and relatively little research has been done to determine the production of VT1 in food. The purposes of this study were to develop a sensitive method to detect VT1 in milk and in ground beef and to determine the conditions for VT1 production in these foods. A sandwich enzyme-linked immunosorbent assay in which we used VT1-specific monoclonal antibody 9C9F5 as the capture antibody and a rabbit polyclonal antibody raised against VT2 as the detection antibody was developed for the detection and quantification of VT1 in milk and in ground beef. The enzyme-linked immunosorbent assay was sensitive to a minimum of 0.5 ng of VT1 per ml of milk and 1.0 ng of VT1 per g of ground beef. The greatest amount of VT1 detected in milk (306 ng/ml) was detected in samples that were incubated at 37 degrees C with agitation (160 rpm) for 48 h. Very little toxin (1 ng/ml) was produced at 25 or 30 degrees C within 96 h. VT1 production was greater in ground beef than in milk; 452 ng of VT1 per g was produced in beef at 37 degrees C in 48 h. Relatively little VT1 was produced in beef within 96 h at 25 and 30 degrees C (2.1 and 9.8 ng of VT1 per g, respectively). Our results indicate that ground beef is a better medium for VT1 production than milk.(ABSTRACT TRUNCATED AT 250 WORDS)
Inhibition of pure cultures of methanogens by benzene ring compounds
Authors: Patel G B , Agnew B J , Dicaire C J
Published: January 2020

The inhibition of methane production by Methanosaeta concilii GP6, Methanospirillum hungatei GP1, Methanobacterium espanolae GP9, and Methanobacterium bryantii M.o.H. during short-term (6-h) exposure to eight benzene ring compounds was studied. The concentration that caused 50% inhibition of the methane production rate (IC50) was dependent on the species and the toxicant. Pentachlorophenol was the most toxic of the tested compounds, with an IC50 of less than 8 mg/liter for all species except M. hungatei. Abietic acid was the next most toxic compound for all the species, with an IC50 in the range of 21.4 to 203 mg/liter. Sodium benzoate was generally the least toxic, with an IC50 in the range of 1,225 to 32,400 mg/liter. 3-Chlorobenzoate was substantially more toxic (IC50, 450 to 1,460 mg/liter) than benzoate. The inhibition by benzene, phenol, vanillic acid, and toluene was intermediate to that of pentachlorophenol and benzoate. Long-term incubation (days) studies to determine effect on growth indicated that all eight compounds were usually much more toxic than predicted from the short-term data. In these latter studies, there was generally a good correlation in the observed inhibition as determined from growth and methane production.
Influence of Potassium in the Agar Medium on the Growth Pattern of the Filamentous Fungus
Fusarium solani
Authors: Das Joachim
Published: January 2020

A decrease in the concentration of K
+
ions below 3 mM in agar medium which also contained starch, casein hydrolysate, MgSO
4
, and K
2
HPO
4
changed the growth pattern of
Fusarium solani
illuminated in diurnal 12-h light/12-h dark cycles from zonation to a feathery growth mode. Rubidium or cesium ions could replace potassium, but lithium, sodium, and the bivalent alkaline earth ions could not.
Phenotypic characterization of CO2-requiring strains of Streptococcus bovis from koalas
Authors: Osawa R , Sly L I
Published: January 2020

We examined phenotypic characteristics of six mannitol-fermenting strains of Streptococcus bovis, including two unusual CO2-requiring strains isolated from koala feces. These strains did not grow in air, but grew in air supplemented with CO2 and under reduced oxygen conditions. All six strains had the same biochemical characteristics, except that the CO2-requiring strains did not produce beta-N-acetylglucosaminidase.
Differences between
Lactobacillus casei
subsp.
casei
2206 and Citrate-Positive
Lactococcus lactis
subsp.
lactis
3022 in the Characteristics of Diacetyl Production
Authors: Kaneko Tsutomu , Watanabe Yukari , Suzuki Hideki
Published: January 2020

Lactobacillus casei
subsp.
casei
2206 exhibited much lower levels of diacetyl reductase activity than Citr
+
Lactococcus lactis
subsp.
lactis
3022 but two-, three-, and more than eightfold-higher levels of diacetyl synthase, lactate dehydrogenase, and NADH oxidase activities, respectively. A requirement for metal ions by the diacetyl synthases in both species was observed. The extracts of strain 2206 but not strain 3022 produced more diacetyl from pyruvate when the reaction for diacetyl synthase was aerated than when it was conducted statically.
Isolation of a neuraminidase gene from Actinomyces viscosus T14V
Authors: Yeung M K , Fernandez S R
Published: January 2020

