Suckling mouse cataract agent is a helical wall-free prokaryote (spiroplasma) pathogenic for vertebrates.
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Biomedical subjects
Publications and source records attributed to R F Whitcomb.
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The helical wall-free microorganism, Spiroplasma citri, which is associated with citrus stubborn, a disease with no known vector, multiplied in the leafhopper vector of corn stunt but multiplied to higher titer in the vector of aster yellows and decreased the longevity of that insect. Acholeplasma laidlawii and A. granularum also multiplied in both leafhoppers.
Antibiotics suppressed development of aster yellows (AY) disease symptoms in plants of china aster [Callistephus chinensis (L.) Nees.] and annual chrysanthemum (Chrysanthemum carinatum, Schousb.). When inoculated chrysanthemum plants were treated by any of several techniques with tetracycline antibiotics or chloramphenicol, symptoms failed to appear during treatment but appeared 1 to 4 weeks after treatments were terminated. Under continuous administration of chlortetracycline, aster plants with AY symptoms developed symptomless axillary growth, including flowers. Streptomycin, oleandomycin, kanamycin, tylosin, carbomycin, polymyxin, bacitracin, neomycin, sulfanilamide, penicillin, vancomycin, or cycloserine had no discernible effect on development of AY symptoms. Treatment of plants with tetracycline antibiotics before exposure to inoculative (pathogen-transmitting) vectors delayed the appearance of symptoms or prevented AY infection. Remission of AY symptoms in inoculated plants treated with chlortetracycline was correlated with an inhibition of multiplication of AY agent, as measured by bioassay of extracts. The data give additional support to the hypothesis that aster yellows disease is caused by a mycoplasma-like microorganism.
Chlortetracycline or chloramphenicol (but not kanamycin, penicillin, or erythromycin), when administered in hydroponic solution to diseased aster, reduced the availability of the aster yellows (AY) agent to nymphs of Macrosteles fascifrons (Stål). Insects exposed to healthy plants whose roots were immersed in chlortetracycline were able to acquire AY agent from diseased plants the day after removal from the antibiotic-treated plants, but the latent period of the ensuing disease in the insects was prolonged. Chlortetracycline or tylosin tartrate blocked AY infection in nymphs injected with a mixture of antibiotic and the AY agent, but polymyxin, neomycin, vancomycin, penicillin, carbomycin, or chloramphenicol did not. All tetracyclines tested, methacycline, oxytetracycline, and chlortetracycline, produced a dramatic reduction in the ability of infected vectors to transmit AY agent. Tylosin tartrate also reduced transmission when injected into AY-transmitting vectors, but carbomycin, spectinomycin, cycloserine, penicillin, erythromycin, or kanamycin had no such effect. During the first 10 days after injection of tylosin tartrate or oxytetracycline into transmitting vectors, ability of the insects to transmit AY decayed rapidly. Transmission by insects injected with buffer alone, after decreasing the first day after injection, gradually returned to its normal level in less than 1 week. By 2 to 3 weeks after injection with tylosin or oxytetracycline, ability to transmit AY was regained by vectors. The results suggest that tetracycline antibiotics and tylosin tartrate inhibit multiplication of AY agent in the insect. The spectrum of antibiotic activity in the insect is consistent with the hypothesis that AY and other plant yellows diseases are caused by mycoplasma-like organisms.
Suppression of symptoms of aster yellows by antibiotics supports the tentative hypothesis that the etiologic agent is a mycoplasma-or bedsonia-like organism rather than a virus. Development of symptoms was supressed by chlortetracycline, tetracycline, or chloramphenicol, but not by penicillin. When plants were treated with chlortetracycline at 1000 parts per million before symptoms appeared, symptoms developed only after cessation of the treatment. Assay of the agent of aster yellows, extracted from plants, indicated inhibition of growth of the pathogen by treatment with chlortetracycline. Plants severely affected before treatment began developed new symptomless axillary growth, including flowers; previously yellowed leaves often became green. Acquisition of the agent of aster yellows by leafhoppers was drastically reduced when infected plants were treated with chlortetracycline continuously for 1 week before exposure to the vectors. Our data, and preliminary evidence from purification studies, are consistent with a possible mycoplasma-or bedsonia-like etiology of the aster yellows disease.
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Acholeplasmas, spiroplasmas and other non-helical sterol-requiring mycoplasmas of unknown phylogenetic affinity inhabit insects. Of these, only spiroplasmas are known to be pathogenic. Group I-2 spiroplasmas, or Spiroplasma apis, especially in combination with other organisms, reduce honey-bee longevity. Plant pathogenic mycoplasma-like organisms are often found intracellularly in insects. Spiroplasmas are found predominantly in the gut lumen or haemolymph (or both) of their insect hosts. Pathogenicity of mycoplasmas is usually altered by extended passage in unusual hosts, in only one of two alternate hosts, or in culture media. Enhancement of experimental pathogenicity may occur with extended cultural passages, but maintenance of natural pathogenicity must be accomplished by continuous exposure to the usual host. Recent data provide new information on the ecology of pathogenicity. Spiroplasmas from unique habitats also tend to be unique. Spiroplasmas isolated from flowers appear to be adapted to insect species that frequent floral surfaces. Group IV spiroplasmas have been isolated from members of 4 holometabolous insect orders (including Lepidoptera), all of which visit flowers. Social or predatory insects, or insects with an "aggregation" phase in their life histories, also appear to be prone to spiroplasma infection. Some insect species which harbor spiroplasmas also carry infections of other mollicutes, some of which involve the haemolymph. Appearance of spiroplasmas in adult insects in nature is strongly affected by seasonality. Extensive tests of the host ranges of the new insect mollicutes will be required before their suitability for biological control can be evaluated.
