Urethral infection with Haemophilus ducreyi in men

Carriage of Haemophilus ducreyi has not been adequately investigated in men, and its role in urethritis is unknown. In this study 456 men with H. ducreyi-culture positive genital ulcers were investigated. Among these men, 16 (3.5%) had urethritis. H. ducreyi was isolated from the urethra of nine men (1.9%). Of 57 men who had chancroid but not urethritis, none had H. ducreyi isolated from the urethra. Of 106 patients presenting with only acute urethritis, H. ducreyi was found in one. We conclude that H. ducreyi is responsible for acute urethritis in men, but that it rarely, if ever, can be cultured from the urethra of men with chancroid who do not have symptomatic urethritis.     

Chancroidal ulcers that are not chancroid. Cause and epidemiology

Hemophilus ducreyi was isolated from the ulcer of one of 33 patients in whom chancroid had been diagnosed clinically. Herpes simplex virus was isolated from 16 of these patients. A presumptive diagnosis of chancroid was made in one of three sexual partners of the only index patient with culture-proved chancroid. No chancroidal lesions were found in any of 39 sexual partners of the other 32 index patients. Genital cultures for herpes simplex virus were positive from four of 14 sexual partners of index patients with genital herpes simplex infection. The differential diagnosis of chancroidal genital ulcers is facilitated by herpes simplex virus cultures and a new selective H ducreyi culture medium.     

Comparison of media for the primary isolation of Haemophilus ducreyi

The rates of isolation of Haemophilus ducreyi from patients with presumed chancroid has been low, and improved cultural techniques are required. We determined the isolation rates of H. ducreyi from 38 patients with clinical chancroid and compared gonococcal agar supplemented with bovine hemoglobin to Mueller-Hinton agar supplemented with chocolated horse blood, each with and without 5% fetal calf serum. The rate of isolation and the qualitative and quantitative growth of strains was best on gonococcal agar with added fetal calf serum (GC-HgS). However, three strains were isolated only on Mueller-Hinton base (MH-base). GC-HgS was compared with MH-HB for the isolation of H. ducreyi from 201 patients with clinical chancroid. Seventy-one percent of cultures were positive on GC-HgS, and 61% were positive on MH-HB (P less than .005); however, 10% of cultures were positive only on MH-HB. The use of the two media together increased the yield of positive cultures to 81%. Thus maximal rates of isolation of H. ducreyi from clinical specimens requires the use of two media, GC-HgS and MH-HB.     

Treatment of chancroid. A comparison of sulphamethoxazole and trimethoprim-sulphamethoxazole.

Since sulphonamides are no longer predictably effective in the treatment of chancroid the combination of trimethoprim-sulphamethoxazole (TMP-SMX) was evaluated to identify other effective regimens. One hundred and nine patients with genital ulcers (75 men and 34 women) seen at the Special Treatment Clinic in Nairobi, Kenya, were randomly assigned to treatment with a seven day course of either sulphamethoxazole 1000 mg twice daily or trimethoprim (160 mg)-sulphamethoxazole (800 mg) (TMP-SMX) twice daily. Haemophilus ducreyi was isolated from the ulcer in 57 patients (33 men and 24 women). 16 patients were subsequently diagnosed serologically as having syphilis. No aetiological diagnosis was made in 40 patients. Treatment with sulphamethoxazole failed in five of 21 (24%) culture positive patients who were available for evaluation after seven days, whereas all 19 of such patients who were treated with TMP-SMX responded to treatment. Of the 21 isolates available for susceptibility testing, all were susceptible to trimethoprim alone (MIC less than 0.5 mg/l) and three were resistant to sulphonamides, all three containing a 4.9 megadalton (Mdal) plasmid. Two of the three patients from whom these isolates had been obtained were treated with sulphamethoxazole and both were clinical and bacteriological failures. Five of six patients with sulphonamide-susceptible H ducreyi responded to treatment with sulphamethoxazole. Failure of sulphonamides to eradicate H ducreyi in some patients with chancroid is associated with the presence of a sulphonamide resistant plasmid. In regions where this plasmid is present in H ducreyi TMP-SMX is the preferred treatment for chancroid.     