A genomic library of Actinomyces viscosus T14V DNA in lambda gt11 was screened for expression of neuraminidase activities. Four recombinant clones were detected that gave blue fluorescence upon incubation with a fluorogenic substrate, 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid. Of these, two were identical, and all of the neuraminidase-positive clones shared a common 3.4-kbp DNA region. Expression of the enzyme activities in Escherichia coli carrying the cloned DNA was independent of the lacZ promoter of the vector. Maxicell analysis revealed that the 3.4-kbp DNA insert directed synthesis of a protein with an apparent molecular mass of 100,000 Da. The protein from cell extracts of E. coli clones migrated as a single band that stained for enzyme activity after electrophoresis in a nondissociating polyacrylamide gel. Moreover, human erythrocytes incubated previously with cell lysates from neuraminidase-positive E. coli were hemagglutinated by Actinomyces spp. The enzyme expressed by E. coli was active on substrates containing alpha-2,3 and alpha-2,6 ketosidic linked sialyl residues. Similar substrate specificities were obtained for both the extracellular and cell-associated neuraminidases from A. viscosus T14V. The 3.4-kbp insert hybridized to DNA fragments in a Southern blot containing A. viscosus T14V chromosomal DNA that had been digested with various restriction endonucleases. Data from hybridization studies show that A. viscosus T14V contains a single copy of the neuraminidase gene.
Determinants Encoding Resistance to Several Heavy Metals in Newly Isolated Copper-Resistant Bacteria
Authors: Dressler Cathrin , Kües Ursula , Nies Dietrich H. , Friedrich Bärbel
Published: January 2020

Three copper-resistant, gram-negative bacteria were isolated and characterized. Of the three strains,
Alcaligenes denitrificans
AH tolerated the highest copper concentration (MIC = 4 mM CuSO
4
). All three strains showed various levels of resistance to other metal ions.
A. denitrificans
AH contains sequences which cross-hybridized with the
mer
(mercury resistance) determinant of Tn
21
and the
czc
(cobalt, zinc, and cadmium resistance),
cnr
(cobalt and nickel resistance), and
chr
(chromate resistance) determinants of
A. eutrophus
CH34. DNA-DNA hybridization with probes prepared from
A. eutrophus
CH34 and Tn
21
revealed the presence of
chr-, cnr-
, and
mer
-like sequences on the 200-kb plasmid pHG27 and of
czc, cnr
, and
mer
homologs located on the chromosome. The second strain, classified as
Alcaligenes
sp. strain PW, carries
czc, cnr
, and
mer
homologs on the 240-kb plasmid pHG29-c and a
chr
determinant on the 290-kb plasmid pHG29-a; a third plasmid, the 260-kb large plasmid pHG29-b, is cryptic. In contrast to the
Alcaligenes
strains, which were isolated from metal-contaminated water,
Pseudomonas paucimobilis
CD was isolated from the air. This strain harbors two cryptic plasmids: the 210-kb large plasmid pHG28-a and the 40-kb plasmid pHG28-b. Southern analysis revealed no homology between the metal ion resistance determinants of
A. eutrophus
CH34 and
P. paucimobilis
CD.
Mineralization of the sulfonated azo dye Mordant Yellow 3 by a 6-aminonaphthalene-2-sulfonate-degrading bacterial consortium
Authors: Haug W , Schmidt A , Nörtemann B , Hempel D C , Stolz A , Knackmuss H J
Published: January 2020

Under anaerobic conditions the sulfonated azo dye Mordant Yellow 3 was reduced by the biomass of a bacterial consortium grown aerobically with 6-aminonaphthalene-2-sulfonic acid. Stoichiometric amounts of the aromatic amines 6-aminonaphthalene-2-sulfonate and 5-aminosalicylate were generated and excreted into the medium. After re-aeration of the culture, these amines were mineralized by different members of the bacterial culture. Thus, total degradation of a sulfonated azo dye was achieved by using an alternating anaerobic-aerobic treatment. The ability of the mixed bacterial culture to reduce the azo dye was correlated with the presence of strain BN6, which possessed the ability to oxidize various naphthalenesulfonic acids. It is suggested that strain BN6 has a transport system for naphthalenesulfonic acids which also catalyzes uptake of sulfonated azo dyes. These dyes are then gratuitously reduced in the cytoplasm by unspecific reductases.
Horizontal Variation of Bacterioplankton in the Baltic Sea
Authors: Heinänen Anne , Kuparinen Jorma
Published: January 2020

