Studies on histoplasmosis I. Comparative virulence of variant and parent strain Histoplasma capsulatum in Hamsters

Summary 1. A variant strain ofHistoplasma capsulatum was isolated from a secondary colony in cultures of tissue from hamsters infected with the parent strain ofHistoplasma. The variant strain was found to differ from the parent strain in colonial morphology, stability of the yeast phase in subculture, and in growth rate.2. Results of virulence testing with these two strains showed the parent strain to be somewhat more virulent than the variant strain as measured by survival time and slope of the death rate curve over a 19 day period.3. Nevertheless, in view of the more rapid growth rate of the variant and its probable antigenic variation from the parent strain, the development of such a variant is viewed as an increased hazard to the host, particularly in a longer term infection.4. The repeated isolation of a variant strain ofHistoplasma suggests that strain variation may play a significant role in the outcome of naturally acquired infections.This work was initiated at the State University of New York, Downstate Medical Center, Brooklyn, N. Y., and supported in part by funds ((#12-1081) from the National Fund for Medical Education, grant # 12-1054.     

Selection of a Photosynthetic Bacterium Suitable for Hydrogen Production in Outdoor Cultures among Strains Isolated in the Seoul,Taegu, Sendai and Bangkok Areas

Very efficient hydrogen producing photosynthetic bacteria, strains SL1, SL3, SL16 and TG28 newly isolated in Korea, and strain KM113 newly isolated in the Sendai area, were found to be Rhodopseudomonas spp. To examine the stability of cell suspensions of the cultures for hydrogen production, which is closely associated with light absorption, we conducted larger scale cultures under periodic illumination (12-hr intervals) without stirring at 30°C using strains SL1 and Rhodopseudomonas sphaeroides B5, the latter was isolated in the Bangkok area. Both strains gave homogeneous cell suspensions throughout the incubation period and larger amounts of hydrogen were produced in a shorter period of time by both cultures than obtained with Rhodopseudomonas sp. TN3, an isolate from the Sendai area which was reported previously. With the cells of the new isolates and strains TN3 and B5 grown on glutamate-malate medium at 30°C, we measured hydrogen production at 20, 30 and 40°C in the same medium. Among them, strains SL1, SL16 and KM113 showed the highest hydrogen production activity at 30°C. The maximum hydrogen production rates with these strains were over 130 µ1/hr/mg dry cells, but at 40°C, the highest activity (138 µl/hr/mg dry cells) was obtained with strain B5. Since strain B5 also showed good activities at 20 and 30°C, we suggest that this strain might be suitable for hydrogen production in outdoor cultures.     

Haplopappus gracilis cell strains resistant to pyrimidine analogues

Summary Strains of Haplopappus gracilis (Nutt.) Gray cells resistant to 6-azauracil have been isolated from cultures of diploid cells. These strains are also resistant to 8-azaguanine, as is their parent. The variants are 100- to 125-fold more resistant to 6-azauracil than their parent, and they exhibit different spectra of cross resistance to other pyrimidine analogues. The phenotype of each variant is stable in the absence of selection. The majority of cells in cultures of the variants are diploid; all others examined were tetraploid. Initial rates of uptake of uracil are not reduced in the variants. Fluorouracil, to which two variants are resistant, is taken up by one of them as well as by the parent. Responses of the other two to fluorouracil are not correlated with decreased ability to accumulate this analogue.     

GROWTH INDUCTION IN CULTURES OF HAPLOPAPPUS GRACILIS. II. THE BEHAVIOR OF THE NUCLEUS

Mitra , J., and F. C. Steward . (Cornell U., Ithaca, New York.) Growth induction in cultures of Haplopappus gracilis. II. The behavior of the nucleus. Amer. Jour. Bot. 48(5): 358–368. Illus. 1961.—Cells of Haplopappus, which have been stimulated to grow under the influence of coconut milk and such synergists as naphthalene or 2,4-dichlorophenoxy acetic acid, can be cultivated as free cells, as small cell clusters, or as peripheral cells on a cultured colony or mass. Such cells display forms and cell lineages, the general pattern of which is reminiscent of those that may occur in early embryogeny. To this extent, Haplopappus resembles what has previously been observed in the growth of free cells of carrot. The form of the normal chromosome of Haplopappus (2n = 4) is described with respect to root tip cells. The range of effects which may be observed in the nuclei of the cultured cell is also described. Such variations as the following were encountered: (1) multinucleate giant cells which may divide by internal segmentation; (2) polyploidy, giving rise to highly polyploid nuclei up to, and even in excess of, 64 chromosomes; (3) somatic pairing; (4) haploidy; (5) pseudochiasmata, with the evident implication of somatic “crossing-over”; (6) chromosome breaks, reunions and bridges, such as are commonly associated with effects of radiation and with chemical mutagens. Attention is drawn to the usefulness of this material for the further experimental investigation of the biochemical basis for the cytological events which are here described, and, if the variant cells may be cloned, of the relationship between the aberrant nuclear cytology and the ability of the cells or colonies to differentiate or to undergo morphogenesis.     

