Immunization of mice with an avirulent pseudohyphal form of Cryptococcus neoformans

Pathogenicity tests with one dysgonic and six atypical strains of Microsporum canis were carried out on guineapigs. Five of the atypical strains were laboratory mutants from dysgonic strains isolated from living hosts. As sporulation and viability varied greatly between the strains, inocula consisted of suspensions of fungal fragments of known viable count. When a sufficiently active inoculum was used, lesions and fluorescent hairs were induced in the guinea-pigs by all but one of the strains tested. In each case the strain inoculated was reisolated from the lesions in pure culture. The significance of the results is discussed in the light of the unusual nature and origin of the strains.     

Preliminary report on the isolation of a dysgonic variety ofMicrosporum canis together with the normal variety from a cattery

The isolation of a dysgonic variety ofMicrosporum canis from a large number of cats and kittens in a cattery is described. The normal variety of this fungus was isolated at the same time from the same animals. Dysgonic varieties are thought to be mutants of normal strains, but this isolation of both forms together suggests that the relationship may be more complex.     

A zoonotic ringworm outbreak caused by a dysgonic strain of Microsporum canis from stray cats

BackgroundCats are frequent carriers of Microsporum canis and veterinary students are at high risk of exposure and acquisition of the organism a la infección.ObjectivesAn outbreak of zoonotic ringworm carried by a litter of stray cats is described. Four veterinary students, four dogs, and six cats living in five separate locations were affected. All had direct or indirect contact with the infected kitten litter. We tried to identify the causal dermatophyte.MethodsConventional and mycological culture methods were used.ResultsMicroscopic features of scrapings and hairs treated with 20% KOH strongly suggested a M. canis etiology, and a diagnosis of ringworm was empirically supported by successful treatment of humans and animals. Nevertheless, cultures failed to show the expected morphology.ConclusionsCulture features of our strain are compared with those described by other authors for dysgonic M. canis strains. Epidemiological features are also discussed.     

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.     

Characterization of three strains of Capnocytophaga canis isolated from patients with sepsis

Capnocytophaga canimorsus and Capnocytophaga cynodegmi, both commensal bacteria in the oral cavities of dogs and cats, are zoonotic pathogens. In particular, C. canimorsus causes sepsis and fatal septic shock. Recently, a novel Capnocytophaga species, C. canis, was isolated from the oral cavities of healthy dogs. It is reportedly oxidase‐negative and therefore considered avirulent in humans. In the present study, three strains of C. canis were isolated from Japanese patients with sepsis. All three strains, HP20001, HP33001 and HP40001, were oxidase‐positive. Nucleotide sequence identities of the 16S rRNA gene of the three strains to the C. canimorsus type strain ATCC35979, C. cynodegmi type strain ATCC49044 and C. canis type strain LMG29146 were 96.9–97.0%, 96.9–97.0% and 99.7–99.8%, respectively. Multi‐locus sequence analysis based on seven house‐keeping genes, dnaJ, fumC, glyA, gyrB, murG, trpB and tuf, revealed that the oxidase‐positive C. canis strains isolated in Japan and oxidase‐negative strains of C. canis from canine oral cavities in Switzerland were clustered in different genetic subgroups. These results indicate that the virulence of C. canis strains in humans is associated with oxidase activity.

     

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.     

Multiple drug resistance inAerobacter aerogenes

Multiple drug resistance resulting from continued growth in various drugs in turn followed by serial subculture in the simultaneous presence of all the drugs has been studied. In this way strains ofA. aerogenes resistant, in considerable measure, to 3, 4 and 5 drugs have been obtained. Although the final state was always a compromise between the various new properties it was often a good and very effective compromise and for this reason kinetic properties of the cells such as growth rate had to be intensively followed. Tests for presence or absence of growth after a fixed interval of time would not have sufficed. At the 5-drug stage a good compromise was obtained when the agents used were streptomycin, sulphanilamide, chloramphenicol, ampicillin and 5-aminoacridine. When tretamine (triethylene melamine) replaced 5-aminoacridine as the fifth drug, the rate of growth in the various drugs both singly and in admixture was not as good.Continued subculture in sulphanilamide medium led to a condition of hypersensitivity to chloramphenicol. Ampicillin-resistant strains were more inhibited by 2-phenylethanol than the ampicillin-sensitive parent organism. Strains resistant to chloramphenicol were also resistant to tretamine and vice versa. Such cross-resistance did not occur with any of the other drugs at the concentrations used.     