Asymptomatic carriage of Haemophilus ducreyi confirmed by the polymerase chain reaction.

OBJECTIVE--To investigate asymptomatic carriage of Haemophilus ducreyi by using polymerase chain reaction (PCR) on samples from women at high risk of infection. SUBJECTS--213 commercial sex workers (CSWs) recruited in The Gambia, West Africa. METHODS--Genital samples (cervical, vaginal and ulcer) were tested for the presence of H ducreyi by PCR with the technique of "one tube nested primer". RESULTS--12 CSWs were PCR positive for H ducreyi; 8 of these women had genital ulcers on examination. CONCLUSION--Using a simplified PCR technique for detecting H ducreyi we have shown that 2% of CSWs were carrying the organism without clinical symptoms or signs. This has important implications for sexually transmitted disease control programmes in areas with a high prevalence of chancroid.     

Rapid detection of Haemophilus ducreyi in clinical and experimental infections using monoclonal antibody: a preliminary evaluation.

A monoclonal antibody raised against Haemophilus ducreyi was tested for its sensitivity and specificity as an immunofluorescence (IF) reagent using simulated vaginal smears containing H. ducreyi, smears taken from skin lesions of mice infected with H. ducreyi and patients from South Africa, Thailand and Malaysia with clinically diagnosed chancroid. The IF test was more sensitive than culture or Gram staining in the simulated smears, theoretically detecting less than 4 organisms/sample. It detected H. ducreyi in 95% of the animal lesions compared with 14% detected by culture. Immunofluorescence testing identified over 90% of culture-positive cases of chancroid but also detected organisms in some culture-negative cases where clinical evidence for the diagnosis was strong. These results suggest that this antibody may provide a simple, rapid and sensitive means of detecting H. ducreyi in cases of chancroid.     

Chancroid: clinical manifestations,diagnosis, and management

Chancroid is a sexually transmitted disease (STD) caused by the Gram negative bacterium Haemophilus ducreyi and is characterised by necrotising genital ulceration which may be accompanied by inguinal lymphadenitis or bubo formation. H ducreyi is a fastidious organism which is difficult to culture from genital ulcer material. DNA amplification techniques have shown improved diagnostic sensitivity but are only performed in a few laboratories. The management of chancroid in the tropics tends to be undertaken in the context of syndromic management of genital ulcer disease and treatment is usually with erythromycin. A number of single dose regimens are also available to treat H ducreyi infection. Genital ulceration as a syndrome has been associated with increased transmission of human immunodeficiency virus (HIV) infection in several cross sectional and longitudinal studies. Effective and early treatment of genital ulceration is therefore an important part of any strategy to control the spread of HIV infection in tropical countries.     

Comparison of Sheffield media with standard media for the isolation of Haemophilus ducreyi

Isolation of Haemophilus ducreyi is the only method for the definitive diagnosis of chancroid. Culture on supplemented gonococcal base (GCHgs) or on supplemented Muller-Hinton agar (MHHb) has yielded the best isolation rates. Sheffield media is an alternative to standard media. We compared the isolation rate of H. ducreyi on GCHgs and MHHb to that on the Sheffield media with and without 5% horse blood. Vancomycin (3 mg/L) was added to all media. Of the 87 specimens cultured from patients with genital ulcer disease, 57 (66%) were positive on either GCHgs or MHHb or on both. Twenty six (30%) were positive on GCHgs only, 15 (17%) on MHHb only, and 15 (17%) on both. One culture was positive on Sheffield medium with 5% horse blood; none were positive on Sheffield medium without horse blood. Stock strains showed good growth at 48 hr on GCHgs and MHHb, but no growth at 48 hr and only minimally detectable growth at 72 hr on Sheffield media with horse blood. No growth was detected on Sheffield media without horseblood. Thus optimal culture of H. ducreyi in Kenya requires two media, supplemented gonococcal base and Muller-Hinton agar. Sheffield media is not useful for the primary isolation of H. ducreyi.     

