Chlamydia culture service.
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Chlamydia Infections↗
Biomedical subjects
D Taylor-Robinson
Publications and source records attributed to D Taylor-Robinson.
One-step staining of Neisseria gonorrhoeae in urethral discharge by methyl green-pyronin.
Methyl green-pyronin (MGP) was used in a one-step procedure to stain smears of urethral discharge from 169 men. Duplicate smears were stained by Gram's method and discharge was cultured for Neisseria gonorrhoeae. The organisms were isolated from 67 specimens and intracellular diplococci were seen in 74 smears after Gram staining and in 77 after staining by MGP. Furthermore, more extracellular and intracellular diplococci were seen in smears stained by MGP than by Gram's method and the proportion of polymorphonuclear leucocytes found to contain the organisms was greater after staining with MGP. Staining with MGP is simple, rapid, inexpensive, and easily automated.
Humans↗
A comparison of the association of Neisseria gonorrhoeae with human and guinea-pig genital mucosa maintained in organ culture.
Organ cultures of human and guinea-pig genital mucosa were inoculated with Neisseria gonorrhoeae, and the association of the bacteria with the epithelial surface of each tissue was studied by light microscopy and by scanning electron microscopy. Gonococci attached to the mucosa of human fallopian tube, adhering specifically to the surface of non-ciliated epithelial cells. In contrast, gonococci rarely attached to the mucosal surface of guinea-pig uterine horn, vagina or bladder, although organisms were occasionally seen associated with the submucosal tissue in areas where the epithelium had sloughed, and in extracellular mucus secretions. There is no evidence from this study that gonococci adhere to guinea-pig tissue in a manner analogous to that seen with human genital tissue.
Animals↗
Experimental infection of the marmoset genital tract with Chlamydia trachomatis.
A strain of Chlamydia trachomatis isolated in McCoy cells from the urethra of a patient suffering from non-gonococcal urethritis was inoculated into the vagina of 8 female marmosets. Chlamydiae were isolated repeatedly for 10-42 days from the lower genital tract of 7 of the marmosets. Six of the infected animals developed an acute inflammatory reaction in the genital tract and chlamydial inclusions in epithelial cells were seen in smears from 2 of them. In addition, each of 6 infected marmosets examined developed humoral antibodies to C. trachomatis. In contrast, 3 control animals inoculated intravaginally with chlamydia-free McCoy cells showed no evidence of chlamydial infection. Since the marmoset is small and easily bred in captivity, it should provide a useful model for studying the mechanisms of chlamydial pathogenicity.
Animals↗
Detection of antibody against Mycoplasma mycoides subsp mycoides in cattle by an enzyme-linked immunosorbent assay.
The results of an enzyme-linked immunosorbent assay (ELISA) For the detection of antibody against Mycoplasma mycoides subsp mycoides are presented. Antibody was detected in the sera of cattle at least 19 months after recovery from an infection and at least 23 months after vaccination. Almost half the sera of some animals in an area of Nigera where contagious bovine pleuropneumonia is enzootic contained antibody. Antibody was rarely detected when the same sera were examined by other established serological tests, emphasising the sensitivity of the ELISA.
Animals↗
Low frequency of chlamydial antibodies in patients with Crohn's disease and ulcerative colitis.
Serum samples from 55 patients with Crohn's disease and from 23 patients with ulcerative colitis were tested for antibodies to Chlamydia trachomatis immunotypes by a micro-immunofluorescence technique. Antibody titres of 1:8 or greater against several immunotypes were detected in 14.5% of patients with Crohn's disease and in 21.7% of those with ulcerative colitis. These figures resemble the incidence in a healthy, non-venereal-disease population. Furthermore, there was no correlation between the presence of antibody and such factors as duration of symptoms, localisation of disease, or disease activity. These findings indicate that there is no reason to believe that Crohn's disease involves chlamydiae or that examination for chlamydial antibody is helpful in diagnosis.
Adult↗
Serological response of patients with non-gonococcal urethritis to causative organism of contagious equine metritis 1977.