Preliminary characterization of an ineffective Frankia derived from a spontaneously neomycin-resistant strain

Five strains of Frankia isolated from actinorhizae of 3 alder species were cultivated with various concentrations of neomycin in the medium. For one strain, resistance to neomycin was associated with lost of effectiveness. The ineffective strain was as infective as the parent strain. Protein and isoenzymes patterns showed that the ineffective and the parent strains were quite similar. The ineffective strain differentiated vesicles inside the infected host-cells.     

Mutations in the rfbT Gene Are Responsible for the Ogawa to Inaba Serotype Conversion in Vibrio cholerae O1

In cultures of Vibrio cholerae strains of Ogawa serotype, variant strains which had undergone serotype conversion from Ogawa to Inaba were identified. The rfbT genes cloned from the parent strains were found to produce a 31-kDa protein in the maxicell system, and to cause serotype conversion when introduced into E. coli cells expressing Inaba serotype specificity. On the other hand, rfbT genes cloned from the variant strains neither produced the 31-kDa protein nor caused serotype conversion. Nucleotide sequence of these rfbT genes as well as those of two clinical Vibrio cholerae strains of Inaba serotype revealed that mutations causing premature termination of their rfbT genes were invariably present in strains expressing Inaba serotype specificity. The result strongly suggested that genetic alteration of the rfbT gene is responsible for serotype conversion of Vibrio cholerae O1.     

Characteristics of Group A Streptococcal Variants Treated with Mitomycin C

Transfers of Streptococcus pyogenes strain T12 in Todd–Hewitt broth containing stepwise increases in amounts of mitomycin C (MC) gave rise to slight changes of their colonial appearances. Variants thus obtained were examined for antibiotic and bile resistances; production of streptolysin-S, -O and deoxyribonuclease; growth in alkaline medium, high salt concentration, and at 10 C and 45 C; sugar fermentations, and precipitin reactions. Four strains retained group A antigen, but some of them lost the ability to produce hemolysins and deoxyribonuclease, and acquired resistance to bile, penicillin and streptomycin as well as MC, and to physical environments. Four other strains lost group A antigen and acquired new antigens common to cells of group C, group D, or highly antibiotic-resistant mutants reported previously. A variant which reacted with group C antiserum contained galactosamine, but not glucosamine, while the parent strain showed the reverse pattern. Many other variants contained both hexosamines. Even a variant, strain TL3-2, reacted strongly only with group A antiserum, but contained glucose and both hexosamines. These strains having galactosamine possessed uridine diphosphate (UDP)-N-acetylglucosamine-4-epimerase activity which converted the substrate into UDP-N-acetylgalactosamine, while the parent strain failed to demonstrate the existence of this enzyme. The variants were discussed with respect to the group A streptococcal variations possessing no more original characteristics.     

Characterization and infectivity of a spontaneous variant isolated fromFrankia sp. WEY 0131391

Summary A spontaneous variant, obtained from aFrankia isolate fromAlnus rubra nodules, was compared with the parent strain with regard to infectivity, nitrogenase activity, and electrophoretic and immunological profiles. Both the parent and the variant strain were equally effective in inducing nodulation in seedlings ofA. rubra. All inoculated plants had an active nitrogenase system as measured by the acetylene reduction assay. Electrophoresis of whole cell homogenates on SDS-polyacrylamide slab gels showed similar electrophoretic profiles; however, the variant strain also exhibited striking differences in protein patterns that distinguish it from the parent strain. Immunological analysis of the originalFrankia strain and its variant revealed shared antigens as well as immunologically distinct antigenic determinants in the two strains. The variant strain exhibits a distinct morphology and growth patterns which remain stable after many passages through culture.     

