The Chemical Composition and Anti-mycobacterial Activities of Trachyspermum copticum and Pelargonium graveolens Essential Oils.

BACKGROUND: Microbial resistance to antibiotics and their adverse effects related to these antibiotics are a matter of global public health in the 21th century. The emergence of drug-resistant strains, has gained the interest of the scientists to discover new antimicrobial agents from the essential oil of medicinal plants. METHODS: Anti-mycobacterial effects of Trachyspermum copticum and Pelargonium graveolens essential oils were determined against multi-drug resistant clinical strains of Mycobacterium tuberculosis, Mycobacterium kansasii, Mycobacterium fortuitum and standard strain of Mycobacterium tuberculosis H37Rv by a Broth micro-dilution method. Pelargonium graveolens plant named Narmada was discovered by Kulkarni R.N et al. (Patent ID, USPP12425P2) and a formulation comprising thymol obtained from Trachyspermum is useful in the treatment of drug-resistant bacterial infections (Patent ID, US6824795B2). The chemical composition of hydro-distilled essential oils was determined by GC and GC-MS. RESULTS: Minimum Inhibitory Concentration (MIC) values for T. copticum essential oil against tested isolates were ranged from 19.5 µg/mL to 78 µg/mL. The least minimum inhibitory concentration of P. graveolens extract against M. Kansasii and MDR-TB was 78 µg/ml. CONCLUSION: The results of the present research introduced T. copticum and P. graveolens essential oils as a remarkable natural anti-mycobacterial agent, but more pharmacological studies are required to evaluate their efficacy in animal models.

Abdominal violaceous skin lesions of a 47-year-old woman following a geometric pattern / Lesiones cutáneas violáceas en el abdomen de una mujer de 47 años siguiendo una distribución geométrica

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Métodos de determinación de sensibilidad a los antimicrobianos en micobacterias / Methods for determining the antimicrobial susceptibility of mycobacteria

Las micobacterias son un amplio grupo de microorganismos en el que múltiples especies son causa de una importante morbimortalidad, como la tuberculosis y la lepra. La aparición y diseminación de cepas del complejo Mycobacterium tuberculosis resistentes a diversos fármacos constituye en la actualidad uno de los problemas sanitarios de mayor gravedad a nivel mundial. Por otro lado, las micobacterias diferentes de M. tuberculosis y Mycobacterium leprae, denominadas micobacterias no tuberculosas (MNT), son aislamientos cada vez más frecuentes, requiriendo en muchos casos un tratamiento que precisa una orientación sobre la sensibilidad de estos microorganismos a los antimicrobianos. En el presente artículo se revisan los métodos para determinar la sensibilidad in vitro a los antimicobacterianos de los aislamientos del complejo M. tuberculosis y las MNT más relevantes. Además, también se realiza un análisis de las técnicas moleculares de detección rápida de la resistencia a partir de las muestras clínicas (AU)

Mycobacteria are a large group of microorganisms, multiple species of which are major causes of morbidity and mortality, such as tuberculosis and leprosy. At present, the emergence and spread of multidrug-resistant strains of Mycobacterium tuberculosis complex are one of the most serious health problems worldwide. Furthermore, in contrast to M. tuberculosis and Mycobacterium leprae, non-tuberculous mycobacteria (NTM) are more frequently isolated and, in many cases, treatment is based on drug susceptibility testing. This article is a review of the different methods to determine the in vitro drug susceptibility of M. tuberculosis complex and the most relevant NTM isolates. The molecular techniques currently used for rapid detection of resistance of clinical specimens are also analysed (AU)

Mycobacterium stephanolepidis sp. nov., a rapidly growing species related to Mycobacterium chelonae, isolated from marine teleost fish, Stephanolepis cirrhifer.