The presence of allugtinins to the causative organism of contagious equine metritis (C.E.M.) in human serum has been confirmed. Agglutinins were found in the serum of 84 (37.6%) of 223 patients with non-gonococcal urethritis (N.G.U.), and in 12.5% of these patients there was a four-fold or greater rise in titre during the course of their illness. There was no evidence that these agglutinins were the result of infection by chlamydiae or ureaplasmas. Certain patients with these agglutinins seemed to respond better to therapy with antibiotics to which the C.E.M. bacterium is susceptible in vitro than did patients in whom these agglutinins were not found. The findings suggest that the C.E.M. bacterium or a microorganism related to it may be aetiologically involved in a proportion of patients with N.G.U. A search for such an organism in these patients is in progress.
Agglutination Tests↗
The effect of reducing and other agents on the motility of Treponema pallidum in an acellular culture medium.
The maintenance of Treponema pallidum motility was investigated in an acellular medium based on T. pallidum immobilization test medium. The acellular medium contained cysteine, glutathione, thioglycollate and dithiothreitol as reducing agents and had a redox potential of -275 +/- 25 mV at pH 7.3. In an atomosphere containing 3% O2, motile treponemes survived four times longer when calf serum and bovine serum albumin were added to the medium. The selective omission of glutathione and, particularly, thioglycollate prolonged the survival of motile treponemes almost fivefold. In addition, stored medium, in which thioglycollate had become inactive, sustained motile treponemes for longer than did freshly prepared medium. Thus, thioglycollate is toxic for the organisms. It may be omitted from the medium because low redox potentials can be achieved without it.
Culture Media↗
Interaction of Mycoplasma pneumoniae with human lung fibroblasts: characterization of the in vitro model.
The interaction of pathogenic Mycoplasma pneumoniae and host cells was studied in cell cultures of MRC-5 human lung fibroblasts. A comparison of results obtained with fibroblasts in a monolayer format and with hamster tracheal explant cultures indicated that the former can bind significantly larger numbers of mycoplasmas. In addition, the attachment was 96% specific, that is, mediated through a neuraminidase-sensitive receptor on the host cell. Uptake of mycoplasmas was directly related to the number of mycoplasma cells present in the inoculum, and attachment was virtually complete within a 30-min period at 37 degrees C. High doses of M. pneumoniae induced a marked cytopathic effect, whereas doses of less than or equal to 10(6) colony-forming units per ml produced grossly observable cell damage that was moderate and variable. Transmission electron microscopy studies indicated that attachment of M. pneumoniae to the surface of lung fibroblasts occurred with the specialized terminal structure or binding site oriented closest to the epithelial cell surface. The filamentous mycoplasma cells were spatially arranged in several configurations and were not limited to a vertical orientation. The advantages and disadvantages of human lung fibroblast monolayer cultures, in reference to other in vitro models are discussed. A new mycoplasma agar medium (G-200 agar) with a defined tissue culture base and 10% horse serum is also described.
Binding Sites↗
Interaction of Mycoplasma pneumoniae with human lung fibroblasts: role of receptor sites.
The biochemical nature of the neuraminidase-sensitive Mycoplasma pneumoniae receptor site on human lung fibroblast cells was studied. Purified, mixed sialoglycolipid (ganglioside) preparations from human and bovine tissues did not bind to M. pneumoniae organisms and block their subsequent attachment to fibroblasts. Fibroblasts incubated for 24 h in sialoglycolipid solutions to increase the ganglioside content of their membranes did not show increased pathogen attachment when later incubated with mycoplasmas. HeLa cells grown in the presence of sodium butyrate to increase GM3 ganglioside levels likewise did not have significantly increased uptake of M. pneumoniae organisms. Treatment of fibroblasts with enzymes indicated that the mycoplasma receptor site is trypsin and papain resistant but Pronase sensitive. Pronase digests of fibroblast membranes contained a product(s) which combined with M. pneumoniae cellls and cosedimented with them during centrifugation. Glycoproteins, purified from fibroblast membranes by a lithium diiodosalicylate solubilization technique, similarly bound to M. pneumoniae organisms. Collectively, these data suggest that the major component of the M. pneumoniae receptor site is a sialoglycoprotein with little or no lipid.