Formation of cellulose fibrils by gram-negative bacteria and their role in bacterial flocculation

Summary Exocellular fibrils, consisting of true cellulose, were found to be produced by many bacteria. These bacteria have been selected out of a large number of strains isolated from activated sludge on the basis of their flocculent growth habit in liquid medium.The amount of cellulose, present in the bacterial flocs, varied from 1.0 to 4.0%. In addition to strains isolated from activated sludge, like Pseudomonas, Achromobacter, Alcaligenes and Aerobacter, also strains of the genera Rhizobium, Agrobacterium and Azotobacter were found to give flocculent growth due to the formation of cellulose fibrils.Bacterial flocculation in pure cultures of the strains examined was mainly caused by the production of exocellular fibrils. Apparently, the formation of cellulose fibrils seems to be a common property of Gram-negative, floc-forming bacteria, and may not be restricted to Acetobacter xylinum.     

Effect of citrate on growth of Lactococcus lactis subsp. lactis in milk

The effect of citrate on the growth of Lactococcus lactis subsp. lactis var. diacetylactis in milk has been investigated. Five strains of Lactococcus lactis subsp. lactis var. diacetylactis were compared to their citrate-negative variants, which lack the plasmid coding for citrate permease. In most cases, acidification kinetics and the final bacterial concentration of pure cultures of parental and variant strains did not differ significantly. Co-cultures of parental and variant strains, however, systematically tended towards the predominance of parental strains. Citrate metabolism is responsible for this change, since the predominance of citrate-positive strains was not observed in the absence of citrate. Continuous culture in milk enabled the difference in growth rates between the parental strain Lactococcus lactis subsp. lactis var. diacetylactis CDI1 and its citrate-negative variant to be quantified by following changes in the populations of the two co-cultured strains. At 26 °C, the growth rate of the parental strain was 7% higher than that of its citrate-negative variant. These results show that citrate metabolism slightly stimulates the growth of lactococci in milk. Received: 18 February 1997 / Received revision: 2 May 1997 / Accepted: 4 May 1997     

Resistance system of Mycobacterium bovis B.C.B. to aminoglycoside- and peptide-antibiotics. Comparison of resistant phenotypes between Mycobacterium tuberculosis and Mycobacterium bovis

The resistance system of Mycobacterium bovis (B.C.G.) to aminoglycoside-and peptide-antibiotics has been studied. The phenotype of mutants isolated from the parent B.C.G. strain by a single-step selection with an antibiotic were classified into the following three types: (1) resistant only to a low concentration (200 μg/ml) of kanamycin in Ogawa egg medium (k1R); (2) resistant to a low concentration (200 μg/ml) of viomycin and of capreomycin (2R); and (3) resistant to a high concentration (1,000 μg/ml or more) of kanamycin and low concentrations (100 to 200 μg/ml) of lividomycin and of paromomycin (KR). The mutants showing these phenotypes, k1R, 2R, and KR, were isolated from the parent strain by inoculating the strain into media containing 100 μg/ml of kanamycin, and 100 μ/g/ml of viomycin or capreomycin, and 1,000 μg/ml of kanamycin, respectively, at rates of 10^?5-10^?6, 10^?5-10^?6, and 10^?6-10^?7, respectively, in a total viable population of the parent strain. Unlike in the case of M. tuberculosis, no mutant could be isolated from the parent strain by use of enviomycin, lividomycin, and/or paromomycin. In contrast to the fact that quadruply resistant mutants were isolated directly from the parent H37Rv strain of M. tuberculosis, such mutants could be isolated only by two-step selections. Furthermore, the phenotypes of the quadruply resistant mutants were those showing a higher resistance or a broader spectrum than expected by the addition of phenotypes of individual mutations. In addition, it was shown that, in contrast to the fact that hextuply resistant mutants could be isolated directly from the parent strain of M. tuberculosis, such mutants were not isolated directly from the parent B.C.G. strain, but could be isolated only after pre-incubation of the strain on a medium containing Tween 80.     

