John A. Hayman

Buruli Ulcer (M. ulcerans Infection): New Insights, New Hope for Disease Control

Buruli ulcer is a disease of skin and soft tissue caused by Mycobacterium ulcerans. It can leave affected people scarred and disabled. What are the prospects for disease control?

Mycobacterium ulcerans in Mosquitoes Captured during Outbreak of Buruli Ulcer, Southeastern Australia

Mosquitoes positive for M. ulcerans were linked to outbreaks of Buruli ulcer in humans.

Identification of Mycobacterium ulcerans in the Environment from Regions in Southeast Australia in Which It Is Endemic with Sequence Capture-PCR

ABSTRACT We recently described the use of PCR to identify the environmental source of Mycobacterium ulcerans during an outbreak of ulcerative disease that occurred in a localized region of southeast Australia. The PCR used was based on amplification of the M. ulcerans-specific insertion sequence, IS2404. In this study we developed a new test that is a substantial improvement over the original PCR method in terms of sensitivity, reliability, and ease of use. In the new method magnetic bead sequence capture-PCR is used to detect two M. ulcerans sequences (IS2404 and IS2606) and total mycobacterial 16S ribosomal DNA. We used sequence capture-PCR to test water and plant material collected over a 12-month period during 1998 and 1999 from sites near the centers of two distinct foci of M. ulcerans infections. A golf course irrigation system in one area and a small shallow lake in another area repeatedly were PCR positive for M. ulcerans. Nearby sites and sites unrelated to the endemic areas were negative. Based on the PCR data, a most-probable-number method was used to estimate the concentration of M. ulcerans cells in positive samples from both regions. This procedure resulted in average concentrations of 0.5 cell per 100 ml of water and 40 cells per 100 g of detritus. Loss of the PCR signal coincided with a decrease in ulcerative disease in each area. These results provide further evidence that M. ulcerans may be transmitted from a point environmental source and demonstrate the utility of magnetic bead sequence capture-PCR for identification of nonculturable microbial pathogens in the environment.

Mycobacterium ulcerans toxic macrolide, mycolactone modulates the host immune response and cellular location of M. ulcerans in vitro and in vivo

Mycobacterium ulcerans produces an extracellular cutaneous infection (Buruli ulcer) characterized by immunosuppression. This is in stark contrast to all other pathogenic Mycobacteria species that cause intracellular, granulomatous infections. The unique mycobacterial pathology of M. ulcerans infection is attributed to a plasmid‐encoded immunomodulatory macrolide toxin, mycolactone. In this article we explore the role of mycolactone in the virulence of M. ulcerans using mycolactone and genetically defined mycolactone negative mutants. In a guinea pig infection model wild‐type (WT) M. ulcerans produces an extracellular infection whereas mycolactone negative mutants produce an intracellular inflammatory infection similar to that of Mycobacterium marinum. Although mycolactone negative mutants are avirulent, they persist for at least 6 weeks. Chemical complementation of M. ulcerans mutants with mycolactone restores WT M. ulcerans pathology. Mycolactone negative mutants are capable of growth within macrophages in vitro whereas macrophages are killed by WT M. ulcerans. The ability of mycolactone to caused delayed cell death via apoptosis has been reported. However, mycolactone also causes cell death via necrosis. In vitro mycolactone has antiphagocytic properties. Neither WT M. ulcerans nor mycolactone negative strains are strong neutrophil attractants. These results suggest that mycolactone is largely responsible for the unique pathology produced by M. ulcerans.

Immune Response to Infection with Mycobacterium ulcerans

ABSTRACT Mycobacterium ulcerans is a slow-growing, acid-fast bacillus that causes chronic necrotizing skin ulcers known as Buruli ulcers. Previously reported information on immunity to this mycobacterium is limited. We examined immune responses to M. ulcerans and M. bovis BCG in patients with M. ulcerans disease and in 20 healthy control subjects (10 tuberculin test positive and 10 tuberculin test negative). Cell-mediated immunity was assessed by stimulating peripheral blood mononuclear cells (PBMC) with whole mycobacteria and then measuring PBMC proliferation and the production of gamma interferon (IFN-γ). Humoral immunity was assessed by immunoblotting. PBMC from all subjects showed significantly greater proliferation and IFN-γ production in response to stimulation with living mycobacteria compared with killed cells. However, PBMC from subjects with past or current M. ulcerans disease showed significantly reduced proliferation and production of IFN-γ in response to stimulation with live M. ulcerans or M. bovis than PBMC from healthy, tuberculin test-positive subjects (P < 0.001) and showed results in these assays comparable to those of tuberculin test-negative subjects (P > 0.2). Serum from 9 of 11 patients with M. ulcerans disease, but no control subject, contained antibodies to M. ulcerans. The results indicate that patients with M. ulcerans infection mount an immune response to M. ulcerans as evidenced by antibody production, but they demonstrate profound systemic T-cell anergy to mycobacterial antigens. These findings may explain some of the distinct clinical and pathological features of M. ulcerans-induced disease.

