Strain Tab4cT, a helical prokaryote that was isolated from the body of a Haematopota sp. fly collected in Champchevrier, Indre-et-Loire, Touraine, France, was found to be a member of the class Mollicutes. The cells of strain Tab4cT were small, motile helices that were devoid of a cell wall. The organism passed through filters with mean pore diameters as small as 0.20 mm. Strain Tab4cT grew rapidly in liquid SP-4 medium at both 30 and 37 degrees C. The organism fermented glucose but did not hydrolyse arginine or urea, and did not require serum for growth. In preliminary electrophoretic analyses, the cell protein patterns of strain Tab4cT were distinct from those of 14 other spiroplasmas found in mosquitoes, deer flies and horse flies from Europe and the Far-East. In reciprocal metabolism inhibition and deformation serological tests, employing antigens and antisera representative of spiroplasma groups I-XXXIII (including all sub-groups), plus ungrouped strains BARC 1901 and BARC 2649, no serological relationship with Tab4cT was found. The G + C content of the DNA of strain Tab4cT was about 25 +/- 1 mol% and its genome size was 1.305 kbp. It is proposed that spiroplasma strain Tab4cT be assigned to group XVII (presently vacant) and that strain (ATCC 700271T) is the type strain of a new species, Spiroplasma turonicum.

Initially, strain CUAS-1T (T = type strain), which was isolated from a frozen triturate of Culex annulus mosquitoes collected in Taiwan, was thought to be a member of spiroplasma group VII. This placement was based on the spiroplasma deformation test titer observed when strain CUAS-1T spiroplasmas were tested with Spiroplasma monobiae MQ-1T antiserum. The results of subsequent reciprocal spiroplasma deformation, metabolism inhibition, and growth inhibition tests clearly revealed that strain CUAS-1T is not serologically related to previously described spiroplasma groups (groups I to XXIV) and thus is a representative of a new group, group XXV. Strain CUAS-1T was characterized by using the minimal standards for mollicute species descriptions. During logarithmic-phase growth, strain CUAS-1T cells are characteristically very short helices with 1.5 to 2 helical turns (1 to 2 microns), highly motile, and bounded by a single trilaminar membrane and form granular colonies with satellites when the organism is grown aerobically on MID medium containing 1.6% agar. Growth in MID broth occurs at temperatures ranging from 10 to 37 degrees C, and the optimum temperature is 30 degrees C. Substrate utilization tests revealed that cholesterol is required for growth, that glucose is hydrolyzed, and that arginine is not hydrolyzed both in the presence and in the absence of glucose. The genome of strain CUAS-1T is 1,080 kbp long, and the guanine-plus-cytosine content is 26 +/- 1 mol%. On the basis of the results of our studies we propose that strain CUAS-1T (group XXV) should be placed in a new species, Spiroplasma diminutum. Strain CUAS-1 (= ATCC 49235) is the type strain of S. diminutum.

We describe the cloning, sequencing and expression of the 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) gene of Pseudomonas testosteroni. A genomic library of P. testosteroni total DNA constructed from SauIIIA digests ligated to an lambda gt11 vector was probed with a polyclonal antibody raised against purified enzyme. Subclones derived from a recombinant phage containing a 1746 bp insert were sequenced and found to contain an open reading frame of 696 bp that corresponds to a protein of 231 amino acid residues. A search for homologous proteins was performed. No similarity was observed when comparing 3 alpha-HSD with known members of the short-chain dehydrogenase family. However a small proteic fragment (80 amino acids) shows homology with the N-terminal sequence of bacterial L7/L12 ribosomal proteins.

A genomic library of Pseudomonas testosteroi total DNA constructed from SauIIIA digests ligated to a lambda gt11 vector was probed with different polyclonal antibodies raised against purified 3 alpha-HSD and (3 beta-17 beta)-HSD. Two different clones reacting with one antibody were selected. The clone reacting with (3-17)beta-HSD antibody contained a 2,661-base pair insert and was found to contained an open reading frame of 765 base pair that corresponds to a protein of 254 amino-acid residues. A 1,492-base pair was inserte in pBR 322 plasmid vector; the recombinant bacterie over expressed the (3-17)beta-HSD gene. The clone reacting with 3 alpha-HSD antibody contained a 1746 base pair insert which contained an open reading frame of 696 base pairs that corresponds to a protein of 231 amino-acid residues. A search for homologous proteins was performed. Distant similarities were found between (3-17)beta-HSD and members of the short-chain alcool dehydrogenase (SCAD) family but no similarity was observed between 3 alpha-HSD and proteins of this family.