A previously undescribed rapidly growing, non-pigmented mycobacterium was identified based on biochemical and nucleic acid analyses, as well as growth characteristics. Seven isolates were cultured from samples collected from five thread-sail filefish (Stephanolepis cirrhifer) and two farmed black scraper (Thamnaconus modestus). Bacterial growth occurred at 15-35 °C on Middlebrook 7H11 agar. The bacteria were positive for catalase activity at 68 °C and urease activity, intermediate for iron uptake, and negative for Tween 80 hydrolysis, nitrate reduction, semi-quantitative catalase activity and arylsulfatase activity at day 3. No growth was observed on Middlebrook 7H11 agar supplemented with picric acid, and very little growth was observed in the presence of 5 % NaCl. α- and α'-mycolates were identified in the cell walls, and a unique profile of the fatty acid methyl esters and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiles of the protein and cell-wall lipids were acquired. Sequence analysis revealed that the seven isolates shared identical sequences for the 16S rRNA, rpoB, hsp65, recA and sodA genes. Phylogenetic analysis of the five gene sequences confirmed that the isolates were unique, but closely related to Mycobacterium chelonae. Antibiotic susceptibility testing revealed the minimum inhibitory concentration (MIC) of clarithromycin against this novel species was <0.25 µg ml-1, which was lower than that for Mycobacterium salmoniphilum. The hsp65 PCR restriction enzyme analysis pattern differed from those of M. chelonae and M. salmoniphilum. Based on these findings, the name Mycobacterium stephanolepidis sp. nov. is proposed for this novel species, with the type strain being NJB0901T (=JCM 31611T=KCTC 39843T).

Drug Susceptibility Testing of 31 Antimicrobial Agents on Rapidly Growing Mycobacteria Isolates from China.

OBJECTIVES: Several species of rapidly growing mycobacteria (RGM) are now recognized as human pathogens. However, limited data on effective drug treatments against these organisms exists. Here, we describe the species distribution and drug susceptibility profiles of RGM clinical isolates collected from four southern Chinese provinces from January 2005 to December 2012. METHODS: Clinical isolates (73) were subjected to in vitro testing with 31 antimicrobial agents using the cation-adjusted Mueller-Hinton broth microdilution method. The isolates included 55 M. abscessus, 11 M. fortuitum, 3 M. chelonae, 2 M. neoaurum, and 2 M. septicum isolates. RESULTS: M. abscessus (75.34%) and M. fortuitum (15.07%), the most common species, exhibited greater antibiotic resistance than the other three species. The isolates had low resistance to amikacin, linezolid, and tigecycline, and high resistance to first-line antituberculous agents, amoxicillin-clavulanic acid, rifapentine, dapsone, thioacetazone, and pasiniazid. M. abscessus and M. fortuitum were highly resistant to ofloxacin and rifabutin, respectively. The isolates showed moderate resistance to the other antimicrobial agents. CONCLUSIONS: Our results suggest that tigecycline, linezolid, clofazimine, and cefmetazole are appropriate choices for M. abscessus infections. Capreomycin, sulfamethoxazole, tigecycline, clofazimine, and cefmetazole are potentially good choices for M. fortuitum infections. Our drug susceptibility data should be useful to clinicians.

Infections Caused by Rapidly Growing Mycobacteria spp in Children and Adolescents With Cancer.

BACKGROUND: Rapidly growing mycobacteria (RGM) infections in pediatric oncology patients have not been completely characterized. METHODS: We reviewed medical records of oncology patients at St. Jude Children's Research Hospital (St. Jude) from 1990 to 2010 with RGM infections and summarized the results of previously published cases. RESULTS: Twenty-five St. Jude patients had 27 episodes of infection. Approximately half of the cases occurred in patients with hematological malignancies and in males; infections were more common in white patients. Most patients were not neutropenic or lymphopenic. The most common causative species were Mycobacterium chelonae, Mycobacterium abscessus, and Mycobacterium fortuitum. Most isolates were susceptible to amikacin and clarithromycin; all were susceptible to at least 1 of these. Treatment regimens varied considerably, particularly with respect to the duration of antimicrobial chemotherapy. Two St. Jude patients died; both had pulmonary infections. The literature search identified an additional 58 cases of infection. Localized catheter-associated infections were more common than bloodstream infections in the current series than in previous reports, and outbreaks were not recognized. Otherwise, the demographic and clinical characteristics of patients were similar. CONCLUSIONS: Localized catheter-associated infections were most common in this largest reported single center experience reported to date. Pulmonary infection is uncommon in children but, as in adults, has a high mortality rate. Relatively short-term antimicrobial treatment and surgical debridement of infected tissue, if present, may be as effective for catheter-associated infections as prolonged antimicrobial use and may reduce adverse drug effects in these patients, who are vulnerable to drug-drug interactions and toxicity.