Isolation of a mutant strain of Pseudomonas sp ATCC 31461 exhibiting elevated polysaccharide production

A mutant strain of the bacterium Pseudomonas sp. ATCC 31461 that exhibited elevated production of the polysaccharide gellan on glucose or corn syrup as a carbon source was isolated. Gellan production by the mutant strain was about twofold higher than its parent strain on glucose or corn syrup after 48 h of growth, and about 1.4-fold higher after 72 h. An increase in biomass production was not correlated with enhanced gellan synthesis by the mutant strain. The increased gellan production by the mutant strain on either carbon source resulted in an increase in its culture medium viscosity and the viscosity of the isolated polysaccharide produced by glucose-grown cells. No differences in the glucuronic acid content of the polysaccharides produced by the mutant and parent strains were observed. Journal of Industrial Microbiology & Biotechnology (2002) 29, 185–188 doi:10.1038/sj.jim.7000278 Received 13 February 2002/ Accepted in revised form 20 May 2002     

Genetic variability in <Emphasis Type="Italic">Bradyrhizobium japonicum</Emphasis> strains nodulating soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merrill]

Brazil has succeeded in sustaining production of soybean [Glycine max (L.) Merrill] by relying mainly on symbiotic N₂ fixation, thanks to the selection and use in inoculants of very effective strains of Bradyrhizobium japonicum and Bradyrhizobium elkanii. It is desirable that rhizobial strains used in inoculants have stable genetic and physiological traits, but experience confirms that rhizobial strains nodulating soybean often lose competitiveness in the field. In this study, soybean cultivar BR 16 was single-inoculated with four B. japonicum strains (CIAT 88, CIAT 89, CIAT 104 and CIAT 105) under aseptic conditions. Forty colonies were isolated from nodules produced by each strain. The progenitor strains, the isolates and four other commercially recommended strains were applied separately to the same cultivar under controlled greenhouse conditions. We observed significant variability in nodulation, shoot dry weight, shoot total N, nodule efficiency (total N mass over nodule mass) and BOX-PCR fingerprinting profiles between variant and progenitor strains. Some variant strains resulted in significantly larger responses in terms of shoot total N, dry weight and nodule efficiency, when compared to their progenitor strain. These results highlight the need for intermittent evaluation of stock bacterial cultures to guarantee effective symbiosis after inoculation. Most importantly, it indicates that it is possible to improve symbiotic effectiveness by screening rhizobial strains for higher N₂ fixation capacity within the natural variability that can be found within each progenitor strain.     

Antigens of Bacillus cereus: A Comparison of a Parent Strain, an Asporogenic Variant and Cell Fractions

S ummary . The antigenic structure of a stable asporogenic variant of the M8 strain of Bacillus cereus has been compared with that of the parent strain. Ultrasonic extracts of cells of both parent strain and variant harvested at different ages have been analysed by immunoelectrophoresis against antisera prepared by injecting such extracts into rabbits.
Disintegrates of cells of the asporogenic variant were antigenically identical with disintegrates of vegetative cells of the parent strain. Disintegrates of cells in later stages of sporulation and of mature spores of the parent strain contained thermostable antigens which were never detected in the variant. Antigens of isolated cell walls, protoplasts and flagella were also studied.
Examination of esterase and catalase content of the two strains showed that although the variant had the same enzymes as the young vegetative cells of the parent strain it never developed the thermostable catalase found in disintegrated spores. Protein components of the two strains at different stages of growth and of the isolated cell fractions were studied by electrophoresis in polyacrylamide gels.     

Streptomyces species producing the streptovaricin complex

Morphological, cultural and physiological-biochemical properties ofStreptomyces sp. strain 1000 and its antibiotic production were investigated. Antibiotics 1011 (identical with the streptovaricin complex) and 1012 (with antibacterial action) were isolated from the cultural broth of this strain. The overproducing natural variant 1011 was isolated from the population of a strain producing antibiotic 1011 at a concentration of 1000 mg/L (activity of the parent strain represents 41 mg/L only). Comparative taxonomical characteristic ofStreptomyces sp. strain 1000 with strains fromS. spectabilis showed that the strain 1000 differed in some properties and antibiotic production being considered as a new variant ofS. spectabilis. The strain shows an expressed antibiotic activity against G+ as well as G− bacterial and yeasts.     

