Changes in Inflammatory Markers in Patients Treated for Buruli Ulcer and Their Ability to Predict Paradoxical Reactions.

Mycobacterium ulcerans causes Buruli ulcer, the third most frequent mycobacterial disease after tuberculosis and leprosy. Transient clinical deteriorations, known as paradoxical reactions (PRs), occur in some patients during or after antibiotic treatment. We investigated the clinical and biological features of PRs in a prospective cohort of 41 patients with Buruli ulcer from Benin. Neutrophil counts decreased from baseline to day 90, and interleukin 6 (IL-6), granulocyte colony-stimulating factor, and vascular endothelial growth factor were the cytokines displaying a significant monthly decrease relative to baseline. PRs occurred in 10 (24%) patients. The baseline biological and clinical characteristics of the patients presenting with PRs did not differ significantly from those of the other patients. However, the patients with PRs had significantly higher IL-6 and tumor necrosis factor alpha (TNF-α) concentrations on days 30, 60, and 90 after the start of antibiotic treatment. The absence of a decrease in IL-6 and TNF-α levels during treatment should alert clinicians to the possibility of PR onset.

Epidemiology of Nontuberculous Mycobacteria in French Polynesia.

As few data are available in the Pacific countries and territories of the Oceania region regarding nontuberculous mycobacteria,we retrospectively identified 87 such isolates from French Polynesia from 2008 to 2013 by hybridization using DNA-strip, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and partial rpoB gene sequencing. Partial rpoB gene sequencing classified 42/87 (48.3%) isolates in the Mycobacterium fortuitum complex, 28 (32.2%) in the Mycobacterium abscessus complex, 8 (9.2%) in the Mycobacterium mucogenicum complex, and 5 (5.7%) in the Mycobacterium avium complex. Two isolates were identified as Mycobacterium acapulcensis and Mycobacterium cosmeticum by partial 16S rRNA gene sequencing. One isolate, unidentified by MALDI-TOF MS and yielding less than 92% and 96% sequence similarity with rpoB and hsp65 reference sequences, respectively, was regarded as a potentially new species. Samples from three patients exhibiting>2 Mycobacterium porcinum isolates and from one patient with emphysema and a lung abscess exhibiting 2 Mycobacterium senegalense isolates fulfilled the American Thoracic Society microbiological criteria for nontuberculous mycobacterial lung infection.Remote geographic areas, such as French Polynesia, are potential sources for the discovery of new mycobacterial species.

Prevalence of Mycobacterium lentiflavum in cystic fibrosis patients, France.

BACKGROUND: Mycobacterium lentiflavum is rarely isolated in respiratory tract samples from cystic fibrosis patients. We herein describe an unusually high prevalence of M. lentiflavum in such patients. METHODS: M. lentiflavum, isolated from the respiratory tract of cystic fibrosis patients, was identified using both rpoB partial sequencing and detected directly in the sputum by using real-time PCR targeting the smpB gene. RESULTS: M. lentiflavum emerged as the third most prevalent nontuberculous mycobacterial species isolated in cystic fibrosis patients in Marseille, France. Six such patients were all male, and two of them may have fulfilled the American Thoracic Society clinical and microbiological criteria for M. lentiflavum potential lung infection. CONCLUSIONS: M. lentiflavum was the third most common mycobacteria isolated in cystic fibrosis patients, particularly in six male patients. M. lentiflavum outbreaks are emerging particularly in cystic fibrosis patients.

Bronchial colonisation in patients with lung cancer: a prospective study.

Bronchial colonisation is frequently reported in patients with lung cancer, and has a potential impact on therapeutic management and prognosis. We aimed to prospectively define the prevalence and nature of bronchial colonisation in patients at the time of diagnosing lung cancer. 210 consecutive patients with lung cancer underwent a flexible bronchoscopy for lung cancer. The type and frequency of bacterial, mycobacterial and fungal colonisation were analysed and correlated with the patients' and tumours' characteristics. Potential pathogens were found in 48.1% of samples: mainly the Gram-negative bacilli Escherichia coli (8.1%), Haemophilus influenzae (4.3%) and Enterobacter spp. (2.4%); Gram-positive cocci, Staphylococcus spp. (12.9%) and Streptococcus pneumoniae (3.3%); atypical mycobacteria (2.9%); Candida albicans (42.9%); and Aspergillus fumigatus (6.2%). Aged patients (p=0.02) with chronic obstructive pulmonary disease (p=0.008) were significantly more frequently colonised; however, tumour stage, atelectasis, bronchial stenosis and abnormalities of chest radiography were not associated with a higher rate of colonisation. Squamous cell carcinoma tended to be more frequently colonised than other histological subtypes. Airway colonisation was reported in almost half of patients presenting with lung cancer, mainly in fragile patients, and was significantly associated with worse survival (p=0.005). Analysing colonisation status of patients at the time of diagnosis may help improve the management of lung cancer.

"Mycobacterium mephinesia", a Mycobacterium terrae complex species of clinical interest isolated in French Polynesia.