Two-Year Monitoring of Water Samples from Dam of Iskar and the Black Sea, Bulgaria, by Molecular Analysis: Focus on Mycobacterium spp.

The coast of the Bulgarian Black Sea is a popular summer holiday destination. The Dam of Iskar is the largest artificial dam in Bulgaria, with a capacity of 675 million m3. It is the main source of tap water for the capital Sofia and for irrigating the surrounding valley. There is a close relationship between the quality of aquatic ecosystems and human health as many infections are waterborne. Rapid molecular methods for the analysis of highly pathogenic bacteria have been developed for monitoring quality. Mycobacterial species can be isolated from waste, surface, recreational, ground and tap waters and human pathogenicity of nontuberculose mycobacteria (NTM) is well recognized. The objective of our study was to perform molecular analysis for key-pathogens, with a focus on mycobacteria, in water samples collected from the Black Sea and the Dam of Iskar. In a two year period, 38 water samples were collected-24 from the Dam of Iskar and 14 from the Black Sea coastal zone. Fifty liter water samples were concentrated by ultrafiltration. Molecular analysis for 15 pathogens, including all species of genus Mycobacterium was performed. Our results showed presence of Vibrio spp. in the Black Sea. Rotavirus A was also identified in four samples from the Dam of Iskar. Toxigenic Escherichia coli was present in both locations, based on markers for stx1 and stx2 genes. No detectable amounts of Cryptosporidium were detected in either location using immunomagnetic separation and fluorescence microscopy. Furthermore, mass spectrometry analyses did not detect key cyanobacterial toxins. On the basis of the results obtained we can conclude that for the period 2012-2014 no Mycobacterium species were present in the water samples. During the study period no cases of waterborne infections were reported.

Antimycobacterial activity of Citrullus colocynthis (L.) Schrad. against drug sensitive and drug resistant Mycobacterium tuberculosis and MOTT clinical isolates.

ETHNOPHARMACOLOGICAL RELEVANCE: Citrullus colocynthis (Cucurbitaceae), a folk herbal medicine and traditionally used natural remedy for tuberculosis in India has been studied to validate its antitubercular activity against drug sensitive and drug resistant (including multidrug resistant) Mycobacterium tuberculosis and Mycobacterium other than tuberculosis (MOTT) bacilli. MATERIALS AND METHODS: Inhibitory and bactericidal activities of crude extracts, fractions and compounds of Citrullus colocynthis plant, consisting of aerial parts and ripe deseeded fruits were performed against the drug sensitive standard strain of Mycobacterium tuberculosis H37Rv (ATCC 27294), 16 drug resistant strains of Mycobacterium tuberculosis and two MOTT strains, using radiometric BACTEC 460TB system. RESULTS: Methanolic extract of ripe deseeded fruit of Citrullus colocynthis has shown good activity (MIC ≤ 62.5 µg/ml), whereas among the bioactive fractions, FC IX demonstrated the best activity (MIC 31.2 µg/ml) against Mycobacterium tuberculosis H37Rv. Bioactive FC III, IX and X also inhibited 16 clinical isolates of Mycobacterium tuberculosis consisting of seven non-multidrug resistants, eight multidrug resistants, one extensively drug resistant and two of MOTTs with MICs in the range of 50-125, 31.2-125 and 62.5-125 µg/ml, respectively. Ursolic acid and cucurbitacin E 2-0-ß-d-glucopyranoside were identified as the main biomarkers active against Mycobacterium tuberculosis H37Rv (MICs 50 and 25 µg/ml respectively), as well as against the 18 clinical isolates. FC III and FC IX showed better inhibition of drug resistant and MOTT clinical isolates. Minimal bactericidal concentrations of extracts, fractions and compound C-2 were ≥ two-fold MIC values. CONCLUSIONS: The study provides a scientific rationale for the traditional use of Citrullus colocynthis fruit in the treatment of tuberculosis. In addition, the study elucidates a broad spectrum antimycobacterial action of Citrullus colocynthis fruit, which can contribute to the development of improved preparation of an antitubercular natural drug for the treatment of drug resistant tuberculosis and MOTT infection as well.