Some group XVI spiroplasmas, such as strains CC-1 (Spiroplasma cantharicola) and CB-1, are associated with cantharid beetles. Fifteen related but heterogeneous strains have been isolated from mosquitoes, other insects, and a flower in France and the United States. In the present study, these seventeen strains have been compared by deformation and metabolism inhibition serological tests, by one-dimensional protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and by determination of the guanine-plus-cytosine content of their DNA. Five of the 17 strains were further compared by DNA-DNA hybridization and by restriction enzyme (EcoRI and HindIII) analysis of their DNA. On the basis of the resulting data, we propose that group XVI be subdivided into three subgroups. Subgroup XVI-I is represented by strain CC-1 (ATCC 43207) from a cantharid beetle in the United States, and strain MQ-6 from a wasp; subgroup XVI-2 is represented by strain CB-1 (ATCC 43208) from a cantharid beetle and two strains from mosquitoes, all in the United States; and subgroup XVI-3 is represented by strain Ar-1357 (ATCC 51126) and contains 11 strains from mosquitoes and 1 strain from a flower, all from the Savoy region of France.

We describe the cloning, sequencing and overexpression of the (3-17)beta hydroxysteroid dehydrogenase gene of Pseudomonas testosteroni. A genomic library of Ps. testosteroni total DNA constructed from SauIIIA digests ligated to a lambdagt11 vector was probed with polyclonal antibody raised against purified enzyme. Subclones derived from a recombinant phage containing a 2661-base-pair insert were sequenced and found to contain an open reading frame of 765 base pairs that corresponds to a protein of 254 amino acid residues. A 1492-base-pair fragment was inserted into pBR322 plasmid vector and used to construct a strain of E. coli HB101 that overexpressed the steroid dehydrogenase gene.

Spiroplasma strain CC-1T, isolated from the gut of the soldier beetle Cantharis carolinus, was serologically distinct from other spiroplasma species, groups, and subgroups. Cells of strain CC-1T were shown by light microscopy to be helical, motile filaments. Electron microscopy showed that the cells were bounded by a single cytoplasmic membrane, with no evidence of a cell wall. The organism was insensitive to penicillin. Strain CC-1T grew well in SM-1, M1D, and SP-4 liquid media under aerobic or anaerobic conditions. The strain also grew in 1% serum fraction medium. Optimal growth occurred at 32-degrees-C, with a doubling time of 2.6 h, but the strain muitiplied at temperatures of 10 to 37-degrees-C. Strain CC-1T produced acid from glucose but hydrolyzed neither arginine nor urea. The guanine-plus-cytosine (G+C) content of the DNA was 26 +/- 1 mol%. Other uncloned isolates from C. carolinus exhibited similar or identical serological patterns. On the basis of the data presented here, strain CC-1T (= ATCC 43207), previously proposed as the representative strain of subgroup XVI-1, is designated the type strain of a new species, Spiroplasma cantharicola.

Les auteurs rapportent l'observation d'un malade, agriculteur, présentant une mycose inflammatoire de la main ayant entraîné une hospitalisation. Le diagnostic n'a été fait qu'après isolement du champignon responsable : Trichophyton mentagrophytes var asteroïdes. A cette occasion, les auteurs passent en revue les problèmes diagnostiques posés par ces affections.

Spiroplasma isolates recovered from female mosquitoes (Culex tritaeniorhynchus) collected in Taiwan were found to be similar in their serological properties. Strain CT-1T (T = type strain) proved to be serologically unrelated to all currently recognized spiroplasma groups and subgroups. Strain CT-1T was studied by using criteria proposed by the International Committee on Systematic Bacteriology Subcommittee on Taxonomy of Mollicutes for the description of new mollicute species. The organisms were shown to belong to the class Mollicutes by the ultrastructure of their limiting membrane, their colonial morphology, and their filtration patterns and to the family Spiroplasmataceae by their helical morphology and motility. Growth in SP-4, M1A, and M1D media occurred at 22 to 30°C. Cholesterol was required for growth. Glucose was fermented, but arginine was not hydrolyzed. The base composition (guanine-plus-cytosine content) of the deoxyribonucleic acid of strain CT-1T was found to be 25 ± 1 mol%. On the basis of these findings, we propose that spiroplasma strains with these characteristics should be recognized as a new species, Spiroplasma taiwanense. Strain CT-1T has been deposited in the American Type Culture Collection as strain ATCC 43302T.

