We describe the first transmission of Mycobacterium tuberculosis from human to cattle confirmed by molecular typing of isolates involved in the transmission. IS6110-based restriction fragment length polymorphism analysis showed that the isolates from the cattle and farm worker who suffered from pulmonary tuberculosis 1 year prior to this case were the same strains.

BackgroundSince non-tuberculous mycobacteria (NTM) disease is not notifiable in most European Union (EU) and European Economic Area (EEA) countries, the epidemiological situation of the >150 NTM species is largely unknown. We aimed to collect data on the frequency of NTM detection and NTM species types in EU/EEA countries.MethodsOfficially nominated national tuberculosis reference laboratories of all EU/EEA countries were asked to provide information on: laboratory routines for detection and identification of NTM, including drug sensitivity testing (DST) methods; data on the number and type of NTM species identified; coverage and completeness of the provided data on NTM; type and number of human specimens tested for NTM; and number of specimens tested for Mycobacterium tuberculosis complex and NTM. This information was summarized and the main results are described.ResultsIn total, 99 different NTM species were identified with M. avium, M. gordonae, M. xenopi , M. intracellulare, and M. fortuitum identified most frequently. Seven percent of the NTM species could not be identified. NTM was cultured from between 0.4-2.0% of the specimens (data from four countries). The laboratories use culturing methods optimised for M. tuberculosis complex. Identification is mainly carried out by a commercial line probe assay supplemented with sequencing. Most laboratories carried out DST for rapid growers and only at the explicit clinical request for slow growers.ConclusionIt is likely that the prevalence of NTM is underestimated because diagnostic procedures are not optimized specifically for NTM and isolates may not be referred to the national reference laboratory for identification. Due to the diagnostic challenges and the need to establish the clinical relevance of NTM, we recommend that countries should concentrate detection and identification in only few laboratories.

To study the dissemination of Mycobacterium bovis subsp. caprae, 79 European isolates from cattle, humans, and other hosts were examined by spoligotyping and IS6110 restriction fragment length polymorphism (RFLP) analysis. Among a total of 11 different spoligotypes identified, type C1 proved to be predominant (n ‫؍‬ 62). Five of the spoligotypes are described for the first time. A total of 43 different RFLP types were identified, thus allowing further differentiation for epidemiological tracking. Isolates from a series of outbreaks in one village proved to be of the same spoligotype and of identical or closely related RFLP types. Mycobacterium bovis subsp. caprae is a member of the M. tuberculosis complex, which also includes M. tuberculosis, M. africanum, M. bovis, M. bovis BCG, M. microti, and M. canettii.General interest in this pathogen, which was isolated from tuberculous lesions in humans, cattle, goats, sheep, and deer, has been steadily increasing since its first description (3). Thus, the prevalence of M. bovis subsp. caprae was extensively studied in Spain (4, 7) and France (8). There have been several reports about its occurrence in animals and humans in Austria (18) and the Czech Republic (15,16,17) and also in Germany (13), where it is reported to account for about one-third of human M. bovis-associated cases of tuberculosis (10).Identification of the various M. tuberculosis complex organisms by spoligotyping (9, 12) is performed on the basis of the presence or absence of certain combinations of 43 spacers which are interspersed among a high number of conserved repeats in the chromosomal direct-repeat region. Additional intraspecies genetic differences can be revealed by IS6110 fingerprinting or IS6110 restriction fragment length polymorphism (RFLP) analysis (19), which takes advantage of the mobility of insertion element IS6110 along the mycobacterial chromosome.The combination of spoligotyping and IS6100 RFLP can be expected to improve the possibilities of revealing epidemiological relationships because of the different discriminatory potential of both methods (18). Recent studies of serial isolates of M. tuberculosis clearly showed that due to the relatively frequent occurrence of IS6110 transposition events, IS6100 RFLP patterns were more likely to change within a few years of a strain's appearance than spoligotyping patterns (1,6,20).