Clinical and microbiologic studies of genital ulcers in Kenyan women

The etiology of genital ulcers in women in tropical regions is poorly understood. Eighty-nine women, presenting to a sexually transmitted disease clinic in Nairobi (Kenya) with a primary complaint of genital ulcers, were evaluated prospectively in a clinical and laboratory study. A final etiologic diagnosis was possible for 60 (67%) of the women. Culture for Haemophilus ducreyi was positive for 43 women, eight had secondary syphilis with ulcerated condyloma latum, three had primary syphilis, one had both chancroid and syphilis, two had moniliasis, two had herpetic ulceration, and one had a traumatic ulcer. The clinical characteristics that best distinguished chancroid from secondary syphilis were ulcer excavation and a rough ulcer base. No etiologic diagnosis was established for 29 patients. However, the clinical and epidemiologic features of these patients suggested that they were similar if not identical to the patients with H. ducreyi culture-positive chancroid. Further studies are necessary to determine the etiology of ulcers in females in whom no pathogen was identified.     

Pathogenic microbial flora of genital ulcers in Sheffield with particular reference to herpes simplex virus and Haemophilus ducreyi.

The pathogenic microbial flora of genital ulcers in 161 (80 men and 81 women) unselected patients was studied prospectively. In only one case was Treponema pallidum responsible whereas herpes simplex virus was considered to be the cause of 130 (80.8%) genital ulcers. H ducreyi was isolated from 46 (28.6%) patients, most commonly as a secondary pathogen in herpetic lesions. Two or more pathogens were isolated from the ulcers in 67 (41.6%) patients, and in 21 (13%) patients no pathogens were isolated. Our results indicate an urgent need for antiviral treatment to reduce the local reservoir of genital herpes, challenge traditional concepts about the prevalence of H ducreyi in Britain, and call for a reappraisal of its role in the causation of genital ulcers.     

Etiology of nonvesicular genital ulcers in Winnipeg

Twenty-eight patients with recent onset of nonvesicular ulcerative genital lesions were studied prospectively to define the microbial etiology. Causative agents identified included herpes simplex virus (HSV), Chlamydia trachomatis, Haemophilus ducreyi, and Treponema pallidum. Three women and 25 men (seven of whom were homosexual) were studied. Ten patients did not have an etiology identified. Nine patients had syphilis, six had genital herpes, one had chancroid, one had granuloma inguinale, and one had a human bite injury. Six of seven homosexual men had syphilis, as compared with one of eight heterosexual men (P less than .002). Among heterosexual men, five had atypical HSV infections, three with extensive balanoposthitis and two with chancroidal ulcers. Of the clinical characteristics examined, induration of the ulcer base and tenderness on palpation were predictive of etiology, whereas numbers of ulcers and regional lymphadenopathy were not. H. ducreyi was not a common cause of genital ulcers in Winnipeg, and it was not found as a commensal or superinfecting organism in ulcers caused by other agents.     

Sheffield medium for cultivation of Haemophilus ducreyi.

Our interest in the role of Haemophilus ducreyi in genital ulceration led us to examine the various media commonly used for the cultivation of the organism. We describe an improved medium for the routine isolation of H ducreyi. In comparative studies using 50 test strains originally isolated in the United Kingdom, Canada, the United States of America, and Kenya, the new medium proved superior to three standard media in requiring a shorter incubation period to first visible growth, giving larger colonies in the same period, and making possible a starch aggregation test which we have found helpful in the presumptive identification of H ducreyi from clinical material.     

Use of an adsorption enzyme immunoassay to evaluate the Haemophilus ducreyi specific and cross-reactive humoral immune response of humans.