Comparative evaluation of E-test and CLSI methods for Itraconazole,Fluconazole and Ketoconazole susceptibilities of Microsporum canis strains

The incidence of resistance to antifungal agents for dermatophytes is increasing, but most of the methods currently available to test the antifungal susceptibility of Microsporum canis still require standardization. The aims of this study were: (i) to evaluate the antifungal susceptibility of M. canis strains recovered from animals to ketoconazole (KTZ), fluconazole (FLZ) and itraconazole (ITZ) using a modified CLSI broth microdilution (CLSI M38-A2-BMD) and the E-test® protocols and (ii) to estimate the agreement between the methods. Tentative azole epidemiological cutoff values (ECVs) were also proposed in order to interpret the results of in vitro susceptibility tests and to establish the agreement between the E-test and CLSI BMD methods. A total of forty clinical M. canis strains from animals with skin lesions were tested, and the essential (EA) and categorical agreement (CA) between the two methods were determined. KTZ displayed the lowest MIC values, while ITZ and FLZ the highest. The ECV for KTZ and ITZ were 4 μg/ml, while those of FLZ was 64 μg/ml. Based on ECVs, about 88% of M. canis strains were susceptible to all azoles being a cross-resistance with ITZ-FLZ registered for one strain. A total of five M. canis strains showed MIC?>?ECV for FLZ using CLSI, while one strain showed MIC?>?ECV for ITZ using both tests. KTZ, ITZ and FLZ showed EA ranging from 92.5 to 95%, for all azoles and CA?>?97% except for FLZ (87.5%). The good CA between the E-test and the CLSI BMD provides evidence of the reliability of the former method to test the antifungal susceptibility of M. canis for ITZ and KTZ and not for FLZ.

     

Antifungal and marker effects of Talisia esculenta lectin on Microsporum canis in vitro

Aims: The purpose of this study was to demonstrate the usefulness of lectin obtained from Talisia esculenta (TEL) seeds as a tool to recognize and study Microsporum canis. For this purpose, we investigated the antifungal and marker action of this lectin and the relationship of these effects with the presence of carbohydrates on the structure of this fungus. Methods and Results: The in vitro antifungal activity of TEL was analysed by broth microdilution assay. In addition, TEL was assessed against the arthroconidia present on hairs obtained from infected dogs and cats. The affinity of fluorescein isothiocyanate (FITC)‐labelled TEL for macroconidia and arthroconidia of M. canis was also tested. The effects of TEL on the growth of the M. canis strains began with 0·125 mg ml^?1, and 100% inhibition was obtained with a concentration of 2 mg ml^?1. The addition of carbohydrates, especially N‐acetyl‐glucosamine and d ‐mannose, inhibited these antifungal effects. TEL was able to inhibit the growth of arthroconidial chitin‐rich forms of M. canis obtained from hairs of infected animals and strains cultured in Sabouraud agar. FITC‐labelled TEL efficiently marked macroconidial and arthroconidial forms of M. canis, as shown by fluorescent microscopy. Conclusions: These results show that the inhibitory effects of TEL on M. canis growth may be related to the interaction of lectin with the carbohydrates present at the micro‐organism’s surface, mainly d ‐mannose and N‐acetyl‐glucosamine. Significance and Impact of the Study: Talisia esculenta can be used as an important tool in the biochemical study of M. canis or as a molecule to recognize this dermatophyte in infected tissue.     

The influence of progressive growth on the specific catalase activity of human diploid cell strains. II. Effect of cellular genotype: A heterozygous strain

Human diploid cell strains develop progressively higher levels of specific catalase activity as they grow. Following subculture activity falls again. A diploid cell strain heterozygous for the gene for acatalasia I (acatalasemia) was found to develop specific catalase activity at proportionately the same rate as normal cell strains. Yet the mutant gene reduced the absolute level of specific catalase activity which the culture attained at any given point in time. In this respect the heterozygous acatalasia I strain resembles the homozygous acatalasia II strain previously reported.     

