Spiroplasma melliferum, a New Species from the Honeybee (Apis mellifera)

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Fourteen spiroplasma strains, primarily of insect origin, were analyzed according to criteria previously proposed for description of new serogroups of the genus Spiroplasma. When tested by reciprocal metabolism inhibition, growth inhibition, and deformation serological procedures, 12 of the strains were serologically unrelated to each other and to representative strains previously assigned to groups I to XI and subgroups 1-1 to 1-8. Examination by dark-field and electron microscopy indicated that each of the 12 strains possessed morphological features typical of spiroplasmas (helicity , motility, lack of a cell wall, and absence of periplasmic fibrils). All strains were resistant to 500 U of penicillin per ml and catabolized glucose but were unable to hydrolyze urea. Ability to hydrolyze arginine varied among strains. The guanine-plus-cytosine contents of the deoxyribonucleic acid of the 12 strains varied from 24 to 29 mol%. Two other strains (MQ-6 and Ar-1357) shared only a partial serological relationship to strain CC-1 (group XVI), suggesting that this group may consist of an assemblage of heterogeneous serovars. On the basis of the unique serological distinctions and other properties reported herein, we propose that the 12 representative strains be assigned new consecutive group designations XI1 to XXIII.

“…Tests for the ability of spiroplasmas to catabolize glucose and to hydrolyze arginine or urea were performed as described earlier (9,28).…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Revised Group Classification of the Genus Spiroplasma (Class Mollicutes), with Proposed New Groups XII to XXIII

Tully

Rose

Clark

et al. 1987

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“…Residence in the gut of a predaceous insect could simply reflect acquisition from prey. Some mollicutes that reside in insect hemolymph, such as SpiropZasma melliferum (7) and Spiroplasma apis (15), reduce the longevity of the host. Strain MQ-4T, like other insect gut spiroplasmas, is not known to be pathogenic to its insect host.…”

Section: Resultsmentioning

confidence: 99%

Spiroplasma velocicrescens sp. nov., from the Vespid Wasp Monobia quadridens

Konai

Whitcomb²,

Tully³

et al. 1995

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Spiroplasma strain MQ-4T (T = type strain), which was isolated from the hemolymph of the vespid waspMonobia quadridens, was serologically distinct from other spiroplasma species, groups, putative groups, and subgroups. Each strain MQ-4T cell was helical and motile and was surrounded by a single cytoplasmic membrane; there was no evidence of a cell wall. The strain grew well in 1% serum fraction medium, as well as in SM-1, MlD, and SP-4 liquid media, under both aerobic and anaerobic conditions. Strain MQ-4T grew at temperatures ranging from 10 to 41°C but did not grow at 43OC. The strain grew optimally at 37°C with a doubling time of 0.6 h, the shortest doubling time recorded for any spiroplasma. Strain MQ-4T catabolized glucose and arginine but did not hydrolyze urea. The guanine-plus-cytosine content of the DNA was about 27.5 f 1 mol%. The genome size was 1,480 kbp (940 MDa). Strain MQ-4 (= ATCC 35262) is designated the type strain of a new species, Spirophma velocicrescens.The genus Spiroplasma (31), which originally was considered a taxon that contains insect-borne pathogens of plant phloem, was later shown by Clark and his associates (6, 7) to be associated primarily with insects. As the true affiliations of members of this genus were discovered, unique features that characterized each new group emerged. However, for some time, it appeared that the group XI spiroplasmas, represented by strain MQ-4T (T = type strain), which was isolated from the gut of the predaceous wasp Monobia quadridens, might have no unusual properties. This host had yielded several other spiroplasma taxa (6), including the group VII organism Spiroplasma monobiae MQ-1 (28), and it was felt that the spiroplasmas could have been acquired from prey insects (12). However, Konai et al. (14) have recently shown that strain MQ-4T is the fastest growing spiroplasma that has been assigned to a group.In this paper we present the results of a taxonomic study of strain MQ-4T, in accordance with the proposed (13) minimal requirements for assignment of binomial names to mollicutes. We conclude that strain MQ-4, the type strain of group XI, represents a new species, Spiroplasma velocicrescens. MATERIALS AND METHODSSpiroplasma strains. Clark originally isolated strain MQ-4T by macerating the gut contents of vespid wasps (M. quadridens) collected in Maryland (6). Some of the genomic and serologic characteristics of this organism have been reported previously (3,23,27,31). Strain MQ-4T was purified by classical filtration-cloning techniques (21). Representative strains of the 23 recognized groups and eight subgroups (11,23), including the type strains of the 15 previously recognized species and seven new strains, each representing a putative new group (26, 29), were used in various parts of this study.Culture media and cultivation techniques. Strain MQ-4T was selected from a population of helical cells growing in a primary culture in SM-1 liquid medium (25) at 30°C. After several early broth passages, the isolate was lyophilized. Later, the dried culture...

