Thyroglossal Duct Cyst Infection Caused by Neisseria gonorrhoeae: An Unusual Complication of Pharyngeal Gonorrhea.

ABSTRACT: Neisseria gonorrhoeae is a human obligate pathogen whose clinical expression of disease ranges from localized genital infection to involvement of extragenital sites such as the conjunctiva and throat. We describe the second case of a thyroglossal duct abscess due to N. gonorrhoeae, an uncommon complication of pharyngeal gonococcal infection. The fortuitous occurrence in the same individual of these 2 conditions that both exhibit an occult clinical presentation likely accounts for rarity of this infection. We discuss the pertinent gonococcal and host factors that underlie the clinical manifestations of this infection. A particular focus is the fundamental role that the binding of the gonococcal opacity-associated protein to the ubiquitous human carcinoembryonic cell adhesion molecule plays in the pathogenesis of pharyngeal gonorrhea.

A case report of fatal disseminated Mycobacterium colombiense infection in a renal transplant recipient.

We report the first case of disseminated Mycobacterium colombiense infection in a solid organ transplant recipient. Co-infection with Cryptococcus neoformans led to fatal multisystem organ failure. We review the pathogen and host factors contributing to these opportunistic infections.

Oral immunization using HgbA in a recombinant chancroid vaccine delivered by attenuated Salmonella typhimurium SL3261 in the temperature-dependent rabbit model.

Chancroid, a sexually transmitted genital ulcer disease caused by the Gram-negative bacterium Haemophilus ducreyi, facilitates the acquisition and transmission of HIV. An effective vaccine against chancroid has not been developed. In this preliminary study, the gene encoding the H. ducreyi outer membrane hemoglobin receptor HgbA was cloned into the plasmid pTETnir15. The recombinant construct was introduced into the attenuated Salmonella typhimurium SL3261 strain and stable expression was induced in vitro under anaerobic conditions. The vaccine strain was delivered into the temperature-dependent rabbit model of chancroid by intragastric immunization as a single dose, or as three doses administered at two-weekly intervals. No specific antibody to HgbA was elicited after either dose schedule. Although the plasmid vector survived in vivo passage for up to 15 days following single oral challenge, HgbA expression was restricted to plasmid isolates recovered one day after immunization. Rabbits inoculated with the 3-dose booster regimen achieved no protective immunity from homologous challenge. These results emphasize that refinements in plasmid design to enhance a durable heterologous protein expression are necessary for the development of a live oral vaccine against chancroid.

Identification and characterization of a heme periplasmic-binding protein in Haemophilus ducreyi.

Haemophilus ducreyi, a gram-negative and heme-dependent bacterium, is the causative agent of chancroid, a genital ulcer sexually transmitted infection. Heme acquisition in H. ducreyi proceeds via a receptor mediated process in which the initial event involves binding of hemoglobin and heme to their cognate outer membrane proteins, HgbA and TdhA, respectively. Following this specific interaction, the fate of the periplasmic deposited heme is unclear. Using protein expression profiling of the H. ducreyi periplasmic proteome, a periplasmic-binding protein, termed hHbp, was identified whose expression was enhanced under heme-limited conditions. The gene encoding this protein was situated in a locus displaying genetic characteristics of an ABC transporter. The purified protein bound heme in a dose-dependent and saturable manner and this binding was specifically competitively inhibited by heme. The hhbp gene functionally complemented an Escherichia coli heme uptake mutant. Expression of the heme periplasmic-binding protein was detected in a limited survey of H. ducreyi and H. influenzae clinical strains. These results indicate that the passage of heme into the cytoplasm of H. ducreyi involves a heme dedicated ABC transporter.

Use of signature-tagged mutagenesis to identify virulence determinants in Haemophilus ducreyi responsible for ulcer formation.

Elucidating the molecular mechanisms responsible for chancroid, a genital ulcer disease caused by Haemophilus ducreyi, has been hampered in part by the relative genetic intractability of the organism. A whole genome screen using signature-tagged mutagenesis in the temperature-dependent rabbit model (TDRM) of H. ducreyi infection uncovered 26 mutants with a presumptive attenuated phenotype. Insertions in two previously recognized virulence determinants, hgbA and lspA1, validated this genome scanning technique. Database interrogation allowed assignment of 24 mutants to several functional classes, including transport, metabolism, DNA repair, stress response and gene regulation. The attenuated virulence for a 3 strain with a mutation in hicB was confirmed by individual infection in the TDRM. The results from this preliminary study indicate that this high throughput strategy will further the understanding of the pathogenesis of H. ducreyi infection.

Trichomonas vaginalis: the adhesins AP51 and AP65 bind heme and hemoglobin.

Trichomonas vaginalis is the cause of human trichomoniasis, the most common non-viral sexually transmitted disease worldwide. Although acquisition of iron by binding to host hemoglobin through distinct receptor(s) has been described, no specific heme- or hemoglobin-binding site has been reported in this parasite. To determine the presence of hemoglobin-binding protein(s), membrane proteins were subjected to hemoglobin-affinity chromatography. Eluted proteins were analysed by SDS-PAGE. Two protein bands of 48 and 63 kDa were detected. Competition assay with an excess amount of hemoglobin or hemin in hemoglobin-affinity chromatography could block the 63- and 48-kDa bands, respectively. Further analysis by mass spectrometry indicated that the 48- and 63-kDa proteins had identity with two T. vaginalis adhesins: AP51 and AP65, respectively. This study confirms the existence of multifunctional proteins in T. vaginalis, and suggested that AP51 and AP65, besides serving as adhesion molecules, could also act as heme- and hemoglobin-binding proteins.

Successful treatment of a prosthetic joint infection due to Mycobacterium abscessus.