Serodiagnosis of chancroid is limited by the cross-reactivity of Haemophilus ducreyi with Haemophilus influenzae and Haemophilus parainfluenzae. This research describes an adsorption enzyme immunoassay (EIA) that assesses the humoral immune response of North Americans and Africans to H. ducreyi. Adsorption effectively removed anti-H. influenzae and anti-H. parainfluenzae antibodies, revealing that North American control sera had no residual anti-H. ducreyi reactivity. However, African control sera still had a residual anti-H. ducreyi response. Assessment of the duration of the humoral immune response in sera from African patients with chancroid showed that the humoral antibodies persisted for up to 8 months after the diagnosis. This may explain the lack of specificity of the adsorption EIA in areas where chancroid is endemic. The detection of the humoral immune response was affected by the strain of H. ducreyi used, with indigent strains being most useful. Using H. ducreyi 35000 for Canadian sera, the sensitivity of the adsorption EIA was 100% and the specificity was 88%. For African sera, H. ducreyi strain R018 was used, and the adsorption EIA had a sensitivity of 81% and a specificity of only 23%. These data reveal that the existing humoral response in a country where chancroid is endemic differs from that in a country where it is not, and that care must be used interpreting unadsorbed humoral immune responses. The adsorption EIA approach may prove useful as an epidemiologic tool for definition of existing (past and present) levels of exposure to H. ducreyi.     

Pathogenic microbial flora of genital ulcers in Sheffield with particular reference to herpes simplex virus and Haemophilus ducreyi

The pathogenic microbial flora of genital ulcers in 161 (80 men and 81 women) unselected patients was studied prospectively. In only one case was Treponema pallidum responsible whereas herpes simplex virus was considered to be the cause of 130 (80.8%) genital ulcers. H ducreyi was isolated from 46 (28.6%) patients, most commonly as a secondary pathogen in herpetic lesions. Two or more pathogens were isolated from the ulcers in 67 (41.6%) patients, and in 21 (13%) patients no pathogens were isolated. Our results indicate an urgent need for antiviral treatment to reduce the local reservoir of genital herpes, challenge traditional concepts about the prevalence of H ducreyi in Britain, and call for a reappraisal of its role in the causation of genital ulcers.     

Genital ulcers in Kenya. Clinical and laboratory study.

Of 97 patients with genital ulcers attending a special treatment clinic in Nairobi, Kenya, 60 harboured Haemophilus ducreyi, four herpes simplex virus, and five Neisseria gonorrhoeae. Eleven patients had serological evidence of syphilis; of these one case was confirmed by darkfield microscopy. In the remaining cases no aetiological agent was identified. An enriched chocolate agar with vancomycin and serum was a useful medium for primary isolation of H ducreyi. Tetracycline was generally ineffective in the treatment of ulcers, but sulfadimidine was successful in almost 80% of cases.     

Chancroid or chancroidal ulcers

1532 cases with genital ulcers were investigated, of whom 610 presented with features suggestive of chancroid; classic, multiple lesions of chancroid were observed in 312, while its other variants, i.e. dwarf, giant and phagedaenic chancroid were also seen. In addition, 162 cases had a conspicuous morphology characterised by a single ulcer which was well-defined, soft, tender, non-indurated and had weakening edges. These were termed chancroidal ulcers. The latter had a longer incubation period of 8-11 days. Absence of lymphadenopathy was prominent in these cases. The male/female ratio was 27/1. Persons of low socio-economic status in the sexually vulnerable age-group were predominantly affected. The prepuce, coronal sulcus and glans penis were the common sites of affliction in males, while the labia minora was frequently involved in females. Due to the limited value of gram-stained smears for the detection of H. ducreyi and lack of good culture media, chancroid and chancroidal ulcers should be differentiated clinically.     

Genital ulcers in Kenya. Clinical and laboratory study

Of 97 patients with genital ulcers attending a special treatment clinic in Nairobi, Kenya, 60 harboured Haemophilus ducreyi, four herpes simplex virus, and five Neisseria gonorrhoeae. Eleven patients had serological evidence of syphilis; of these one case was confirmed by darkfield microscopy. In the remaining cases no aetiological agent was identified. An enriched chocolate agar with vancomycin and serum was a useful medium for primary isolation of H ducreyi. Tetracycline was generally ineffective in the treatment of ulcers, but sulfadimidine was successful in almost 80% of cases.     

Treatment of chancroid with erythromycin. A clinical and microbiological appraisal.