Mitogenic Effect of the Mannans from Saccharomyces cerevisiae on Mouse Spleen Lymphocytes

The DNA synthetic activities of mannans isolated from two Saccharomyces cerevisiae strains were examined in vitro using spleen cells obtained from normal or nude BALB/c strain mice. A highly branched mannan isolated from the S. cerevisiae wild type strain induced a greater increase in mitogenic activity than those displayed by the mannan of the S. cerevisiae X2180–1A-5 mutant strain which possessed fewer branching moieties. Acid-hydrolyzed wild type strain mannan with two-thirds of the molecular weight of the parent intact mannan showed weak mitogenicity. Increases in the DNA synthetic activities of nude and normal spleen cells were almost the same as that of wild type strain mannan, while nylon wool column-passed spleen cells obtained from both normal and nude mice did not show mitogenicity with this mannan. The results indicated that the mitogenic activity was responsible for the highly branched structure of the wild type strain mannan, and that this mannan is a B-cell mitogen.     

The regulation of aromatic amino acid biosynthesis in amino acid liberating mutant strains of Anabaena variabilis

Mutant strains of Anabaena variabilis which are resistant to the tryptophan analogue, 6-fluorotryptophan, liberated a wide range of amino acids although none liberated tryptophan in detectable quantities. Four strains (FT-7, FT-8, FT-9, FT-10) produced predominantly alanine together with small amounts of phenylalamine and tyrosine, strain FT-2 liberated mainly phenylalanine and tyrosine and strain FT-6 liberated mainly glutamate, NH ₄ ⁺ and several unidentified ninhydrin-positive compounds. Two forms of 3-deoxy-D-arbinoheptulosonate 7-phosphate (DAHP) synthase were identified in the parent strain, a tyrosine-sensitive form and a phenylalanine-sensitive form. In strains FT-2 and FT-6 the phenylalanine-sensitive enzyme was not detected and in strain FT-7 it was apparently deregulated with respect to inhibition by phenylalanine. No deregulation of anthranilate synthase was observed but mutant strains were found to have higher specific activities of this enzyme than the parent strain.Abbreviations chla chlorophyll a - 6-FT 6-fluorotryptophan - DAHP 3-deoxy-D-arabinoheptulosonate 7-phosphate - PEP phosphoenolpyruvate     

Stability of entomopathogenic bacteria, <Emphasis Type="Italic">Xenorhabdus nematophila</Emphasis> and <Emphasis Type="Italic">Photorhabdus luminescens</Emphasis>, during in vitro culture

The entomopathogenic nematode–bacteria complexes Heterorhabditis bacteriophora/Photorhabdus luminescens and Steinernema carpocapsae/Xenorhabdus nematophila are mass produced for use as biological insecticides. Stability of the bacterial partner in culture is essential for maintaining traits important for both biological control and production. Two geographically distinct strains of each bacterial species were isolated from their nematode partners and serially subcultured on in vitro media to assess trait stability. Subculturing resulted in a shift to secondary cell production in one P. luminescens strain and both X. nematophila strains within ten in vitro culture cycles. However, when cell phenotypic variation was controlled in X. nematophila strains by regular selection for primary variants, no trait change was detected in the primary variant after prolonged subculture. When P. luminescens cell phenotypic variation was controlled by selection for primary variants, changes in the primary variant of both strains were noted including reductions in cell and inclusion body size and inclusion body prevalence. Bacterial ability to cause lethal infections following injection into the hemocoel of Tenebrio molitor larvae declined by more than half in primary variants of one P. luminescens strain. Conversely, yield was enhanced, with the subcultured P. luminescens strains showing 53.5 and 75.8% increases in primary cell density. Field adapted traits of primary variant P. luminescens strains tend to deteriorate during in vitro culture as tradeoffs for gains in yield. In vitro producers of the P. luminescens/H. bacteriophora complex must weigh the need for superior bacterial yield against the need to preserve traits important for biological control.     

Components of fitness in a compound chromosome strain of Drosophila melanogaster

Summary The relative net fitness of a compound chromosome strain of Drosophila melanogaster was about 0.05, compared with the chromosomally normal strain from which it was derived. Based on meiotic considerations alone, the expected relative fitness was about 0.25. There were no significant differences in fertility between the compound and normal strains; the compound strain produced about 28% as many offspring as the normal strain and developed faster than the normal strain in two replicates, and slower in one replicate. The low relative fitness of the compound strain was apparently due to assortative mating, in which normal females discriminated strongly against compound males. Implications for pest control projects are dicussed.     