Variations in the thymidine incorporation rate, bacterial abundance, and mean cell volumes in the surface water (0.5 m) of the Baltic Sea in spring and summer were compared in studies with different spatial scales (570 nautical miles [nmi] [ca. 1056 km], 220 nmi [ca. 407 km], 24 nmi [ca. 44 km], 12 nmi [ca. 22 km], and 200 m). The objective of the comparison was to investigate whether a single sample taken at one sampling point is representative enough for researchers to make generalizations about a larger water area. Bacterioplankton variation was connected more to seasonal characteristics than to the spatial scale of sampling. Variation was greater and more random in spring than in summer. The state variables (bacterial abundance and mean cell volume) varied less than the rate variable (thymidine incorporation). The results suggest that the sampling design for bacterioplankton studies in northern temperate seas should be planned primarily according to the season and that more stress should be put on rate variable measurements than on those of state variables.
Attachment, Chemotaxis, and Multiplication of
Agrobacterium tumefaciens
Biovar 1 and Biovar 3 on Grapevine and Pea
Authors: Brisset Marie-Noëlle , Rodriguez-Palenzuela Pablo , Burr Thomas J. , Collmer Alan
Published: January 2020

Tumorigenic (CG49) and nontumorigenic (CG484) strains of
Agrobacterium tumefaciens
bv. 3 attached to grape roots at a higher level than did a nonpectinolytic mutant of CG49 (CG50) or a tumorigenic strain of
A. tumefaciens
bv. 1 (CG628). Strains attached equally well to wounded and unwounded grape roots. Strains responded differently to pea plants in that biovar 3 strains consistently attached to unwounded roots at a lower level than they did to wounded roots, whereas CG628 attached equally well regardless of wounding. The lowest levels of attachment to pea roots were consistently observed for CG50. Population curves were calculated for the strains inoculated into wound sites on grape and pea roots.
A. tumefaciens
bv. 3 wild-type strains developed greater populations at wound sites on grape roots after 100 h (resulting in root decay) than did CG50 or CG628. Population curves for strains at wound sites on pea roots were different from those on grape roots. There were no significant differences in populations after 100 h, and no strains caused root decay. No differences in the chemotaxis of wild-type and mutant
A. tumefaciens
bv. 3 strains towards grape roots, crown pieces, or root extracts were observed, but the biovar 1 strain, CG628, always migrated the greatest distance towards all substrates. Polygalacturonase production may affect attachment to grape roots and multiplication of
A. tumefaciens
bv. 3 at wound sites and thus be associated with the specificity of the bacterium for grape.
Role of hydrosulfide ions (HS-) in methylmercury resistance in Saccharomyces cerevisiae
Authors: Ono B , Ishii N , Fujino S , Aoyama I
Published: January 2020

Methylmercury-resistant mutants were obtained from Saccharomyces cerevisiae. They were divided into two complementation groups, met2 (homoserine O-acetyltransferase deficiency) and met15 (enzyme deficiency unknown), as reported previously. It was found that met15 was allelic to met17 (O-acetylserine and O-acetylhomoserine sulfhydrylase deficiency). Methylmercury toxicity was counteracted by exogenously added HS-, and both met2 and met17 (met15) mutants overproduced H2S. On the basis of these results, we conclude that met2 and met17 (met15) cause accumulation of hydrosulfide ions in the cell and that the increased level of hydrosulfide is responsible for detoxification of methylmercury.
Hydrophobicity, Adhesion, and Surface-Exposed Proteins of Gliding Bacteria
Authors: Sorongon Maria L. , Bloodgood Robert A. , Burchard Robert P.
Published: January 2020