Binding Sites↗
Comparison of various McCoy cell treatment procedures used for detection of Chlamydia trachomatis.
McCoy cells treated in six different ways, in addition to untreated cells, were compared to determine which gave rise to the largest number of Chlamydia trachomatis inclusions when tested with a laboratory-passaged strain. The same batch of cells was treated by irradiation, preinoculation exposure to cytochalasin B or 5-iodo-2'-deoxyuridine, and postinoculation exposure to cycloheximide, hydrocortisone, or emetine. Significantly more inclusions were always found in cells which had been treated with cycloheximide than in cells treated in any other way. Conversely, untreated McCoy cells always had significantly fewer inclusions than cells which had received some form of treatment. Similar results were obtained when cycloheximide-treated, irradiated, and untreated cells were inoculated with urethral specimens containing unpassaged organisms.
Bacteriological Techniques↗
Ureaplasma urealyticum and Mycoplasma hominis in chlamydial and non-chlamydial nongonococcal urethritis.
Urethral specimens from 726 patients with nongonococcal urethritis (NGU) were examined for Chlamydia trachomatis, Ureaplasma urealyticum, and Mycoplasma hominis. Chlamydiae were isolated from 35.9% of ureaplasma-positive patients and from 36.5% of ureaplasma-negative patients. Ureaplasmas were isolated from 52.5% of chlamydia-positive patients and from 53.1% of chlamydia-negative patients, an observation which contrasts with that of some workers who have suggested that ureaplasmas are significantly associated with chlamydia-negative NGU. Furthermore, the numbers of ureaplasmas isolated from patients who did or did not harbour chlamydiae were not significantly different nor was there a particular association of ureaplasmas with chlamydia-negative NGU in patients experiencing their first episode of disease. In addition, M. hominis was not isolated more frequently from those from whom chlamydiae were or were not isolated. The only significant associations were the isolation of M. hominis from patients who were ureaplasma-positive and of ureaplasmas from those who were M. hominis-positive. These findings do not necessarily mitigate against ureaplasmas being responsible for some cases of chlamydia-negative NGU.
Chlamydia trachomatis↗
Treatment of nongonococcal urethritis with rifampicin as a means of defining the role of Ureaplasma urealyticum.
The results of a double-blind therapeutic trial on 217 men with nongonococcal urethritis (NGU) show that minocycline was more effective than rifampicin. Before treatment Chlamydia trachomatis was isolated from 43% of men, Ureaplasma urealyticum from 59%, and Mycoplasma hominis from 22%. Chlamydiae and ureaplasmas were isolated less frequently from men with a recent history of NGU. Minocycline was given to 94 patients, and after treatment chlamydiae were isolated from only one of 40 initially chlamydia-positive patients and ureaplasmas from only five of 57 initially ureaplasma-positive patients. Although most patients responded clinically, failure and partial recovery rather than complete recovery were observed more often among those who were infected with ureaplasmas. Rifampicin was given to 123 patients, after which chlamydiae were isolated from only one of 53 initially chlamydia-positive men whereas ureaplasmas, insensitive to the antibiotic in vitro, were isolated from 55 of 68 men who had initially positive results. Patients infected with ureaplasmas failed to respond to rifampicin treatment significantly more often than those who were not infected. This was also observed when only patients who had never had NGU or who had not had a recent episode were considered. Furthermore, 24 (44%) of the 55 men whose ureaplasmas persisted failed to recover whereas only one (7·7%) of 13 men whose ureaplasmas disappeared did not respond to treatment. These results suggested that ureaplasmas were a cause of urethritis in some of the men (an estimated 10% at least). In addition, Reiter's disease developed in two men treated with rifampicin from whom only ureaplasmas had been isolated initially. M. hominis did not seem to have an important pathogenic role in NGU and there was evidence that ureaplasmas were an unlikely cause of urethritis in some men since the organisms persisted despite complete clinical recovery.
Chlamydia trachomatis↗
Growth and effect of chlamydiae in human and bovine oviduct organ cultures.