[Optimization of complex treatment in ocular toxoplasmosis and concurrent infections].

137 patients (177 eyes) with verified toxoplasmic uveitis, retinitis, chorioretenitis were observed. Among them 65 patients had concurrent infections: tuberculosis, herpes simplex and chlamydia. Routine ophthalmologic, clinical and laboratory examination was performed. The results of intensive treatment in acute and chronic phases are presented, the staged drug pathogenic treatment including methods of specific therapy, based on differential approach to anti-inflammatory agents use, was provided. Early diagnosis and appropriate management including combined treatment of concurrent infections improves treatment efficacy and allows to achieve excellent results.

Drug susceptibility testing and pharmacokinetics question current treatment regimens in Mycobacterium simiae complex disease.

The Mycobacterium simiae complex bacteria can cause opportunistic infections in humans. In the case of definite disease, there are no evidence-based treatment regimens and outcomes are very disappointing. To increase the evidence base underpinning treatment regimens for M. simiae complex disease, drug susceptibility patterns and rifampicin/ethambutol synergy were assessed retrospectively in 69 clinical M. simiae complex isolates from 60 patients (22 patients with M. simiae, 24 with Mycobacterium lentiflavum, 8 with Mycobacterium triplex, 5 with Mycobacterium parascrofulaceum and 1 with Mycobacterium stomatepiae) submitted to the mycobacteriology laboratory at National Jewish Health (Denver, CO). Quantitative drug susceptibility testing (DST) was performed using the radiometric BacTec 460 macrodilution method. Results were related to pharmacokinetic (PK) measurements, where available. All M. simiae complex species proved susceptible to clarithromycin and, to a lesser extent, rifabutin, clofazimine, streptomycin and moxifloxacin. Synergy or additive action between rifampicin and ethambutol was observed for all species except M. simiae. Mycobacterium simiae is poorly susceptible in vitro to rifampicin and ethambutol alone as well as in combination; PK measurements support the limited efficacy of these drugs against M. simiae. The triple-drug regimen of a rifamycin, ethambutol and a macrolide may be advised to treat disease caused by M. lentiflavum, M. triplex, M. parascrofulaceum and M. stomatepiae; for M. simiae, this regimen appears less active. These findings may partly explain the limited treatment results in M. simiae disease. A treatment regimen including a macrolide, moxifloxacin and one or two additional drugs based on DST results may be advisable; clofazimine and amikacin or streptomycin are potential candidates.

Aerobic biotransformation of alkyl branched aromatic alkanoic naphthenic acids via two different pathways by a new isolate of Mycobacterium.

Naphthenic acids (NAs) are complex mixtures of carboxylic acids found in weathered crude oils and oil sands, and are toxic, corrosive and persistent. However, little is known about the microorganisms and mechanisms involved in NA degradation. We isolated a sediment bacterium (designated strain IS2.3), with 100% 16S rRNA gene sequence identity to Mycobacterium aurum, which degraded synthetic NAs (4'-n-butylphenyl)-4-butanoic acid (n-BPBA) and (4'-t-butylphenyl)-4-butanoic acid (t-BPBA). n-BPBA was readily oxidized with almost complete degradation (96.8% ± 0.3) compared with t-BPBA (77.8% ± 3.7 degraded) by day 49. Cell counts increased fourfold by day 14 but decreased after day 14 for both n- and t-BPBA. At day 14, (4'-butylphenyl)ethanoic acid (BPEA) metabolites were detected. Additional metabolites produced during t-BPBA degradation were identified by mass spectrometry of derivatives as (4'-carboxy-t-butylphenyl)-4-butanoic acid and (4'-carboxy-t-butylphenyl)ethanoic acid; suggesting that strain IS2.3 used omega oxidation of t-BPEA to oxidize the tert-butyl side-chain to produce (4'-carboxy-t-butylphenyl)ethanoic acid, as the primary route for biodegradation. However, strain IS2.3 also produced this metabolite through initial omega oxidation of the tert-butyl side-chain of t-BPBA, followed by beta-oxidation of the alkanoic acid side-chain. In conclusion, an isolate belonging to the genus Mycobacterium degraded highly branched aromatic NAs via two different pathways.