The cell surface hydrophobicities of a variety of aquatic and terrestrial gliding bacteria were measured by an assay of bacterial adherence to hydrocarbons (BATH), hydrophobic interaction chromatography, and the salt aggregation test. The bacteria demonstrated a broad range of hydrophobicities. Results among the three hydrophobicity assays performed on very hydrophilic strains were quite consistent. Bacterial adhesion to glass did not correlate with any particular measure of surface hydrophobicity. Several adhesion-defective mutants of
Cytophaga
sp. strain U67 were found to be more hydrophilic than the wild type, particularly by the BATH assay and hydrophobic interaction chromatography. The very limited adhesion of these mutants correlated well with hydrophilicity as determined by the BATH assay. The hydrophobicities of several adhesion-competent revertants ranged between those of the wild type and the mutants. As measured by the BATH assay, starvation increased hydrophobicity of both the wild type and an adhesion-defective mutant. During filament fragmentation of
Flexibacter
sp. strain FS-1, marked changes in hydrophobicity and adhesion were accompanied by changes in the arrays of surface-exposed proteins as detected by an immobilized radioiodination procedure.
Oxygen Uptake and Hydrogen-Stimulated Nitrogenase Activity from
Azorhizobium caulinodans
ORS571 Grown in a Succinate-Limited Chemostat
Authors: Allen George C. , Grimm Daniel T. , Elkan Gerald H.
Published: January 2020

Succinate-limited continuous cultures of an
Azorhizobium caulinodans
strain were grown on ammonia or nitrogen gas as a nitrogen source. Ammonia-grown cells became oxygen limited at 1.7 μM dissolved oxygen, whereas nitrogen-fixing cells remained succinate limited even at dissolved oxygen concentrations as low as 0.9 μM. Nitrogen-fixing cells tolerated dissolved oxygen concentrations as high as 41 μM. Succinate-dependent oxygen uptake rates of cells from the different steady states ranged from 178 to 236 nmol min
−1
mg of protein
−1
and were not affected by varying chemostat-dissolved oxygen concentration or nitrogen source. When equimolar concentrations of succinate and β-hydroxybutyrate were combined, oxygen uptake rates were greater than when either substrate was used alone. Azide could also used alone as a respiratory substrate regardless of nitrogen source; however, when azide was added following succinate additions, oxygen uptake was inhibited in ammonia-grown cells and stimulated in nitrogen-fixing cells. Use of 25 mM succinate in the chemostat resevoir at a dilution rate of 0.1 h
−1
resulted in high levels of background respiration and nitrogenase activity, indicating that the cells were not energy limited. Lowering the reservoir succinate to 5 mM imposed energy limitation. Maximum succinate-dependent nitrogenase activity was 1,741 nmol of C
2
H
4
h
−1
mg (dry weight)
−1
, and maximum hydrogen-dependent nitrogenase activity was 949 nmol of C
2
H
4
h
−1
mg (dry weight)
−1
. However, when concentration of 5% (vol/vol) hydrogen or greater were combined with succinate, nitrogenase activity decreased by 35% in comparison to when succinate was used alone. Substitution of argon for nitrogen in the chemostat inflow gas resulted in “washout,” proving that ORS571 can grow on N
2
and that there was not a nitrogen source in the medium that could substitute.
Propidium iodide as an indicator of Giardia cyst viability
Authors: Sauch J F , Flanigan D , Galvin M L , Berman D , Jakubowski W
Published: January 2020

The use of propidium iodide, whose uptake indicates cell death or damage, was investigated to assess the viability of heat-inactivated and chemically inactivated Giardia muris cysts. This was done by comparing propidium iodide staining with excystation. We first determined that propidium iodide could be used with an immunofluorescence detection procedure by showing that the percentages of Giardia lamblia cysts stained with this dye before and after subjecting them to a fluorescence detection method were similar. G. muris cysts were then exposed to heat (56 degrees C), 0.5 to 4 mg of chlorine per liter (pH 7.0, 5 degrees C), 0.1 to 10 mg of a quaternary ammonium compound per liter, or 2 mg of preformed and forming monochloramine per liter (pH 7.2, 18 to 20 degrees C). A good positive correlation between percent propidium iodide-stained cysts and lack of excystation was demonstrated for G. muris cysts exposed either to heat or to the quaternary ammonium compound. However, no significant correlation between absence of excystation and propidium iodide staining was found for cysts exposed to chlorine or monochloramines. These results demonstrate that the propidium iodide staining procedure is not satisfactory for determining the viability of G. muris cysts exposed to these two commonly used drinking water disinfectants.
Frankia genus-specific characterization by polymerase chain reaction
Authors: Simonet P , Grosjean M C , Misra A K , Nazaret S , Cournoyer B , Normand P
Published: January 2020