A 59-year-old tobacco smoker male with chronic bronchitis living in Taravao, French Polynesia, Pacific, presented with a two-year growing nodule in the middle lobe of the right lung. A guided bronchoalveolar lavage inoculated onto Löwenstein-Jensen medium yielded colonies of a rapidly-growing non-chromogenic mycobacterium designed as isolate P7213. The isolate could not be identified using routine matrix-assisted laser desorption ionization-time of flight-mass spectrometry and phenotypic and probe-hybridization techniques and yielded 100% and 97% sequence similarity with the respective 16S rRNA and rpoB gene sequences of Mycobacterium virginiense in the Mycobacterium terrae complex. Electron microscopy showed a 1.15 µm long and 0.38 µm large bacillus which was in vitro susceptible to rifampicin, rifabutin, ethambutol, isoniazid, doxycycline and kanamycin. Its 4,511,948-bp draft genome exhibited a 67.6% G + C content with 4,153 coding-protein genes and 87 predicted RNA genes. Genome sequence-derived DNA-DNA hybridization, OrthoANI and pangenome analysis confirmed isolate P7213 was representative of a new species in the M. terrae complex. We named this species "Mycobacterium mephinesia".

Diversity of Mycobacterium tuberculosis lineages in French Polynesia.

BACKGROUND/PURPOSE: French Polynesia is an overseas territory located in the South Pacific. The incidence of tuberculosis in French Polynesia has been stable since 2000 with an average of 20 cases/y/100,000 inhabitants. Molecular epidemiology of Mycobacterium tuberculosis in French Polynesia is unknown because M. tuberculosis isolates have not been routinely genotyped. METHODS: From 2009 to 2012, 34 isolates collected from 32 French Polynesian patients were identified as M. tuberculosis by probe hybridization. These isolates were genotyped using spoligotyping and 24-loci mycobacterial interspersed repetitive units (MIRUs)-variable number of tandem repeat (VNTR). Spoligotype patterns obtained using commercial kits were compared with the online international database SITVIT. MIRU-VNTR genotyping was performed using an in-house protocol based on capillary electrophoresis sizing for 24-loci MIRU-VNTR genotyping. RESULTS: The results of the spoligotyping method revealed that 25 isolates grouped into six previously described spoligotypes [H1, H3, U likely (S), T1, Manu, and Beijing] and nine isolates grouped into six new spoligotypes. Comparison with the international database MIRU-VNTRplus distributed 30 isolates into five lineages (Haarlem, Latin American Mediterranean, S, X, and Beijing) and four as unassigned isolates. CONCLUSION: Genotyping identified four phylogenetic lineages belonging to the modern Euro-American subgroup, one Beijing genotype responsible for worldwide pandemics, including remote islands in the South Pacific, and one Manu genotype of the ancestral lineage of M. tuberculosis.

Cystic fibrosis respiratory tract salt concentration: An Exploratory Cohort Study.

In cystic fibrosis patients, electrolytic and osmolality imbalance secondary to cystic fibrosis transmembrane conductance regulator mutations may impact on mucoid secretion accumulation and secondary colonization by opportunistic pathogens such as nontuberculous mycobacteria.We performed a noninvasive exploratory prospective controlled clinical study comparing sputum salinity and acid-base characteristics of cystic fibrosis and noncystic fibrosis control patients. A total of 57 patients and 62 controls were included.Sputum salt concentrations were 10.5 g/L (95% CI: 7.7-13.3) in patients and 7.4 g/L (95% CI: 5.9-8.9) in aged-matched controls, a difference that was found to be statistically significant (P < .05). No difference in pH was observed between patients and controls.These differences in respiratory secretions salt concentrations could influence host-pathogen interactions in the context of cystic fibrosis respiratory infections. We propose to include respiratory secretion salt measurement as a routine analysis on cystic fibrosis patients' sputum submitted for bacterial culture.

Inverse correlation between salt tolerance and host-adaptation in mycobacteria.

BACKGROUND: The genus Mycobacterium includes host-adapted organisms regarded as obligate and opportunistic pathogens and environmental organisms. Factors contributing to this wide range of adaptations are poorly known. RESULTS: We studied the salt tolerance of 46 Mycobacterium species of medical interest. Representative strains of the Mycobacterium tuberculosis complex, Mycobacterium avium complex, Mycobacterium chelonae-abscessus complex, Mycobacterium ulcerans, Mycobacterium marinum, Mycobacterium lentiflavum, Mycobacterium fortuitum and Mycobacterium conceptionense were inoculated on Middlebrook 7H10 medium supplemented with 0-10% sodium chloride. Colonies were counted after 2-4 week incubation at the appropriate 30-37 °C temperature depending on the tested strain. Further comparative genomics was done on 15 Mycobacterium strains representing the spectrum of salt-tolerance of mycobacteria. Based on the results the different species were grouped according to their salt tolerance into a "salt-sensitive" group (growth up to ≤3% salt) containing the M. tuberculosis complex, Mycobacterium chelonae, Mycobacterium lentiflavum, Mycobacterium ulcerans and Mycobacterium marinum; a "salt-intermediate" group (growth between 4 and 6% salt) comprising Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium chimaera and a "salt-resistant" group (growth up to >6%) comprising Mycobacterium homonissuis, Mycobacterium bolettii, Mycobacterium fortuitum and Mycobacterium conceptionense. Genomic analysis revealed that 290 genes were unique to species belonging to the salt-sensitive group; and that 15% were annotated as being functionally associated with the ESX secretion systems Pro-Glu and Pro-Pro-Glu family proteins. CONCLUSIONS: In this work we found an inverse correlation between salt tolerance and host adaptation. We thus propose that salinity is one of the multiple factors determining the ecological niches of mycobacteria.

