@article {12, title = {First report of a mycolactone-producing Mycobacterium infection in fish agriculture in Belgium.}, journal = {FEMS Microbiol Lett}, volume = {286}, year = {2008}, month = {2008 Sep}, pages = {93-5}, abstract = {

In the past few years, a mycolactone-producing subgroup of the Mycobacterium marinum complex has been identified and analyzed. These IS2404-positive species cause pathology in frogs and fish. A recently isolated mycobacterial strain from a fish in Belgium was analyzed using a variety of molecular methods and the results were identical to those obtained from a mycolactone-producing M. marinum from Israel.

}, keywords = {Agriculture, Animals, Bacterial Proteins, Bacterial Toxins, Belgium, Fish Diseases, Fishes, Israel, Macrolides, Mycobacterium Infections, Nontuberculous, Mycobacterium marinum}, issn = {0378-1097}, doi = {10.1111/j.1574-6968.2008.01264.x}, author = {Stragier, Pieter and Hermans, Katleen and Stinear, Tim and Portaels, Fran{\c c}oise} } @article {13, title = {A comparison of DNA extraction procedures for the detection of Mycobacterium ulcerans, the causative agent of Buruli ulcer, in clinical and environmental specimens.}, journal = {J Microbiol Methods}, volume = {76}, year = {2009}, month = {2009 Feb}, pages = {152-8}, abstract = {

Mycobacterium ulcerans is the causative agent of Buruli ulcer, the third most common mycobacterial disease in humans after tuberculosis and leprosy. Although the disease is associated with aquatic ecosystems, cultivation of the bacillus from the environment is difficult to achieve. Therefore, at the moment, research is based on the detection by PCR of the insertion sequence IS2404 present in M. ulcerans and some closely related mycobacteria. In the present study, we compared four DNA extraction methods for detection of M. ulcerans DNA, namely the one tube cell lysis and DNA extraction procedure (OT), the FastPrep procedure (FP), the modified Boom procedure (MB), and the Maxwell 16 Procedure (M16). The methods were performed on serial dilutions of M. ulcerans, followed by PCR analysis with different PCR targets in M. ulcerans to determine the detection limit (DL) of each method. The purity of the extracted DNA and the time and effort needed were compared as well. All methods were performed on environmental specimens and the two best methods (MB and M16) were tested on clinical specimens for detection of M. ulcerans DNA. When comparing the DLs of the DNA extraction methods, the MB and M16 had a significantly lower DL than the OT and FP. For the different PCR targets, IS2404 showed a significantly lower DL than mlsA, MIRU1, MIRU5 and VNTR6. The FP and M16 were considerably faster than the MB and OT, while the purity of the DNA extracted with the MB was significantly higher than the DNA extracted with the other methods. The MB performed best on the environmental and clinical specimens. This comparative study shows that the modified Boom procedure, although lengthy, provides a better method of DNA extraction than the other methods tested for detection and identification of M. ulcerans in both clinical and environmental specimens.

}, keywords = {Animals, Buruli Ulcer, DNA Transposable Elements, DNA, Bacterial, DNA, Plant, Environmental Monitoring, Humans, Mycobacterium ulcerans, Polymerase Chain Reaction, Reagent Kits, Diagnostic, Sensitivity and Specificity, Soil Microbiology, Water Microbiology}, issn = {0167-7012}, doi = {10.1016/j.mimet.2008.10.002}, author = {Durnez, Lies and Stragier, Pieter and Roebben, Karen and Ablordey, Anthony and Leirs, Herwig and Portaels, Fran{\c c}oise} } @article {6, title = {Evidence for an intramacrophage growth phase of Mycobacterium ulcerans.}, journal = {Infect Immun}, volume = {75}, year = {2007}, month = {2007 Feb}, pages = {977-87}, abstract = {