Prosthetic joint infection due to Mycobacterium abscessus is uncommon and optimal therapy remains poorly defined. Following a two-stage revision, clinical and microbiological cure was achieved in a patient with a M abscessus-infected total hip arthroplasty. A prolonged course of directed antibacterial therapy comprising clarithromycin and cefoxitin coupled with the application of amikacin-impregnated cement likely contributed to the successful outcome.

Epidemiology of infectious syphilis in Ottawa. Recurring themes revisited.

OBJECTIVES: To describe the epidemiology of an outbreak of infectious syphilis in Ottawa. METHODS: A retrospective chart review of infectious syphilis cases in Ottawa from 2001-2006. RESULTS: Rates of syphilis have risen more than tenfold. The epidemic was centered in men, with the majority of cases (83.5%) occurring among men who have sex with men (MSM). These individuals differed from the general MSM population residing in Ottawa in their being older, more likely to be HIV positive, and more sexually promiscuous. Inconsistent condom use by MSM engaged in either oral or anal sex was pervasive. Thirty-seven percent of MSM reported sexual encounters with men from Montreal and Toronto. Visceral manifestations of syphilis, including neurosyphilis, were more common in persons co-infected with HIV. As a result, this subgroup was more likely to have received an extended antibiotic treatment regimen. There was a substantial delay between serological diagnosis and treatment. Less than half of treated cases returned for a six-month evaluation. CONCLUSIONS: Multiple sexual partners, unprotected oral sex, and increased age among MSM were the predominant risk factors contributing to this syphilis epidemic. Co-infection with HIV modified the clinical presentation of syphilis, necessitating a more intensive diagnostic and therapeutic approach. The interconnection of urban sexual networks has likely contributed to the dynamics of local syphilis transmission and suggests that effective interventions will require a coordinated national approach.

Central venous catheter-associated Leifsonia aquatica bacteremia in a hemodialysis-dependent patient.

Infections associated with Leifsonia aquatica are particularly uncommon. We describe a central venous catheter-associated L. aquatica bacteremia in a hemodialysis-dependent patient. A review of the literature revealed only 1 other case report involving 10 hemodialysis patients with documented L. aquatica bacteremia.

A role for Haemophilus ducreyi Cu,ZnSOD in resistance to heme toxicity.

The Cu,Zn superoxide dismutase (Cu,ZnSOD) from Haemophilus ducreyi is the only enzyme of this class which binds a heme molecule at its dimer interface. To explore the role of the enzyme in this heme-obligate bacterium, a sodC mutant was created by insertional inactivation. No difference in growth rate was observed during heme limitation. In contrast, under heme rich conditions growth of the sodC mutant was impaired compared to the wild type strain. This growth defect was abolished by supplementation of exogenous catalase. Genetic complementation of the sodC mutant in trans demonstrated that the enzymatic property or the heme-binding activity of the protein could repair the growth defect of the sodC mutant. These results indicate that Cu,ZnSOD protects Haemophilus ducreyi from heme toxicity.

Attenuated Salmonella typhimurium SL3261 as a vaccine vector for recombinant antigen in rabbits.

Oral live Salmonella vaccine vectors expressing recombinant guest antigens help stimulate systemic, mucosal, humoral, and cell-mediated immune responses against Salmonella and recombinant antigens. It may be possible to use them effectively against Haemophilus ducreyi, the bacterium that causes chancroid, a sexually transmitted genital ulcer disease. This study aimed to test the feasibility of using oral Salmonella vaccine vectors for the evaluation of chancroid vaccine candidates in the temperature-dependent rabbit model of H. ducreyi infection, an in vivo quantitative virulence assay of inducible immunity. We identified 10(8) to 10(9) CFU to be a safe and immunogenic oral dose range of S. typhimurium SL3261, by monitoring post-administration onset and course of illness and antibody titre by enzyme immunoassay (EIA). We successfully transduced plasmid pTETnir15 into the strain to produce recombinant S. typhimurium SL3261(pTETnir15), successfully expressed tetanus toxin fragment C (TetC) in it, and elicited serum anti-TetC titres of 1:6400 by EIA, 4 weeks after inoculation. The course of experimentally induced H. ducreyi skin lesions in rabbits treated with SL3261(pTETnir15) was similar to that in saline-treated controls. We describe a framework that successfully uses Salmonella as a vector for recombinant control antigen in the rabbit model of H. ducreyi infection, and is suitable for pre-clinical evaluation of Salmonella vector-based H. ducreyi vaccine antigen candidates.

Exploring the pathogenesis of necrotizing fasciitis due to Streptococcus pneumoniae.

Monobacterial necrotizing fasciitis is a rare form of soft tissue infection usually caused by the group A beta-hemolytic Streptococcus. Soft tissue infection is an uncommon clinical manifestation of invasive disease due to Streptococcus pneumoniae. We describe 3 cases of pneumococcal necrotizing fasciitis and explore potential pathogen-specific mechanisms of pathogenesis. The clinical characteristics of necrotizing fasciitis due to S. pneumoniae and group A beta-hemolytic Streptococcus appear to overlap. The similarities include predominant occurrence in elderly adults with underlying chronic illness, predilection for lower extremity infection, progression to toxic shock-like syndrome and a high case fatality rate. No DNA fragments corresponding to speA, speB or speC were amplified by PCR from the 3 pneumococcal isolates. Western immunoblot revealed no evidence of SpeA, SpeB or SpeC protein expression. Evaluation for protease production and cytotoxicity was unrevealing. The similar clinical presentation of pneumococcal necrotizing fasciitis to the disease caused by the group A beta-hemolytic Streptococcus has important therapeutic implications. The molecular mechanisms underlying the pathogenesis are unclear. Prospective population-based studies are required to define the epidemiology of this infection.