The polymerase chain reaction (PCR) is an in vitro procedure for primer-directed enzymatic amplification of specific template nucleic acid sequences. In order to determine whether a given actinomycete isolated from an actinorhiza (nodule) belongs to the genus Frankia or is a contaminant, we have developed a test based on the PCR. Primers complementary to sequences of two DNA regions corresponding to the nif genes (nifH and nifD) and the rRNA genes (16S and 23S) were specifically chosen to differentially amplify DNAs from Frankia strains but not those from other microorganisms. A series of positive and negative controls were set up by using universal or selective primers resulting in a discriminant amplification, which could be detected after agarose gel electrophoresis. In the nif region, degenerate oligonucleotide primers were used to amplify a target common to all the nitrogen-fixing microorganisms tested, while another set of primers amplified a target with a high specificity for Frankia strains. In the rRNA gene region, universal and specific primers were characterized and tested with DNAs from a wide range of microorganisms. The efficiency of this rapid and sensitive PCR assay was tested with an isolate obtained from Alnus nepalensis nodules, confirming results obtained by nodulation tests.
High-Level Nickel Resistance in
Alcaligenes xylosoxydans
31A and
Alcaligenes eutrophus
KTO2
Authors: Schmidt Thomas , Stoppel Ralf-D. , Schlegel Hans G.
Published: January 2020

Two new nickel-resistant strains of
Alcaligenes
species were selected from a large number (about 400) of strains isolated from ecosystems polluted by heavy metals and were studied on the physiological and molecular level.
Alcaligenes xylosoxydans
31A is a heterotrophic bacterium, and
Alcaligenes eutrophus
KTO2 is an autotrophic aerobic hydrogen-oxidizing bacterium. Both strains carry—among other plasmids—a megaplasmid determining resistance to 20 to 50 mM NiCl
2
and 20 mM CoCl
2
(when growing in defined Tris-buffered media). Megaplasmids pTOM8, pTOM9 from strain 31A, and pGOE2 from strain KTO2 confer nickel resistance to the same degree to transconjugants of all strains of
A. eutrophus
tested but were not transferred to
Escherichia coli.
However, DNA fragments carrying the nickel resistance genes, cloned into broad-hostrange vector pVDZ'2, confer resistance to
A. eutrophus
derivatives as well as
E. coli.
The DNA fragments of both bacteria, TBA8, TBA9, and GBA (14.5-kb
Bam
HI fragments), appear to be identical. They share equal size, restriction maps, and strong DNA homology but are largely different from fragment HKI of nickel-cobalt resistance plasmid pMOL28 of
A. eutrophus
CH34.
Two novel strains of Bacillus thuringiensis toxic to coleopterans
Authors: Rupar M J , Donovan W P , Groat R G , Slaney A C , Mattison J W , Johnson T B , Charles J F , Dumanoir V C , de Barjac H
Published: January 2020

Two novel strains of Bacillus thuringiensis were isolated from native habitats by the use of genes coding for proteins toxic to coleopterans (cryIII genes) as hybridization probes. Strain EG2838 (isolated by the use of the cryIIIA probe) contained a cryIIIA-hybridizing plasmid of approximately 100 MDa and synthesized crystal proteins of approximately 200 (doublet), 74, 70, 32, and 28 kDa. Strain EG4961 (isolated by the use of a cryIIIA-related probe) contained a cryIIIA-hybridizing plasmid of approximately 95 MDa and synthesized crystal proteins of 74, 70, and 30 kDa. Structural relationships among the crystal proteins of strains EG2838 and EG4961 were detected; antibodies to the CryIIIA protein toxic to coleopterans reacted with the 74- and 70-kDa proteins of EG2838 and EG4961, antibodies to the 32-kDa plus 28-kDa proteins of EG2838 reacted with the 30-kDa protein of EG4961, and antibodies to the 200-kDa proteins of EG2838 reacted with the 28-kDa protein of EG2838. Experiments with B. thuringiensis flagella antibody reagents demonstrated that EG2838 belongs to H serotype 9 (reference strain B. thuringiensis subsp. tolworthi) and that EG4961 belongs to H serotype 18 (reference strain B. thuringiensis subsp. kumamotoensis). A mixture of spores plus crystal proteins of either EG2838 or EG4961 was toxic to the larvae of Colorado potato beetle (Leptinotarsa decemlineata), and significantly, the EG4961 mixture was also toxic to the larvae of southern corn rootworm (Diabrotica undecimpunctata howardi). DNA restriction blot analysis suggested that strains EG2838 and EG4961 each contained a unique gene coding for a protein toxic to coleopterans.
Effect of the lactoperoxidase system on Listeria monocytogenes behavior in raw milk at refrigeration temperatures
Authors: Gaya P , Medina M , Nuñez M
Published: January 2020