“…Strain MQ-lT and isolate MQ-7 were isolated from hemolymph samples from male vespid wasps (M. quadridens). Two isolates (MQ-8 and MQ-9) serologically identical to strain MQ-lT were isolated from the guts of wasps; most mollicutes cultivated from insects are isolated from guts (7,8,15,36). Some mollicutes that reside in insect hernolymph, such as S. mellifemm (8) and S. apis (19), reduce the longevity of the host.…”

Section: Resultsmentioning

confidence: 99%

“…Two isolates (MQ-8 and MQ-9) serologically identical to strain MQ-lT were isolated from the guts of wasps; most mollicutes cultivated from insects are isolated from guts (7,8,15,36). Some mollicutes that reside in insect hernolymph, such as S. mellifemm (8) and S. apis (19), reduce the longevity of the host. Whereas S. melrifemm titers in bee hemolymph reach lo1' organisms per ml, strain MQ-lT had a titer of 7 x lo7 organisms per ml in the wasps (15).…”

Section: Resultsmentioning

confidence: 99%

Spiroplasma monobiae sp. nov. from the Vespid Wasp Monobia quadridens (Hymenoptera: Vespidae)

Whitcomb

Tully

Rose

et al. 1993

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Spiroplasma strain MQ-lT (T = type strain) from the hemolymph of the vespid wasp Monobia quadridens differed serologically from other spiroplasma species, groups, and subgroups. Cells of strain MQ-lT were helical and motile and possessed a single cytoplasmic membrane, with no evidence of a cell wall. The organism grew in conventional mycoplasma medium, in serum fraction, SM-1, MlD, and SP-4 liquid media, and on SP-4 solid medium in either aerobic or anaerobic environments. The optimum temperature for growth was 32"C, but multiplication occurred over a wide temperature range (10 to 37°C). The doubling time at 32°C in M1D medium was 1.9 h. Strain MQ-lT catabolized glucose but hydrolyzed neither arginine nor urea. Previous work showed that strain MQ-lT has a unique methylase, previously known only in eucaryotes. Also, strain MQ-lT induces production of tumor necrosis factor in bone marrow macrophages. The guanine-plus-cytosine content of the DNA was 28 f 1 mol%. The genome size of strain MQ-lT was 940 kb (627 MDa); a similar strain, MQ-8, had a genome size of 985 kb (657 MDa). Strain MQ-lT and its allies have the smallest genomes known in the genus Spiroplasma. Strain MQ-1 (= ATCC 33825) is designated the type strain of a new species, Spirophsma monobiae.The genus Spiroplasma (36), originally thought to consist of plant pathogens (22), was later shown, largely through the efforts of T. B. Clark and his associates (7,15), to be associated with arthropods. As the immense diversity of this genus unfolded, numerous interesting properties emerged. For example, group I organisms (2) were the only spiroplasmas found to be associated with plant disease or with plant-sucking bugs. Yet, the subgroup 1-2 spiroplasma, Spiroplasma melliferum (8), which was closely related to the plant spiroplasmas, was primarily associated with bees. One subgroup of this cluster (subgroup I-6), which appears to occur in the insect-plant surface habitat (15), had a guanineplus-cytosine (G+C) content that was higher than the G+C contents of its cohort subgroups (5). Another spiroplasma (subgroup I-4), isolated only once, appeared to be associated with ticks. Group I1 spiroplasmas (34) produced sex ratio abnormalities in Drosophila flies. Group 111 spiroplasmas (9) (Spiroplasma jloricola) proved to be associated with magnoliaceous flowers and their beetle and fly visitors and to have idiosyncratic metabolic pathways (6) that were readily studied because of the rapid growth of the species. Group IV appeared to be a widely distributed cluster of spiroplasmas, some of whose members, including type strain B31 (Spiroplasma apis), were pathogenic (19, 20) for honeybees @pis mellifera). Groups V (Spiroplasma mirum [30]) and VI (29) were associated with ticks. Thus, each of the first six established spiroplasma groups appeared to possess unusual biological and/or ecological properties.For some time, the group VII spiroplasmas, strain MQ-lT (T = type strain) and its allies, appeared to be exceptions to the emerging uniqueness of newly discovered strains. H...