Organ cultures of 10 Fallopian tubes were inoculated with a genital strain of Chlamydia trachomatis and seven were infected. Infection was enhanced by centrifuging the organisms on to the tissues, larger numbers of organisms being reisolated from the tissues after this procedure. There was evidence of chlamydial multiplication because the number of organisms which were recovered from the tissues three to five days after inoculation had increased. Recovery was rare, however, after the sixth day, thus suggesting a self-limiting infection. Organ cultures of two bovine oviducts were infected with the bovine abortion strain of Chlamydia psittaci, but in these experiments centrifugation of the inocula did not enhance infection. The organisms were found in both the tissue and medium of cultures up to 18 days after inoculation and in much greater numbers than in the C. trachomatis-infected Fallopian cultures. Chlamydial infection was not entirely host-tissue specific, because C. trachomatis organisms were isolated from bovine oviduct cultures. Inclusions, however, were not detected histologically or electron microscopically in the epithelium of C. trachomatis-infected cultures, but they were detected by these means in C. psittaci-infected bovine cultures. All the elements of the chlamydial growth cycle were seen by electron microscopy, organisms being found in ciliated and possibly non-ciliated cells, and shedding of some infected epithelial cells was observed. No evidence of extensive epithelial cell damage was observed, however, and no loss of ciliary activity was detected in cultures infected with either C. trachomatis or C. psittaci when compared with uninoculated cultures. Thus acute salpingitis, when caused by chlamydial infection, is probably immunologically mediated.
Animals↗
Ureaplasmas in the marmoset (Callithrix jacchus): transmission and elimination.
All adult marmosets tested had ureaplasmas in their throats but not in the lower respiratory tract, and rarely in the genital tract. Ureaplasmas persisted in the throat of a marmoset separated from the colony for 44 days. They could not be recovered from the animals for at least nine weeks after a course of minocycline. Airborne reinfection did not occur when these animals were surrounded by, but separate from, infected marmosets. It occurred when the minocycline-treated animals were caged with the infected marmosets or were inoculated. The genital tract was more difficult to infect than the oropharynx.
Animals↗
Pathogenicity of ureaplasmas for animals and man.
Since their original isolation from the genital tract of man, ureaplasmas, previously termed T-strain mycoplasmas, have been isolated from a variety of animal species. Under experimental conditions they have been shown to cause mastitis in cattle, goats and mice, and observations made on naturally-occurring bovine pneumonia, as well as the results of experimental inoculation, suggest that ureaplasmas are responsible for a portion of bovine cuffing pneumonia. Ureaplasmas have been isolated from the genital tract of a wide variety of animals and have the potential for causing disease in this anatomical area, as the results of experimental intra-urethral inoculation of goats, for example, indicate. However, there are no data, as yet, to incriminate ureaplasmas as a cause of naturally-occurring genital tract disease nor as a cause of infertility, the latter being an area in which the results of studies are often conflicting and difficult to interpret. In man, the rôle of ureaplasmas in genito-urinary disease has been a bone of contention for many years. Experimentally, ureaplasmas produce bladder calculi in rats but so far there is no evidence that they do so in man. Further, there are no convincing data to support the notion that infertile couples possessing ureaplasmas should be treated with tetracyclines. There is an undoubted association between spontaneous abortion and low birth-weight on the one hand and the presence of ureaplasmas in the mother, abortus or infant on the other. However, evidence that the organisms cause abortion is lacking and whether they are directly responsible for low birth-weight is unknown. The association between chorioamnionitis and ureaplasma isolation is provocative enough to stimulate further work. In the case of non-gonococcal urethritis, the weight of evidence suggests that ureaplasmas cause the disease in some men. This is based on quantitative isolation, volunteer inoculation, as well as treatment studies including the use of antibiotics, such as rifampicin, which differentiate between chlamydiae and ureaplasmas.
Animals↗
Mycoplasmal (ureaplasma) septic arthritis in hypogammaglobulinaemia.
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Adult↗
The incidence of tetracycline-resistant strains of Ureaplasma urealyticum.
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Drug Resistance, Microbial↗