Mycobacterium ulcerans is the etiologic agent of Buruli ulcer (BU), an emerging tropical skin disease. Virulent M. ulcerans secretes mycolactone, a cytotoxic exotoxin with a key pathogenic role. M. ulcerans in biopsy specimens has been described as an extracellular bacillus. In vitro assays have suggested a mycolactone-induced inhibition of M. ulcerans uptake by macrophages in which its proliferation has not been demonstrated. Therefore, and uniquely for a mycobacterium, M. ulcerans has been classified as an extracellular pathogen. In specimens from patients and in mouse footpad lesions, extracellular bacilli were concentrated in central necrotic acellular areas; however, we found bacilli within macrophages in surrounding inflammatory infiltrates. We demonstrated that mycolactone-producing M. ulcerans isolates are efficiently phagocytosed by murine macrophages, indicating that the extracellular location of M. ulcerans is not a result of inhibition of phagocytosis. Additionally, we found that M. ulcerans multiplies inside cultured mouse macrophages when low multiplicities of infection are used to prevent early mycolactone-associated cytotoxicity. Following the proliferation phase within macrophages, M. ulcerans induces the lysis of the infected host cells, becoming extracellular. Our data show that M. ulcerans, like M. tuberculosis, is an intracellular parasite with phases of intramacrophage and extracellular multiplication. The occurrence of an intramacrophage phase is in accordance with the development of cell-mediated and delayed-type hypersensitivity responses in BU patients.

}, keywords = {Animals, Bacterial Toxins, Cells, Cultured, Disease Models, Animal, Female, Foot, Histocytochemistry, Humans, Macrolides, Macrophages, Mice, Mice, Inbred BALB C, Microscopy, Electron, Transmission, Mycobacterium Infections, Nontuberculous, Mycobacterium ulcerans, Phagocytosis, Skin Diseases, Bacterial, Skin Ulcer}, issn = {0019-9567}, doi = {10.1128/IAI.00889-06}, author = {Torrado, Eg{\'\i}dio and Fraga, Alexandra G and Castro, Ant{\'o}nio G and Stragier, Pieter and Meyers, Wayne M and Portaels, Fran{\c c}oise and Silva, Manuel T and Pedrosa, Jorge} } @article {7, title = {Mycobacterium liflandii infection in European colony of Silurana tropicalis.}, journal = {Emerg Infect Dis}, volume = {13}, year = {2007}, month = {2007 May}, pages = {743-6}, abstract = {

Mycobacterium liflandii causes a fatal frog disease in captive anurans. Here we report, to our knowledge, the first epizootic of mycobacteriosis in a European colony of clawed frogs (Silurana tropicalis), previously imported from a United States biologic supply company. Our findings suggest the emerging potential of this infection through international trade.

}, keywords = {Animals, Europe, Models, Animal, Mycobacterium, Mycobacterium Infections, United States, Xenopus, Zoonoses}, issn = {1080-6040}, doi = {10.3201/eid1305.060625}, author = {Suykerbuyk, Patrick and Vleminckx, Kris and Pasmans, Frank and Stragier, Pieter and Ablordey, Anthony and Tran, Hong Thi and Hermans, Katleen and Fleetwood, Michelle and Meyers, Wayne M and Portaels, Fran{\c c}oise} } @article {10, title = {First cultivation and characterization of Mycobacterium ulcerans from the environment.}, journal = {PLoS Negl Trop Dis}, volume = {2}, year = {2008}, month = {2008}, pages = {e178}, abstract = {

BACKGROUND: Mycobacterium ulcerans disease, or Buruli ulcer (BU), is an indolent, necrotizing infection of skin, subcutaneous tissue and, occasionally, bones. It is the third most common human mycobacteriosis worldwide, after tuberculosis and leprosy. There is evidence that M. ulcerans is an environmental pathogen transmitted to humans from aquatic niches; however, well-characterized pure cultures of M. ulcerans from the environment have never been reported. Here we present details of the isolation and characterization of an M. ulcerans strain (00-1441) obtained from an aquatic Hemiptera (common name Water Strider, Gerris sp.) from Benin.