Localised Mycobacterium ulcerans infection in a cat in Australia

A 10-year-old castrated male domestic cat domiciled in eastern Victoria (Australia) was presented for a subcutaneous mass on its nasal bridge in November 2006. Cytological examination of an aspirate demonstrated pyogranulomatous inflammation. At surgery, the lesion consisted of an encapsulated mass containing viscid fluid. Histological examination of the resected lesion revealed pyogranulomatous inflammation surrounding a central zone of necrosis. Sections stained with the Ziehl–Neelsen method revealed numerous acid-fast bacilli, intracellularly within macrophages and extracellularly. Molecular studies established the infection was caused by Mycobacterium ulcerans. As histology demonstrated that the infection extended to the margin of the excised tissues, the cat was treated subsequently with clarithromycin (62.5 mg orally once daily for 7 days, then twice daily for 3 months). The surgical wound healed unremarkably. The infection has not recurred at the time of writing, 1 year following discontinuation of treatment. Although M ulcerans infections have been recorded in variety of mammals, this is the first known case in a cat.

All-Oral Antibiotic Treatment for Buruli Ulcer: A Report of Four Patients

1Department of Infectious Diseases, Austin Health, Melbourne, Australia, 2Department of Surgery, Box Hill Hospital, Melbourne, Australia, 3Department of Surgery,Monash University, Melbourne, Australia, 4Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia, 5WHO Collaborating Centrefor Mycobacterium ulcerans (Western Pacific Region) and Victorian Infectious Diseases Reference Laboratory, Melbourne, Australia, 6Department of AnatomicalPathology, Box Hill Hospital, Melbourne, Australia, 7Department of Infectious Diseases, Royal Children’s Hospital, Melbourne, Australia

Climate and Landscape Factors Associated with Buruli Ulcer Incidence in Victoria, Australia

Background Buruli ulcer (BU), caused by Mycobacterium ulcerans (M. ulcerans), is a necrotizing skin disease found in more than 30 countries worldwide. BU incidence is highest in West Africa; however, cases have substantially increased in coastal regions of southern Australia over the past 30 years. Although the mode of transmission remains uncertain, the spatial pattern of BU emergence in recent years seems to suggest that there is an environmental niche for M. ulcerans and BU prevalence. Methodology/Principal Findings Network analysis was applied to BU cases in Victoria, Australia, from 1981–2008. Results revealed a non-random spatio-temporal pattern at the regional scale as well as a stable and efficient BU disease network, indicating that deterministic factors influence the occurrence of this disease. Monthly BU incidence reported by locality was analyzed with landscape and climate data using a multilevel Poisson regression approach. The results suggest the highest BU risk areas occur at low elevations with forested land cover, similar to previous studies of BU risk in West Africa. Additionally, climate conditions as far as 1.5 years in advance appear to impact disease incidence. Warmer and wetter conditions 18–19 months prior to case emergence, followed by a dry period approximately 5 months prior to case emergence seem to favor the occurrence of BU. Conclusions/Significance The BU network structure in Victoria, Australia, suggests external environmental factors favor M. ulcerans transmission and, therefore, BU incidence. A unique combination of environmental conditions, including land cover type, temperature and a wet-dry sequence, may produce habitat characteristics that support M. ulcerans transmission and BU prevalence. These findings imply that future BU research efforts on transmission mechanisms should focus on potential vectors/reservoirs found in those environmental niches. Further, this study is the first to quantitatively estimate environmental lag times associated with BU outbreaks, providing insights for future transmission investigations.

Spontaneous Clearance of Mycobacterium ulcerans in a Case of Buruli Ulcer

Buruli ulcer (BU) is an infection of skin and soft tissue caused by Mycobacterium ulcerans, a toxin-producing environmental mycobacterium. Significant advances in the treatment of BU have been made over the past decade with the introduction of effective antibiotic therapy and there is a greater understanding of the pathogenesis and host immune response. Although it is generally held that early BU lesions may heal spontaneously, to our knowledge, there are no previously published cases that definitively document spontaneous resolution of culture-confirmed BU.

Mycobacterium ulcerans infection diagnosed by polymerase chain reaction

Abstract:  Mycobacterium ulcerans infection is the third most important mycobacterial infection world‐wide affecting immunocompetent individuals and causes chronic progressive skin ulcers. It has been described in many different regions world‐wide. The diagnosis of M. ulcerans infection is often delayed because the diagnosis is difficult to make when new cases appear outside known endemic areas. However, molecular methods are now available to diagnose and distinguish M. ulcerans from other mycobacteria, allowing rapid diagnosis. Presented here is the case of a previously well girl from Townsville, Queensland, with extensive M. ulcerans infection involving the elbow joint, triceps tendon and underlying bone. Rapid diagnosis by polymerase chain reaction confirmed M. ulcerans infection. This is the first known case of M. ulcerans infection from Townsville in over 25 years, highlighting the changing epidemiology of this disease.