The guanine-plus-cytosine (G + C) content of spiroplasmal DNA was calculated from the melting temperature determined spectrophotometrically and the buoyant density determined by equilibrium density gradient centrifugation in CsCl. Only two ranges of G + C values were found: 25-27 mol% and 29-32 mol%. The DNA of the following spiroplasmas has 25-27 mol% G + C: Spiroplasma citri (serogroup I-1); the spiroplasmas pathogenic to the honeybee (KC3, BC3, and B63; serogroup I-2); the corn stunt strain (E275; serogroup I-3); the tick strain 277F (serogroup I-4); the drosophila strain (serogroup II); and one group of flower spiroplasmas (serogroup III). The DNA of a second group of flower spiroplasmas (serogroup IV) and the SMCA strain (serogroup V) has a G + C content of 29-31 mol/. The classification of flower spiroplasmas into two groups on the basis of G + C content agrees well with the groupings based on serologic and protein analysis. Spiroplasmas isolated from honeybees in Morocco (B13) or froghoppers in Corsica (L89) have 29-31 mol% G + C, a value that corroborates the relatedness of these strains and the flower spiroplasmas of serogroup IV found by serologic analysis. Reannealing experiments between the vivo-labeled DNA of S. citri and unlabeled DNA of other spiroplasmas gave the following percentages of hybridization: 64% with honeybee spiroplasma DNA, 49% with corn stunt spiroplasma DNA, and 19% with tick spiroplasma 277F DNA; no significant hybridization was observed with DNA of any other spiroplasma. The taxonomic position of the tick spiroplasma 277F within serogroup I was confirmed by hybridization experiments involving [3H]DNA of this strain. The value of polyacrylamide gel analysis of DNA fragments produced by the action of EcoRI restriction enzyme on DNAs from various spiroplasmas is also discussed.
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We propose that Group I spiroplasmas be subdivided into seven, rather than four, subgroups. The seven subgroups showed remarkable homogeneity when several representative strains were compared. Hybridization reactions between DNAs of representative strains within subgroups were generally at least 90 percent, and usually at least 80 percent co-migrating cell proteins were found. In addition, when plasmid DNA was excluded, profiles of restricted DNA among strains within subgroups were very similar. In contrast, comparisons between Group I subgroups showed substantial heterogeneity. This heterogeneity was indicated by DNA-DNA hybridization reactions as low as 10-20 percent and only 10-15 percent co-migrating cell proteins. Spiroplasma citri (subgroup I-1), the honeybee spiroplasma (subgroup I-2), and the corn stunt spiroplasma (subgroup I-3) are all pathogenic organisms with more or less limited host ranges. Strains of these three subgroups have been repeatedly isolated from affected hosts. Since strains of subgroups I-2 and I-3 can be clearly differentiated from other Group I subgroups and all other spiroplasmas, the DNA-DNA hybridization reactions of the subgroups do not exceed 70 percent, and because they are important pathogens, we propose (subject to completion of standard requirements for species descriptions) that they be recognized as new species of the genus Spiroplasma.
Three serologically distinct groups of spiroplasmas have been recovered from ticks. Spiroplasma mirum strains (from rabbit ticks, Haemaphysalis leporispalustris) and Y32 group (VI) spiroplasmas (from Ixodes pacificus) are the only spiroplasmas to have a clear association with these arthropods. Group (VI) spiroplasmas are distinguished by an unusual nonhelical morphology and their capacity to hemadsorb guinea pig erythrocytes. S. mirum strains are unique in their ability to induce cataracts or lethal brain infections in a number of young vertebrates and in their virulence for the chick embryo. The 277F spiroplasma, while initially recovered from a pool of rabbit ticks (H. leporispalustris), is related by certain serological and genetic properties to spiroplasmas in the S. citri complex (serogroup I). These relationships suggest that the 277F spiroplasma may not be a natural inhabitant of the rabbit tick.
Data concerning serological classification of spiroplasmas are in good agreement, but slightly different numerical designations have been given to existing groups. It is proposed that a standardized system be adopted based on information developed mainly by the IRPCM working team on spiroplasmas. The type species (Spiroplasma citri) should be redefined to include only the agent of citrus stubborn disease (subgroup I-1). Six other subgroups, including three proposed by Bové et al. in this volume (I-5, I-6, and I-7), are members of the Group I complex. Because subgroups I-1, I-2, and I-3 (1) show significant reciprocal differences in DNA-DNA homology and two-dimensional electrophoretic protein profiles, (2) occupy exclusive habitats, (3) are each associated with important diseases, and (4) consist of clusters of very similar or identical strains, it is suggested that Latin binomials could be assigned to subgroups I-2 and I-3. It is proposed that those criteria could serve as general guidelines for consideration of subgroups for species status in the class Mollicutes. The I-4 subgroup is assigned an uncertain status, pending comparisons with the LB-12 (I-5), M55 (I-6), and N525 (I-7) subgroups. To previously described serogroups we add the CN-5 Cotinus beetle spiroplasma (IX), the AES-1 mosquito strain (X), and the MQ-4 Monobia strain (XI).