Tuberculosis in Australia: bacteriologically confirmed cases and drug resistance, 2008 and 2009. A report of the Australian Mycobacterium Reference Laboratory Network.

There were 886 and 1,062 bacteriologically-confirmed cases of tuberculosis (TB) in 2008 and 2009, representing an annual rate of 4.1 and 4.9 cases per 100,000 population respectively. Over the 2 years, a total of 23 children aged under 10 years (male n = 13, female n = 10) had bacteriologically confirmed tuberculosis, including 3 children with TB meningitis. Results of in vitro drug susceptibility testing were available for 885 of 886 and 1,060 of 1,062 isolates for isoniazid (INH), rifampicin (RIF), ethambutol (EMB), and pyrazinamide (PYZ) in 2008 and 2009 respectively. In 2008, a total of 94 (10.7%) isolates of Mycobacterium tuberculosis complex were resistant to at least one of the anti-tuberculosis agents. Any resistance to INH was noted for 76 (8.7%), 23 (2.6%) for RIF, 10 (1.1%) for EMB and 9 (1.0%) for PYZ. Resistance to at least INH and RIF (defined as multidrug-resistant TB (MDR-TB) was detected in 21 (2.4%) isolates. None of the 21 MDR-TB isolates had resistance to either ofloxacin or the injectable agents. In 2009, a total of 168 (15.9%) were resistant to at least one of the anti-TB agents. Any resistance to INH was noted for 150 (14.2%) isolates, 37 (3.5%) for RIF, 5 (0.5%) for EMB and 13 (1.2%) for PYZ. A total of 31 (2.9%) isolates were MDR-TB. In 2009, there were 2 cases of quinolone resistance in MDR-TB from persons born overseas. Mono-resistance to INH was the most commonly detected resistance with 33 and 80 isolates in 2008 and 2009, respectively. Mono-resistance to RIF was infrequently encountered with 2 and 5 isolates in 2008 and 2009 respectively. There were six and 11 MDR-TB patients from the Papua New Guinea (PNG) - Torres Strait Islands (TSI) cross-border region in 2008 and 2009 respectively. The PNG-TSI zone now contributes a substantial proportion of MDR-TB cases to the database. In addition, there were 24 isolates of Mycobacterium bovis bacille Calmette Guérin (BCG), 15 were cultured from males (4 aged < or = 5 years) and from 9 females (5 aged < or = 5 years). The predominant site of isolation was from vaccination abscess. Eight males (range: 57-87 years) had M. bovis BCG isolated from urine or blood culture.

An outbreak of cutaneous infection due to Mycobacterium abscessus associated to mesotherapy

Introduction: In February 2009 an outbreak of subcutaneous abscesses due to Mycobacterium abscessus was detected in Spain which affected healthy women who had undergone mesotherapy procedures in an aesthetic clinic. Methods Epidemiological research, health inspection and microbiological studies were conducted. The patients were given antibiotic treatment (according to susceptibility testing) with clarithromycin, and in some cases, combined with amikacin. Results Seventeen out of 77 patients treated in the clinic were affected. The products used for the injections were homeopathic drugs in multi-dose vials. The environmental samples were negative. The sterile injection equipment and the clinical procedures were evaluated as correct. The storage conditions for the drugs were also correct, and all the samples tested negative for Mycobacteria. However Paenibacillus provencensis was isolated from samples of unused multi-dose vials and the withdrawal of the product from distribution was ordered. Deficiencies were detected in the sterile products process of at the homeopathic drug factory, so the production line was suspended. Conclusions The results of environmental investigation suggest the most likely cause of the outbreak could have been the contamination of the products in the factory, although there was no laboratory confirmation. The widespread use of homeopathic products in invasive procedures requires extreme control during the manufacturing, handling and packaging process. It is important to consider mesotherapy and parenteral use of homeopathic medicines as potential sources of infection and therefore the same precautions in the procedures and quality assurance of products should be applied as with any other drug or medical activity (AU)

