title: Molecular characterization of mycobacteria isolated from seals.
abstract: Tuberculosis (TB) was diagnosed in 10 seals from three species (Arctocephalus australis, Arctocephalus tropicalis and Otaria flavescens) found in South America. The mycobacteria isolated from these cases belonged to the Mycobacterium tuberculosis complex, as determined by RFLP using an IS6110 probe, spoligotyping, analysis of the 16S rRNA gene sequence and by PCR-restriction analysis of hsp65. Polymorphisms in gyrA, katG, oxyR and pncA were investigated in some of the isolates, as well as the presence of the MPB70 antigen. The insertion sequence IS6110 was present in three to seven copies in the genome of the mycobacteria isolated from seals. Using the IS6110 probe, six patterns (designated A, B, C, D, E and F) were identified from 10 different isolates. Patterns A and B were found for the mycobacteria isolated from two and four seals, respectively, indicating an epidemiological relationship between isolates grouped according to their IS6110 RFLP. The mycobacteria isolated from seals shared the majority of their IS6110 DNA-containing restriction fragments, and nine isolates had an identical spoligotype; only one isolate showed a minor difference in its spoligotype. In addition, none of these spoligotypes were found in other M. tuberculosis complex strains. These results suggest that the isolates from seals constitute a unique group of closely related strains. The mycobacteria isolated from seals showed polymorphisms at gyrA codon 95 and katG codon 463, as do group 1 M. tuberculosis, and M. bovis. Group 1 mycobacteria are associated with cluster cases. The spoligotypes found in the mycobacteria isolated from seals lack spacers 39-43, as does M. bovis, but the MPB70 antigen, which is highly expressed in M. bovis and minimally expressed in M. tuberculosis, was not detected in these mycobacteria. The mycobacteria isolated from seals also showed oxyR and pncA polymorphisms specific to M. tuberculosis. In conclusion, the mycobacteria that cause TB in seals in the South-Western Atlantic are a related group, and based on the combination of genetic characteristics, belong to a unique genotypic group within the M. tuberculosis complex.
authors.affiliation: Instituto de Biotecnologı́a, CICV/INTA, Buenos Aires, Argentina1.
authors.forename: M J
authors.initials: MJ
authors.lastname: Zumárraga
authors.affiliation: Departamento de Micobacterias, DILAB SENASA, C. C. 77 Morón (1708), Buenos Aires, Argentina2.
authors.forename: A
authors.initials: A
authors.lastname: Bernardell
authors.affiliation: Fundación Mundo Marino, San Clemente del Tuyú, Argentina3.
authors.forename: R
authors.initials: R
authors.lastname: Bastida
authors.affiliation: Fundación Mundo Marino, San Clemente del Tuyú, Argentina3.
authors.forename: V
authors.initials: V
authors.lastname: Quse
authors.affiliation: Fundación Mundo Marino, San Clemente del Tuyú, Argentina3.
authors.forename: J
authors.initials: J
authors.lastname: Loureiro
authors.affiliation: Instituto de Biotecnologı́a, CICV/INTA, Buenos Aires, Argentina1.
authors.forename: A
authors.initials: A
authors.lastname: Cataldi
authors.affiliation: Instituto de Biotecnologı́a, CICV/INTA, Buenos Aires, Argentina1.
authors.forename: F
authors.initials: F
authors.lastname: Bigi
authors.affiliation: Instituto de Biotecnologı́a, CICV/INTA, Buenos Aires, Argentina1.
authors.forename: A
authors.initials: A
authors.lastname: Alito
authors.affiliation: Laboratorio de Tuberculosis de la Dirección de Laboratorios Veterinarios (DILAVE) 'Miguel C. Rubino', Montevideo, Uruguay4.
authors.forename: M
authors.initials: M
authors.lastname: Castro Ramos
authors.affiliation: Departamento de Microbiologı́a, Salud Pública y Medicina Preventiva, Facultad de Medicina, Universidad de Zaragoza, Zaragoza, Spain5.
authors.forename: S
authors.initials: S
authors.lastname: Samper
authors.affiliation: Departamento de Microbiologı́a, Salud Pública y Medicina Preventiva, Facultad de Medicina, Universidad de Zaragoza, Zaragoza, Spain5.
authors.forename: I
authors.initials: I
authors.lastname: Otal
authors.affiliation: Departamento de Microbiologı́a, Salud Pública y Medicina Preventiva, Facultad de Medicina, Universidad de Zaragoza, Zaragoza, Spain5.
authors.forename: C
authors.initials: C
authors.lastname: Martin
authors.affiliation: Instituto de Biotecnologı́a, CICV/INTA, Buenos Aires, Argentina1.
authors.forename: M I
authors.initials: MI
authors.lastname: Romano
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics; D001426:Bacterial Proteins; D015373:Bacterial Typing Techniques; D018834:Chaperonin 60; D018833:Chaperonins / Q000235:genetics; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000032:analysis; D006168:Guinea Pigs; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D015183:Restriction Mapping; D046048:Seals, Earless / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D013020:South America; D014376:Tuberculosis / Q000453:epidemiology / Q000662:veterinary*
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D018834:Chaperonin 60; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; C074119:heat-shock protein 65, Mycobacterium; D018833:Chaperonins