A spiroplasma strain Ar-1343T (T = type strain), one of a large assemblage isolated from a mixed pool of Aedes stricticus and Aedes vexans mosquitoes in the French northern Alps, proved to be serologically unrelated to all currently recognized spiroplasma serogroups (I through XI). Strain Ar-1343T was examined with the criteria proposed by the International Committee on Systematic Bacteriology Subcommittee on the Taxonomy of Mollicutes for description of new mollicute species. These spiroplasmas were shown to belong to the class Mollicutes because of their membrane ultrastructure, their colony morphology, and their filtration patterns and to the family Spiroplasmataceae because of their helical morphology and motility. Growth in SP-4, M1A, or M1D media occurred at 20 to 32°C. Strain Ar-1343T was unable to grow in the absence of cholesterol in the medium. Glucose was fermented, and arginine was catabolized. The base composition (guanine plus cytosine) of the deoxyribonucleic acid of strain Ar-1343T was found to be 30 ± 1 mol%. On the basis of these findings, we propose that spiroplasma strain Ar-1343T, with the characteristics described here, should be recognized as a new species, Spiroplasma sabaudiense. Strain Ar-1343T has been deposited in the American Type Culture Collection, Rockville, Md. (ATCC 43303).

Spiroplasmas are helical mycoplasmas that play a significant role in plant diseases. They are also found in arthropods that are likely to bite humans, such as ticks and mosquitoes. These arthropods can act as vectors and therefore may be of epidemiologic significance. Furthermore, mainly on the grounds of morphologic evidence, spiroplasmas have been incriminated in the genesis of human Creutzfeld-Jacob disease. We recovered six strains of Spiroplasma sp. from 1927 female mosquitoes. In vitro susceptibility of each strain to the following antibiotics was studied: tetracycline, oxytetracycline, doxycycline, erythromycin, chloramphenicol, rifampin, kanamycin, gentamicin and pefloxacin. Minimal inhibitory concentrations (MICs) were determined by dilution in liquid SP4 medium using microtiter plates. Plates were incubated for 24 to 48 hours at 30 degrees C. The inoculum contained approximately 5 X 10(5) CFU/ml. Each of the six strains was found to be highly susceptible to tetracycline, oxytetracycline, doxycycline, erythromycin, chloramphenicol and pefloxacin (MICs less than or equal to 0.16 microgram/ml, 0.63 microgram/ml, 0.08 microgram/ml, 0.16 microgram/ml and 0.32 microgram/ml respectively). On the opposite, the strains exhibited resistance to rifampin and variable degrees of susceptibility to kanamycin (12.5 micrograms/ml less than MIC less than 50 micrograms/ml) and gentamicin (3.12 micrograms/ml less than MIC less than 50 micrograms/ml). From our results, spiroplasmas seem to have more or less the same susceptibility to antibiotics as mycoplasmas.

Six strains of Spiroplasma sp. were isolated from Aedes mosquitoes collected in France, in the northern part of the Alps. As these isolates came from anthropophilic species of mosquitoes, the possible epidemiological and pathological importance of such observations is briefly discussed. No virus was isolated from these same mosquitoes.

Spiroplasmas are original wall-free prokaryotes which exhibit motility and a helical shape although lacking any cell wall or motile apparatus. They are known by phytopathologists because some of them are responsible for severe arthropod-borne plant diseases, such as Spiroplasma citri, the etiologic agent of "citrus stubborn disease". Other spiroplasmas are pathogenic for a number of insects: drosophila, bees and beetles. More recently spiroplasmas were also isolated from ticks, mosquitoes, horse-flies and deer-flies opening a new chapter in medical pathology. These agents are of possible great significance in Tropical Medicine.

Les auteurs rapportent deux observations d'infections pleuropulmonaires à Eikenella corrodens et font une revue de la littérature. Commensal des cavités orale et digestive, cette bactérie devient pathogène le plus souvent sur des terrains immunodéprimés. Quand elle est isolée de sécrétions broncho-pulmonaires, il faut que le prélèvement n'ait pas été en contact avec la bouche, pour qu'elle soit rendue responsable de l'infection.

Inoculation accidentelle de la bacterie par un fil de fer provoquant une plaie de la cornee. L'infection est restee localisee a l'œil et a gueri sous antibiotherapie adaptee (gentamicine)