One hundred and thirty seven patients presenting with genital ulcerations from which Haemophilus ducreyi was isolated were treated with erythromycin stearate 500 mg every six hours for seven days. Of these, 91 (66%) had associated inguinal lymphadenopathy. Only two of the 100 patients who returned after one week showed no clinical improvement. Despite decrease in size H ducreyi was reisolated from the ulcers of three patients, two of whom had not complied with treatment. The patients were treated for a further week either with erythromycin or with a placebo preparation and on day 14 no discernible difference in clinical response was evident. H ducreyi was not reisolated from any lesion. In contrast, the natural course of development of associated lymphadenopathy was not modified by treatment. H ducreyi was not, however, isolated from any gland after the start of treatment. Side effects attributable to erythromycin were minimal and treatment had to be discontinued in only two patients. This study clearly indicates that treatment with erythromycin for one week results in rapid healing of lesions and the elimination of H ducreyi from both ulcers and associated lymph glands.     

Treatment of chancroid with erythromycin. A clinical and microbiological appraisal

One hundred and thirty seven patients presenting with genital ulcerations from which Haemophilus ducreyi was isolated were treated with erythromycin stearate 500 mg every six hours for seven days. Of these, 91 (66%) had associated inguinal lymphadenopathy. Only two of the 100 patients who returned after one week showed no clinical improvement. Despite decrease in size H ducreyi was reisolated from the ulcers of three patients, two of whom had not complied with treatment. The patients were treated for a further week either with erythromycin or with a placebo preparation and on day 14 no discernible difference in clinical response was evident. H ducreyi was not reisolated from any lesion. In contrast, the natural course of development of associated lymphadenopathy was not modified by treatment. H ducreyi was not, however, isolated from any gland after the start of treatment. Side effects attributable to erythromycin were minimal and treatment had to be discontinued in only two patients. This study clearly indicates that treatment with erythromycin for one week results in rapid healing of lesions and the elimination of H ducreyi from both ulcers and associated lymph glands.     

HIV and other sexually transmitted diseases at a rural hospital in Zimbabwe.

OBJECTIVE--To define the epidemiological characteristics of STD patients attending an outpatient clinic in rural Zimbabwe, to examine the aetiologic agents causing infection and to determine their relationship with HIV infection. SUBJECTS--319 men and 146 women, making a sample of about 7% all patients attending an STD clinic during the 3 month study period. Microbiological data were collected from 104 men and 72 women selected randomly from these. Pregnant women were excluded and patients who had received antibiotics within the previous 14 days were excluded from the microbiology sub-sample. SETTING--An outpatient STD clinic at a District Hospital on a major truck route about 300 km north of the capital, Harare. METHODS--All new patients attending the clinic during a 3 month period were enrolled for clinical and epidemiological investigations using a standard procedure. Specimens for microbiological investigation were taken from every second patient seen on the first three days of each week. RESULTS--The typical patient was male (m:f ratio 2.2) aged 20-29 years (68% patients), not married (56% men) and in paid employment (66% men vs. 27% for the district). In men the most common presenting feature was genital ulceration, while in women, discharges were more common. Genital warts were noted frequently in both sexes. In the sub-sample examined microbiologically, H ducreyi was isolated from 46% ulcers clinically diagnosed as chancroid, and motile spirochaetes were detected in 25% painless ulcers. Neither of these were detected in ulcers in women, but HSV antigen was found as frequently in ulcers from men (19%) as from women (17%). In patients with genital discharges, gonococcal infection occurred in 64% men and 17% women, while T vaginalis was isolated from 39% women and only 8% men. Over 60% gonococcal isolates were PPNG, and 18% showed in vitro resistance to tetracycline. Yeasts, mainly C albicans were isolated from 42% women with a discharge and 25% women with ulcers. In men the presence of yeasts was associated with superficial ulceration and itchiness of the glans. Positive HIV-1 serology was found in 64% patients. There was no statistical association with current genital ulcers, though there was an association with previous STD episodes and particularly with serological evidence of syphilis. Apart from yeasts, there was no association between positive HIV-1 serology and the presence of pathogens in the genital tract. CONCLUSIONS--The high prevalence of HIV-1 antibodies in STD patients in Karoi suggests integration of STD and AIDS control programmes to be a necessity. Since paid employment was a common feature of both STD clinic attendance and HIV-1 seropositivity, these programmes may be effectively directed through the work place.