Studies on a Morphologically Distinct Colchicine-Resistant Variant of Entamoeba sp.*

Colchicine has a temperature-dependent cytotoxic effect on Entamoeba sp. (Laredo isolate) that is most apparent when the drug is applied during the initiation of cultures at a concentration of 7.5 mM or higher. Continued transfer of cultures in medium containing progressively increasing concentrations of colchicine has resulted in a variant that grows prolifically in the presence of colchicine (7.5 mM) with a generation time comparable to that of the parent stock. Comparison of a number of parameters of the 2 variants revealed that colchicine resistance was accompanied by a change in cell shape, a reduced membrane permeability, which could partially be overcome by the addition of dimethyl sulfoxide (DMSO), and a reduced tolerance to osmotic stress. However, the parent strain and resistant variant were equally susceptible to cycloheximide and puromycin suggesting that the acquired colchicine resistance may not be explained on the basis of an entirely unspecific generalized reduced ability for drug uptake. Colchicine resistance and altered structure were found to be stable over a long period of time. The possible interdependence of these 2 parameters and their relation to cell motility in Entamoeba sp. are discussed.     

New types of acetate-oxidizing,sulfate-reducing Desulfobacter species,D. hydrogenophilus sp. nov., D. latus sp. nov., and D. curvatus sp. nov.

Sulfate-reducing bacteria with oval to rod-shaped cells (strains AcRS1, AcRS2) and vibrio-shaped cells (strains AcRM3, AcRM4, AcRM5) differing by size were isolated from anaerobic marine sediment with acetate as the only electron donor. A vibrio-shaped type (strain AcKo) was also isolated from freshwater sediment. Two strains (AcRS1, AcRM3) used ethanol and pyruvate in addition to acetate, and one strain (AcRS1) grew autotrophically with H₂, sulfate and CO₂. Higher fatty acids or lactate were never utilized. All isolates were able to grow in ammonia-free medium in the presence of N₂. Nitrogenase activity under such conditions was demonstrated by the acetylene reduction test. The facultatively lithoautotrophic strain (AcRS1), a strain (AcRS2) with unusually large cells (2×5 m), and a vibrio-shaped strain (AcRM3) are described as new Desulfobacter species, D. hydrogenophilus, D. latus, and D. curvatus, respectively.     

Isolation of infective and effective Frankia strains from root nodules of Alnus acuminata (Betulaceae)

Two Frankia strains were isolated from root nodules of Alnus acuminata collected in the Tucumano-oranense forest, Argentina. Monosporal cultures were obtained by plating a spore suspension of each strain and isolating a single colony. The strains (named AacI and AacIII) showed branched mycelia with polymorphic sporangia and NIR-vesicles. They differed in their ability to use carbon sources: the AacI strain grew well on pyruvate, while the AacIII strain grew on mineral medium supplemented with glucose or, alternatively, with sucrose. The two strains were sensitive to oleandomycin, erythromycin, kanamycin, penicillin G, streptomycin and chloramphenicol at 5 μg/ml. The AcIII strain exhibited a moderate resistance to rifampicin, ampicillin and vancomycin. The nitrogenase activity in vitro of the strains was significantly higher in basal medium without nitrogen than that determined in the presence of ammonium chloride. Both strains were infective on seedlings of Alnus glutinosa, inducing an approximately similar percentage of nodulated plants (80%), although strain AacIII produced a higher number of nodules per plant (≤15) than strain AacI (≤6). They were also effective for nitrogen fixation in planta, determined by the acetylene reduction assay. This revised version was published online in July 2006 with corrections to the Cover Date.     

Estudio de algunas propiedades biológicas de varias cepas de Coccidioides immitis Stiles, 1896

Summary The assimilation of sugars and nitrogenous compound by the speciesCoccidioides immitis has been studied and also some others biological propierties: proteolytic and amilolytic activity.Three strains namedCoccidioides immitis (one of then isolated in Bolivia by Dr. Veintemillas) and the strain namedTrichosporon proteolyticum were studied. The four cultures showed the same properties. Urea, asparagine and peptone are well utilized, ammonium sulphate is also utilized but potassium nitrate is not assimilated. Glucose is utilized but maltose, sucrose, galactose and lactose are not assimilated. All the strains showed proteolytic activity on the milk and gelatine but the proteolysis of the cogulated serum was not evident.     

Two new species of Haplopappus (Astereae, Asteraceae) from Mendoza, Argentina

A review of the Argentinean specimens ofHaplopappus Cass. has revealed two new species from Mendoza province, both of the sectionHaplopappus:H. boelkei, unique by its caespitose habit and long petiolate leaves, andH. mendocinus, related toH. macrocephalus (Less.) DC.,H. ochagavianus Phil. andH. tiltilensis Phil.