Thirty-five aquarium fish were investigated for the presence of mycobacteria by culture and molecular methods. The following species were examined: goldfish Carassius auratus auratus, guppy Poecilia reticulata, 4 three-spot gourami Trichogaster trichopterus, dwarf gourami Colisa lalia, Siamese fighting fish Betta splendens, freshwater angelfish Pterophyllum scalare, African cichlid fish Cichlidae spp., cichlid fish Microgeophagus altispinosus, cichlid fish Pseudotropheus lombardoi, blue streak hap Labidochromis caeruleus, sterlet Acipenser ruthenus, southern platyfish Xiphophorus maculatus, and catfish Corydoras spp. Isolates of mycobacteria were obtained in 29 cases (82.9%). Two specimens were positive using Ziehl-Neelsen (ZN) staining, but the cultivation failed. Four specimens were both ZN-and culture-negative. On the basis of GenoType Mycobacterium assay (Hain Lifescience) and restriction enzyme analysis of the amplified products (PCR-RFLP), 23 isolates (79.3%) were identified: 7 as Mycobacterium fortuitum, 6 as M. gordonae, 6 as M. marinum, 3 as M. chelonae, and 1 as M. peregrinum. Five isolates remained unidentified (Mycobacterium spp.). One case probably represented a mixed infection (M. marinum/M. fortuitum). Since M. marinum infections are also detected in humans, the significance of mycobacteria in aquarium fish should not be overlooked. KEY WORDS: Mycobacteria · Aquarium fish · PCR · Restriction enzyme analysis Resale or republication not permitted without written consent of the publisherDis Aquat Org 64: [29][30][31][32][33][34][35] 2005 character of the disease, any suspicion of fish mycobacteriosis should be taken seriously and investigation of the case conducted.The diagnosis of mycobacterial disease in fish is based on histopathological, culture and molecular methods. The aim of the present study was to identify the species of mycobacteria isolated from aquarium fish in Slovenia in the period from 2001 to 2004 and to compare the available identification methods in order to assess their suitability for routine use. MATERIALS AND METHODSMaterials. The internal organs of 35 aquarium fish were investigated. The number of fish and the species investigated were: 11 goldfish Carassius auratus auratus, 7 freshwater angelfish Pterophyllum scalare, 4 three-spot gourami Trichogaster trichopterus, 3 guppy Poecilia reticulata, 2 southern platyfish Xiphophorus maculatus, 1 dwarf gourami Colisa lalia, 1 Siamese fighting fish Betta splendens, 1 African cichlid fish Cichlidae spp., 1 cichlid fish Microgeophagus altispinosus, 1 cichlid fish Pseudotropheus lombardoi, 1 blue streak hap Labidochromis caeruleus, 1 sterlet Acipenser ruthenus, and 1 catfish Corydoras spp.Bacteriology. Smears made directly from fish organs were stained with Ziehl-Neelsen (ZN). Following homogenization, decontamination with N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH), and concentration of the specimens, their sediments were inoculated on the following media: Stonebrink, Middlebrook 7H10, Löwenstein-Jensen with pyruvate, ...

Only very recently, has it been proposed that the hitherto existing Mycobacterium kansasii subtypes (I-VI) should be elevated, each, to a species rank. Consequently, the former M. kansasii subtypes have been denominated as Mycobacterium kansasii (former type I), Mycobacterium persicum (II), Mycobacterium pseudokansasii (III), Mycobacterium innocens (V), and Mycobacterium attenuatum (VI). The present work extends the recently published findings by using a three-pronged computational strategy, based on the alignment fraction-average nucleotide identity, genome-to-genome distance, and core-genome phylogeny, yet essentially independent and much larger sample, and thus delivers a more refined and complete picture of the M. kansasii complex. Furthermore, five canonical taxonomic markers were used, i.e., 16S rRNA, hsp65, rpoB, and tuf genes, as well as the 16S-23S rRNA intergenic spacer region (ITS). The three major methods produced highly concordant results, corroborating the view that each M. kansasii subtype does represent a distinct species. This work not only consolidates the position of five of the currently erected species, but also provides a description of the sixth one, i.e., Mycobacterium ostraviense sp. nov. to replace the former subtype IV. By showing a close genetic relatedness, a monophyletic origin, and overlapping phenotypes, our findings support the recognition of the M. kansasii complex (MKC), accommodating all M. kansasii-derived species and Mycobacterium gastri. None of the most commonly used taxonomic markers was shown to accurately distinguish all the MKC species. Likewise, no species-specific phenotypic characteristics were found allowing for species differentiation within the complex, except the non-photochromogenicity of M. gastri. To distinguish, most reliably, between the MKC species, and between M. kansasii and M. persicum in particular, whole-genome-based approaches should be applied. In the absence of clear differences in the distribution of the virulence-associated region of difference 1 genes among the M. kansasii-derived species, the pathogenic potential of each of these species can only be speculatively assessed based on their prevalence among the clinically relevant population. Large-scale molecular epidemiological studies Jagielski et al. Genomic Insights Into Mycobacterium kansasii Complex are needed to provide a better understanding of the clinical significance and pathobiology of the MKC species. The results of the in vitro drug susceptibility profiling emphasize the priority of rifampicin administration in the treatment of MKC-induced infections, while undermining the use of ethambutol, due to a high resistance to this drug.