Introducción: En febrero de 2009 se detectó en Baleares un brote de abscesos subcutáneos causados por Mycobacterium abscessus que afectaba mujeres jóvenes y sanas que se habían sometido a procedimientos de mesoterapia en una clínica de estética. Métodos: Se realizaron investigación epidemiológica, inspección sanitaria y estudios microbiológicos clínicos y ambientales. Los pacientes iniciaron tratamiento antibiótico (según antibiograma) con claritromicinay, en los casos más graves, amikacina. Resultados: Aparecieron lesiones en 17 de las 77 personas sometidas a la mesoterapia en el período de riesgo. Los productos inyectados eran fármacos homeopáticos en multivial. Las muestras ambientales fueron negativas. No se evidenciaron deficiencias en los equipos y procedimientos. Los medicamentos estaban correctamente almacenados y todas las muestras fueron negativas para Mycobacteria, aunque se identificó Paenibacillus provencensis de multiviales precintados y el producto fue retirado de la distribución. Se detectaron deficiencias en la producción de estériles en la fábrica, por lo que la línea de producción fue suspendida y el producto retirado. Conclusiones: Los resultados de la investigación ambiental sugieren que la causa más probable del brote habría sido la contaminación del producto en origen, aunque no fue confirmada por laboratorio. La difusión del uso de productos homeopáticos en procedimientos invasivos requiere un control riguroso durante la fabricación, manipulación y envasado. Es importante considerar la mesoterapia y el uso parenteral de productos homeopáticos como fuentes potenciales de infección y por lo tanto extremar las precauciones y la garantía de calidad de los productos y los procedimientos de la misma manera que con cualquier otro producto farmacológico o actividad médica (AU)

Clarithromycin and amikacin vs. clarithromycin and moxifloxacin for the treatment of post-acupuncture cutaneous infections due to Mycobacterium abscessus: a prospective observational study.

An outbreak of post-acupuncture cutaneous infections due to Mycobacterium abscessus occurred in Ansan, Korea, from November 2007 through to May 2008. During this time a prospective, observational, non-randomized study was conducted involving 52 patients that were diagnosed with cutaneous M. abscessus infection. We compared the clinical response between patients treated with clarithromycin plus amikacin regimen and those treated with clarithromycin plus moxifloxacin regimens with regard to time to resolution of the cutaneous lesions. Among the 52 study patients, 33 were treated with clarithromycin plus amikacin, and 19 were treated with clarithromycin plus moxifloxacin. The baseline characteristics for the treatment groups were not significantly different, except for initial surgical excision (n = 27 vs. 6, respectively, p = 0.001). The median time (weeks) to resolution of the lesions in the clarithromycin plus moxifloxacin-treated subjects was significantly shorter than that in the clarithromycin plus amikacin-treated subjects (17 ± 1.1 vs. 20 ± 0.9, respectively, p = 0.017). With adjustments for age, location of lesions, prior incision and drainage, and excision during medical therapy, clarithromycin plus moxifloxacin-treated subjects were more likely to have resolved lesions (hazard ratio, 0.387; 95% confidence interval, 0.165-0.907; p = 0.029). The frequency of drug-related adverse events in the two treatment groups was not significantly different (n = 18 vs. 14, respectively; p = 0.240). The most common adverse event was gastrointestinal discomfort. The results of our study showed that the combination regimen of clarithromycin and moxifloxacin resulted in a better clinical response than a regimen of clarithromycin plus amikacin when used for treatment of cutaneous M. abscessus infection.

Tres casos de paniculitis por Mycobacterium abscessus posmesoterapia / Three Cases of Panniculitis due to Mycobacterium Abscessus After Mesotherapy

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Mycobacterium microti infection associated with spindle cell pseudotumour and hypercalcaemia: a possible link with an infected alpaca.