Draft Genome Sequence of Mycobacterium bohemicum Strain DSM 44277T.

The Mycobacterium bohemicum strain is a nontuberculosis species mainly responsible for pediatric cervical lymphadenitis. The draft genome of M. bohemicum DSM 44277(T) comprises 5,097,190 bp exhibiting a 68.64% G+C content, 4,840 protein-coding genes, and 75 predicted RNA genes.

Draft Genome Sequence of Mycobacterium lentiflavum CSUR P1491.

We announce the draft genome sequence of Mycobacterium lentiflavum strain CSUR P1491, a nontuberculous mycobacterium responsible for opportunistic potentially life-threatening infections in immunocompromised patients. The genome described here comprises a 6,818,507-bp chromosome exhibiting a 65.75% G+C content, 6,354 protein-coding genes, and 75 RNA genes.

Draft Genome Sequence of Mycobacterium europaeum Strain CSUR P1344.

We report the draft genome sequence of Mycobacterium europaeum strain CSUR P1344, a slowly growing mycobacterium of the Mycobacterium simiae complex and opportunistic respiratory tract colonizer and pathogen. This genome of 6,152,523 bp exhibits a 68.18% G+C content, encoding 5,814 predicted proteins and 74 RNAs.

Draft Genome Sequence of Mycobacterium tuberculosis Strain MT11, Which Represents a New Lineage.

We sequenced the genome of Mycobacterium tuberculosis strain MT11, which exhibits a specific 16S rRNA gene mutation found in 6% of French Polynesian M. tuberculosis isolates. It comprises a 4,110,293-bp chromosome with 65.15% G+C content, and it encodes 3,949 proteins and contains 85 predicted RNA genes. The TbD1 region is absent in strain MT11 as in modern M. tuberculosis strains.

Draft Genome Sequence of Mycobacterium tuberculosis Strain MT43, a Representative of the Manu2 Genotype.

We announce the draft genome sequence of Mycobacterium tuberculosis strain MT43, isolated from a pulmonary form of tuberculosis in French Polynesia. Analyzing its 4,145,007-bp, 65.17% G+C chromosome confirmed a fully antibiotic-susceptible Manu2 spoligotype.

Respiratory tract isolation of Mycobacterium europaeum following influenza infection in an immunocompromised patient: a case report.

INTRODUCTION: Mycobacterium europaeum, a slow-growing nontuberculous mycobacteria belonging to the Mycobacterium simiae complex, was described after the seminal characterization of five isolates collected from three sputum specimens and a jaw gland biopsy in Italy, Greece and Sweden. Five respiratory tract isolates were further reported in Iran. Here, we report the first isolation of M. europaeum in France, in the respiratory tract of a patient co-infected with human immunodeficiency virus and hepatitis C virus. CASE PRESENTATION: A 49-year-old Caucasian woman with a 26-year history of human immunodeficiency virus-hepatitis C virus co-infection was admitted for significant influenza-like syndrome in a context of repetitive exacerbations of chronic obstructive pulmonary disease. Significant biological parameters included lymphocytes of 1.6G/L including 237/mm3 T4 lymphocytes, a human immunodeficiency virus viral load of 1.6 log and a hepatitis C virus viral load of 6 log. Reverse-transcriptase polymerase chain reaction of her nasopharyngeal aspiration confirmed influenza A H1N1. Three sputum specimens lacked acid-fast bacilli but one grew mycobacteria identified by using matrix-assisted laser desorption ionization/time-of-flight mass spectrometry as M. europaeum with a 1.56 log score. A 1,482-bp 16S ribosomal ribonucleic acid gene sequence yielded 99% similarity with both Mycobacterium parascrofulaceum ATCC BAA-614 and M. europaeum DSM 45397T and partial rpoB polymerase chain reaction-sequencing yielded a 725-bp sequence exhibiting 100% similarity with M. europaeum strain DSM 45397T. CONCLUSIONS: We report the first isolation of M. europaeum in France, in the respiratory tract of a patient co-infected with human immunodeficiency virus and hepatitis C virus. M. europaeum warrants further attention in immunosuppressed patients with influenza, using matrix-assisted laser desorption ionization/time-of-flight mass spectrometry and rpoB partial sequencing as tools for its accurate identification.

Draft Genome Sequence of Mycobacterium neoaurum Strain DSM 44074T.

We report the draft genome sequence of Mycobacterium neoaurum strain DSM 44074(T), a nontuberculosis species responsible for opportunistic infections in immunocompromised patients. The strain described here is composed of 5,536,033 bp, with a G+C content of 66.24%, and carries 5,274 protein-coding genes and 72 RNA genes.