Activity of raw milk lactoperoxidase-thiocyanate-hydrogen peroxide (LP) system on four Listeria monocytogenes strains at refrigeration temperatures after addition of 0.25 mM sodium thiocyanate and 0.25 mM hydrogen peroxide was studied. The LP system exhibited a bactericidal activity against L. monocytogenes at 4 and 8 degrees C; the activity was dependent on temperature, length of incubation, and strain of L. monocytogenes tested. D values in activated-LP system milk for the four strains tested ranged from 4.1 to 11.2 days at 4 degrees C and from 4.4 to 9.7 days at 8 degrees C. The lactoperoxidase level in raw milk declined during a 7-day incubation, the decrease being more pronounced at 8 degrees C than at 4 degrees C and in control milk than in activated-LP system milk. The thiocyanate concentration decreased considerably in activated-LP system milk at both temperatures during the first 8 h of incubation. LP system activation was shown to be a feasible procedure for controlling development of L. monocytogenes in raw milk at refrigeration temperatures.
Effects of farnesol and the off-flavor derivative geosmin on Streptomyces tendae
Authors: Dionigi C P , Millie D F , Johnsen P B
Published: January 2020

Effects of the sesquiterpene farnesol (3,7,11-trimethyl-2,6,10-dodecatrien-1-ol) and the sesquiterpene derivative geosmin (1,10-trans-dimethyl-trans-9-decalol) were investigated in a geosmin-producing actinomycete, Streptomyces tendae. Exposure to 300 microM farnesol reduced biomass (fresh matter) accumulation by 97% compared with biomass accumulation by controls, whereas an equal amount of geosmin did not affect biomass accumulation. Increasing exposure to farnesol corresponded with reduced optical density of the culture, reduced levels of geosmin, and reduced metabolic heat production compared with controls, while exogenous geosmin did not affect these parameters. Geosmin dissipated from unioculated medium more rapidly than farnesol, indicating that in addition to the lower toxicity of geosmin, the actual exposure to geosmin over time may be less than exposure to an equal amount of farnesol. Cultures grown on Actinomyces-B medium contained 99.5% less geosmin and were more sensitive to farnesol than those grown on Hickey-Tresner medium, indicating that geosmin synthesis was associated with reduced sensitivity to farnesol. Consumption of farnesyl moieties during geosmin synthesis may reduce the potential for farnesol-induced inhibition of growth and metabolism.
Screening of Natural Waters for Viruses Which Infect
Chlorella
Cells
Authors: Yamada Takashi , Higashiyama Takanobu , Fukuda Takao
Published: January 2020

By using a plaque assay with the unicellular green alga
Chlorella
sp. strain NC64A as a host, viruses were screened from natural pond waters collected in Kyoto and Higashi-Hiroshima, Japan. From some samples tested, two kinds of plaques, large (φ = 6 to 10 mm) and small (φ = 2 to 3 mm), were detected with various frequencies. The frequency of plaques in each of the water sources was seasonal; generally, it reached a peak value (8,000 PFU/ml) in May and gradually decreased to the limit of detection (<1) in November before increasing again in early spring. Electron microscopy revealed that the purified and negatively stained viruses were very large (125 to 200 nm) icosahedral particles. The genome isolated from these particles was always a linear double-stranded DNA of 340 to 370 kbp. Electrophoresis patterns of the DNA fragments produced by digestion with restriction enzymes differed considerably from plaque to plaque, even for plaques from the same water source. However, Southern hybridization showed strong homology among all of the virus DNAs tested, indicating relatedness of those viruses. A possible use of the
Chlorella
virus assay system to monitor the natural population of algal cells and water quality is discussed.
Serogrouping of Halophilic Bdellovibrios from Chesapeake Bay and Environs by Immunodiffusion and Immunoelectrophoresis
Authors: Schoeffield Andrew J. , Falkler William A. , Desai Darshana , Williams Henry N.
Published: January 2020