A 44-year-old woman who had recently been on immunosuppressive therapy presented with malaise, cough, fever, weight loss, lymphadenopathy, severe hypercalcaemia and a paratracheal mass on imaging. The initial impression was of disseminated malignancy, and lymphoma was suspected. A mediastinal biopsy showed a mycobacterial spindle cell pseudotumour containing acid and alcohol fast bacilli (AAFB). Sputum microscopy demonstrated AAFBs, confirmed as Mycobacterium tuberculosis complex by PCR. Prolonged culture grew Mycobacterium microti, an organism often associated with disease in small rodents and llamas. M microti isolates from postmortem samples of an alpaca at a nearby farm were genetically indistinguishable. Although the patient had not visited the farm, concurrent illness in her adopted stray cat suggested a possible zoonotic connection. The patient responded to antituberculous therapy, and rehydration and pamidronate for hypercalcaemia. We believe the hypercalcaemia was caused by a similar mechanism to raised calcium levels sometimes seen in tuberculosis.

Mycobacterium phocaicum in therapy pool water.

We isolated Mycobacterium phocaicum, an emerging non-tuberculous species rarely identified in the respiratory tract and blood of patients, from the therapy pool water. Identification, confirmed by 16S rDNA and rpoB sequencing and phylogenetic analyses, disclosed a genotype 4. M. phocaicum should be added to the growing list of water-borne mycobacteria.

Diagnosis and treatment of infections caused by rapidly growing mycobacteria.

Rapidly growing mycobacteria (RGM) are ubiquitous environmental organisms capable of causing a wide variety of infections in humans. The prevalence of RGM infections appears to be increasing, although exact incidence rates are unknown. Although some risk factors for pulmonary RGM infection have been determined, the specific host factors predisposing to disease in the majority of cases are not clear. Significant advances in molecular methods of mycobacterial identification have led to isolation of more varieties, changes in taxonomy, and more rapid and accurate diagnosis of RGM from clinical isolates. Despite significant advances in the field, diagnosing and treating RGM pulmonary infections remain complicated. Current guidelines are based on the most commonly encountered NTM. Their applicability to less frequent RGM isolates has not been definitively established. Treatment often requires multiple antimicrobial agents for prolonged periods of time, with varying degrees of success and significant associated morbidity.

Molecular characterization of Mycobacterium massiliense and Mycobacterium bolletii in isolates collected from outbreaks of infections after laparoscopic surgeries and cosmetic procedures.

An outbreak of infections affecting 311 patients who had undergone different invasive procedures occurred in 2004 and 2005 in the city of Belém, in the northern region of Brazil. Sixty-seven isolates were studied; 58 were from patients who had undergone laparoscopic surgeries, 1 was from a patient with a postinjection abscess, and 8 were from patients who had undergone mesotherapy. All isolates were rapidly growing nonpigmented mycobacteria and presented a pattern by PCR-restriction enzyme analysis of the hsp65 gene with BstEII of bands of 235 and 210 bp and with HaeIII of bands of 200, 70, 60, and 50 bp, which is common to Mycobacterium abscessus type 2, Mycobacterium bolletii, and Mycobacterium massiliense. hsp65 and rpoB gene sequencing of a subset of 20 isolates was used to discriminate between these three species. hsp65 and rpoB sequences chosen at random from 11 of the 58 isolates from surgical patients and the postinjection abscess isolate presented the highest degrees of similarity with the corresponding sequences of M. massiliense. In the same way, the eight mesotherapy isolates were identified as M. bolletii. Molecular typing by pulsed-field gel electrophoresis (PFGE) grouped all 58 surgical isolates, while the mesotherapy isolates presented three different PFGE patterns and the postinjection abscess isolate showed a unique PFGE pattern. In conclusion, molecular techniques for identification and typing were essential for the discrimination of two concomitant outbreaks and one case, the postinjection abscess, not related to either outbreak, all of which were originally attributed to a single strain of M. abscessus.