METHODOLOGY/PRINCIPAL FINDINGS: One culture from a homogenate of a Gerris sp. in BACTEC became positive for IS2404, an insertion sequence with more than 200 copies in M. ulcerans. A pure culture of M. ulcerans 00-1441 was obtained on L{\"o}wenstein-Jensen medium after inoculation of BACTEC culture in mouse footpads followed by two other mouse footpad passages. The phenotypic characteristics of 00-1441 were identical to those of African M. ulcerans, including production of mycolactone A/B. The nucleotide sequence of the 5{\textquoteright} end of 16S rRNA gene of 00-1441 was 100\% identical to M. ulcerans and M. marinum, and the sequence of the 3{\textquoteright} end was identical to that of the African type except for a single nucleotide substitution at position 1317. This mutation in M. ulcerans was recently discovered in BU patients living in the same geographic area. Various genotyping methods confirmed that strain 00-1441 has a profile identical to that of the predominant African type. Strain 00-1441 produced severe progressive infection and disease in mouse footpads with involvement of bone.

CONCLUSION: Strain 00-1441 represents the first genetically and phenotypically identified strain of M. ulcerans isolated in pure culture from the environment. This isolation supports the concept that the agent of BU is a human pathogen with an environmental niche.

}, keywords = {Animals, Bacterial Toxins, Cells, Cultured, Environmental Microbiology, Female, Foot, Genotype, Hemiptera, Macrolides, Macrophages, Mass Spectrometry, Mice, Mice, Inbred BALB C, Mycobacterium ulcerans, Phylogeny, Polymerase Chain Reaction, RNA, Ribosomal, 16S}, issn = {1935-2735}, doi = {10.1371/journal.pntd.0000178}, author = {Portaels, Fran{\c c}oise and Meyers, Wayne M and Ablordey, Anthony and Castro, Ant{\'o}nio G and Chemlal, Karim and de Rijk, Pim and Elsen, Pierre and Fissette, Krista and Fraga, Alexandra G and Lee, Richard and Mahrous, Engy and Small, Pamela L C and Stragier, Pieter and Torrado, Eg{\'\i}dio and Van Aerde, Anita and Silva, Manuel T and Pedrosa, Jorge} } @article {1, title = {Genetic diversity and population structure of Mycobacterium marinum: new insights into host and environmental specificities.}, journal = {J Clin Microbiol}, volume = {50}, year = {2012}, pages = {3627-34}, abstract = {

Mycobacterium marinum causes a systemic tuberculosis-like disease in fish and skin infections in humans that can spread to deeper structures, resulting in tenosynovitis, arthritis, and osteomyelitis. However, little information is available concerning (i) the intraspecific genetic diversity of M. marinum isolated from humans and animals; (ii) M. marinum genotype circulation in the different ecosystems, and (iii) the link between M. marinum genetic diversity and hosts (humans and fish). Here, we conducted a genetic study on 89 M. marinum isolates from humans (n = 68) and fish (n = 21) by using mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing. The results show that the M. marinum population is genetically structured not only according to the host but also according to the ecosystem as well as to tissue tropism in humans. This suggests the existence of different genetic pools in the function of the biological and ecological compartments. Moreover, the presence of only certain M. marinum genotypes in humans suggests a different zoonotic potential of the M. marinum genotypes. Considering that the infection is linked to aquarium activity, a significant genetic difference was also detected when the human tissue tropism of M. marinum was taken into consideration, with a higher genetic polymorphism in strains isolated from patients with cutaneous forms than from individuals with deeper-structure infection. It appears that only few genotypes can produce deeper infections in humans, suggesting that the immune system might play a filtering role.

}, keywords = {Adolescent, Adult, Aged, Animals, Biota, Child, Child, Preschool, DNA, Bacterial, Female, Fish Diseases, Fishes, Genetic Variation, Genotype, Humans, Interspersed Repetitive Sequences, Male, Middle Aged, Molecular Typing, Mycobacterium Infections, Nontuberculous, Mycobacterium marinum, Young Adult}, issn = {1098-660X}, doi = {10.1128/JCM.01274-12}, author = {Broutin, Vincent and Ba{\~n}uls, Anne-Laure and Aubry, Alexandra and Keck, Nicolas and Choisy, Marc and Bernardet, Jean-Fran{\c c}ois and Michel, Christian and Raymond, Jean-Christophe and Libert, C{\'e}dric and Barnaud, Antoine and Stragier, Pieter and Portaels, Fran{\c c}oise and Terru, Dominique and Belon, Claudine and Dereure, Olivier and Gutierrez, Cristina and Boschiroli, Maria-Laura and Van De Perre, Philippe and Cambau, Emmanuelle and Godreuil, Sylvain} }