Little has been reported on the serological relationship of halophilic bdellovibrios (Bd). Immunodiffusion analysis performed with rabbit or mouse Bd antisera developed against eight halophilic Bd isolates and one terrestrial Bd isolate, when reacted with soluble antigen preparations of 45 isolates of halophilic Bd, allowed separation into seven serogroups, which were distinct from the terrestrial isolate. Soluble antigen preparations of prey bacteria,
Vibrio parahaemolyticus
P-5 (P-5) and
Escherichia coli
ML 35 (ML 35), exhibited no reactivity with the antisera by immunodiffusion. Immunoelectrophoresis revealed the presence of three distinct antigens in homologous reactions and one shared antigen in heterologous Bd reactions. Shared antigens were noted between halophilic and terrestrial Bd, in addition to between halophilic Bd strains, indicating the possible existence of an antigen(s) which may be shared among all Bd. Again, no shared antigen was noted when P-5 or ML 35 was allowed by immunoelectrophoresis to react with the antisera. Prey susceptibility testing of the seven distinct groups of halophilic Bd, using 20 test prey, produced essentially identical spectra for each group, indicating that this was not a useful technique in delineating the Bd. While immunoelectrophoresis was able to demonstrate an antigen common to all Bd tested, immunodiffusion was able to delineate strains on the basis of a “serogroup specific” antigen. This suggests that immunological tools may serve as important means to study the taxonomy of halophilic Bd, as well as in the formation of a clearer taxonomic picture of the genus
Bdellovibrio.
Detection of Plasmid Transfer from
Pseudomonas fluorescens
to Indigenous Bacteria in Soil by Using Bacteriophage φR2f for Donor Counterselection
Authors: Smit Eric , van Elsas Jan D. , van Veen Johannes A. , de Vos Willem M.
Published: January 2020

The transfer of a genetically marked derivative of plasmid RP4, RP4p, from
Pseudomonas fluorescens
to members of the indigenous microflora of the wheat rhizosphere was studied by using a bacteriophage that specifically lyses the donor strain and a specific eukaryotic marker on the plasmid. Transfer of RP4p to the wheat rhizosphere microflora was observed, and the number of transconjugants detected was approximately 10
3
transconjugants per g of soil when 10
7
donor cells per g of soil were added; transfer in the corresponding bulk soil was slightly above the limit of detection. All of the indigenous transconjugants which we analyzed contained a 60-kb plasmid and were able to transfer this plasmid to a Nx
r
Rp
r
P. fluorescens
recipient strain. The indigenous transconjugants were identified as belonging to
Pseudomonas
spp.,
Enterobacter
spp.,
Comamonas
spp., and
Alcaligenes
spp.
Effects of solvents and alcohols on the polar lipid composition of Clostridium butyricum under conditions of controlled lipid chain composition
Authors: MacDonald D L , Goldfine H
Published: January 2020

Clostridium butyricum has been grown in media devoid of biotin, to which long-chain fatty acids have been added to promote growth. We have shown previously that, under these conditions, exogenous fatty acids are extensively incorporated into the cellular phospholipids. Cells grown with elaidic acid, trans-9-18:1, have normal ratios of the glycerol acetal of plasmenylethanolamine (GAPlaE) to phosphatidylethanolamine (PE) plus plasmenylethanolamine (PlaE) compared with cells grown with biotin. When ethanol, cyclohexane, or n-octanol was added to elaidate-containing media, the ratio of GAPlaE to PE plus PlaE was significantly increased. Addition of dodecane and n-butanol did not affect this ratio. When cells were grown with oleic acid in the absence of biotin, the GAPlaE to PE plus PlaE ratio was increased 5.4-fold compared with elaidate-grown cells. In oleate-supplemented media, the addition of solvents or n-alcohols produced no further increase in this ratio. We conclude that these changes in lipid composition represent cellular responses to perturbation of the equilibria between the lamellar and nonlamellar liquid crystalline phases in the cell membrane.