Selection of Ergot Alkaloid Producers by Induced Mutagenesis

Using the induced mutagenesis technique, a series of genetically modified Clavicepssp. VKM F-2609 strains that display high levels of agroclavine and elymoclavine synthesis were selected by induced mutagenesis. Compared to the parent strain, c106 displayed a 40-fold higher level of agroclavine synthesis, and c66 displayed an eightfold higher level of elymoclavine synthesis. The levels of synthesis of other alkaloids were decreased in these strains. The effects of various carbohydrates on the strain growth and ergot alkaloid biosynthesis was then investigated in both the parent strain and c106. The largest amount of agroclavine was synthesized by c106 strain growing on a medium with maltose.     

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.     

Amino acid accumulation by an analogue sensitive mutant ofCorynebacterium glutamicum

Summary A fluoroacetate/fluoropyruvate-sensitive mutant was derived from the parent strainCorynebacterium glutamicum ATCC 21513. Accumulation of various amino acids in the fermentation broth using the two strains was compared. The FA/FP-sensitive mutant accumulated about 26.5 g/L L-lysine and 2.2 g/L aspartic acid which was about 3-fold and 10-fold respectively, more than the amount produced by the parent strain.     

The HLE hybrid dysgenesis syndrome in the midgeChironomus thummi: Differences in the dysgenic potential between strains

After crossing experiments between twoChironomus subspecies the HLE hybrid dysgenesis syndrome occurs non-reciprocally in the F1 hybrids if the males of each of the fourCh. th. thummi strains (HL, So, G, Kr) are crossed to females ofCh. th. piger, strain E. In hybrid egg masses, differences in the strength of the abnormal trait reduced egg hatch were found between the differentpiger E ×thummi interstrain crosses. From the results it is concluded that eachthummi strain has a specific hybrid dysgenesis potential which determines the severity of the HLE syndrome in the hybrids. The HLE syndrome occurs also in the hybrids of crosses of differentCh. th. thummi strains. In these hybrids the syndrome is only obtained if the male parent descends from a strain which has a higher HLE hybrid dysgenesis potential than that strain from which the female parent is derived. These results demonstrate that the HLE syndrome is not restricted to the hybrids of thethummi × piger subspecies cross. In theCh. th. thummi interstrain crosses the hybrids of that cross direction are normal, which is reciprocal to the cross direction affected by the HLE syndrome. This is in contrast to thethummi × piger subspecies crosses, where the hybrids of one cross direction always show the HLE syndrome and the hybrids of the reciprocal cross direction the Rud syndrome. Thus, the present study also demonstrates that the HLE and Rud hybrid dysgenesis syndrome inChironomus are independent from one another.     

Complete Genome Sequence of Brucella canis Strain 118, a Strain Isolated from Canine

Brucella canis infects several species of animals, and canine is the preferred host. Genome sequences of strains from different hosts are valuable for comparative analysis of host adaptation and microevolution. Here, we report the genome sequence of Brucella canis strain 118, a strain isolated from canine.     

The influence of progressive growth on the specific catalase activity of human diploid cell strains. I. Effect of cellular genotype: Homozygous strains

The specific catalase activity of human diploid cell strains increases with progressive growth of the culture, and falls again following subculture. Although the increase is small, it is readily demonstrable, and is exponential with time. The response of catalase activity to proggressive growth of the culture was studied in three abnormal human cell lines. A diploid cell strain, developed from a patient homozygous for the gene causing acatalasia I, had no detectable catalase activity throughout the life cycle of the culture. Another diploid cell strain, developed from a patient homozygous for the gene causing acatalasia II, had about 5% normal catalase activity, but the proportionate increase in specific activity as the culture grew was the same as for normal cells. Thus the mutation causing acatalasia II does not change the responsiveness of the cell in terms of catalase activity to progressive growth of the culture. The behavior of a heteroploid line was similar to that of the normal diploid strains, but when the growth of the heteroploid cultures reached a plateau, their population densities were four times higher than those of the diploid strains and they had about twice the specific catalase activity.