Mycobacterium phlei, a previously unreported cause of pacemaker infection: thinking outside the box in cardiac device infections.

The increased use of cardiac rhythm management devices has led to an increase in cardiac device-related infections (CDI). Staphylococcus aureus and epidermidis account for the vast majority of CDI. CDI due to rapidly growing non-tuberculous mycobacteria is very rare, with only about ten cases having been reported. We report a case of pacemaker pocket infection with Mycobacterium phlei. There are only three published reports of human infection involving this typically non-pathogenic organism. To the best of our knowledge, this is the first report of CDI with Mycobacterium phlei.

Mycobacterial infections in human immuno-deficiency virus seropositive patients: role of non-tuberculous mycobacteria.

BACKGROUND: There is high prevalence of tuberculosis in patients with HIV infection; hence the role of non-tuberculous mycobacteria (NTM) in HIV patients has always been undermined. NTM may be responsible for clinical disease in a substantial number of immuno-compromised HIV sero-positive individuals even in a country endemic for Mycobacterium tuberculosis (M. tuberculosis). The study was designed to look for the contribution of NTM to morbidity in HIV seropositive patients. MATERIAL AND METHODS: In a prospective study of ninety-four HIV seropositive individuals presenting with pulmonary or extra-pulmonary symptoms suggestive of mycobacterial infection, appropriate samples were collected and processed. Detailed clinical history was utilized to differentiate colonization or contamination by NTM from true lung disease. RESULTS: Fourteen samples grew mycobacterial species, 8(57.2%) being NTM. The distribution of NTM was--3 M. avium complex, 2 M. fortuitum, 2 M. vaccae, 1 M. phlei. 6 isolates were M. tuberculosis. CONCLUSION: NTM may be responsible for a significant proportion of mycobacterial infections in HIV seropositive individuals. Despite the high endemicity of tuberculosis in developing countries like India, the presence of NTM should be ruled out; especially in immuno-compromised HIV seropositive individuals before instituting anti-tubercular therapy empirically. In addition, non-response of NTM to ATT may be wrongly attributed to multi-drug resistant tuberculosis.

[Comparison of three different mycobacterial DNA isolation methods]. / Mikobakteriyel DNA izolasyonunda üç farkli yönteminin karsilastirilmasi.

Rapid diagnosis of mycobacterial infections is of crucial importance. For this reason molecular methods have become valuable diagnostic tools. DNA isolation step prior to polymerase chain reaction (PCR) is one of the most important steps that affect the sensitivity of the diagnostic tests. In this study, three isolation methods which could affect the sensitivity of the PCR methods used for the diagnosis of mycobacterial infections were compared. For this reason the sensitivities of boiling method, MagNA Pure LC automated isolation system (Roche Applied Science, Germany) and Magtration 12GC (Precision System Science, Germany) automated isolation system were compared by using four different mycobacterial strains (Mycobacterium tuberculosis H37Ra standard strain, M. tuberculosis clinical isolate, M. phlei and M. smegmatis). DNAs were isolated from serial dilutions of these mycobacterial strains by using three isolation methods, and 441 base pair region of hsp65 gene was amplified by PCR. After the obtained products were run on agarose gel, it was observed that the lowest dilution rates at which bands could be seen were 10(-5) dilutions of H37Ra DNA isolated by MagNA Pure and Magtration systems. Isolation of H37Ra DNA by boiling method yielded a band in 10(-2) dilution sample. Isolation of DNA from Mycobacterium tuberculosis clinical isolate by MagNA Pure system, Magtration system and boiling method provided amplification in 10(-4), 10(-3) and 10(-1) dilution samples, respectively. The lowest dilution samples which yielded visible bands on agarose gel were 10(-5), 10(-4) and 10(-2) dilution samples for M. smegmatis strain and 10(-4), 10(-3) and 10(-2) dilution samples for M. phlei strain. The results of this study revealed that MagNA Pure system could detect smaller amounts of DNA in the sample while boiling method was not a reliable method to detect small amounts of DNA in clinical samples. Therefore it was concluded that the use of automated isolation systems could be a reliable alternative for routine PCR applications by providing both high sensitivity and standardization.

Thermophilic biodesulfurization of various heterocyclic sulfur compounds and crude straight-run light gas oil fraction by a newly isolated strain Mycobacterium phlei WU-0103.

Various heterocyclic sulfur compounds such as naphtho[2,1-b]thiophene (NTH) and benzo[b]thiophene (BTH) derivatives can be detected in diesel oil, in addition to dibenzothiophene (DBT) derivatives. Mycobacterium phlei WU-0103 was newly isolated as a bacterial strain capable of growing in a medium with NTH as the sulfur source at 50 degrees C. M. phlei WU-0103 could degrade various heterocyclic sulfur compounds, not only NTH and its derivatives but also DBT, BTH, and their derivatives at 45 degrees C. When M. phlei WU-0103 was cultivated with the heterocyclic sulfur compounds such as NTH, NTH 3,3-dioxide, DBT, BTH, and 4,6-dialkylDBTs as sulfur sources, monohydroxy compounds and sulfone compounds corresponding to starting heterocyclic sulfur compounds were detected by gas chromatography-mass spectrometry analysis, suggesting the sulfur-specific desulfurization pathways for heterocyclic sulfur compounds. Moreover, total sulfur content in 12-fold-diluted crude straight-run light gas oil fraction was reduced from 1000 to 475 ppm S, with 52% reduction, by the biodesulfurization treatment at 45 degrees C with growing cells of M. phlei WU-0103. Gas chromatography analysis with a flame photometric detector revealed that most of the resolvable peaks, such as those corresponding to alkylated derivatives of NTH, DBT, and BTH, disappeared after the biodesulfurization treatment. These results indicated that M. phlei WU-0103 may have a good potential as a biocatalyst for practical biodesulfurization of diesel oil.

Nontuberculous mycobacteria in soils of La Pampa province (Argentina).

The presence of nontuberculous mycobacteria (NTM) was investigated in forty soil samples belonging to the four physiographic regions (Eastern, Central, Southern and Western) that constitute La Pampa province. The presence of NTM in 67.5% of these soil samples was determined. The density of mycobacteria ranged 25-4,500 mycobacteria g-1 dry soil (mean = 516 CFU g-1). Significant differences were found in relation to both the investigated regions (p < 0.01) and the soil pH (r = 0.44*) (P = 0.02). The mycobacteria represented less than 0.00001% of the total aerobic bacteria found in the soils. Twenty-seven isolated mycobacteria were classified according to the culture, biochemical, enzymatic characteristics and antibiotic sensitivity. Mycobacterium fortitium was the dominant mycobacterium and was detected in 63% of the positive soils. This species showed ability for living in sandy to sandy loam soils, within a wide pH range (6.5-9.7) and organic matter (4.15-83.63 g kg-1). Two other species were M. phlei (range = 50-4,500 CFU g-1) and M. kansasii (range = 50-500 CFU g-1).

Nontuberculous mycobacteria in soils of La Pampa province (Argentina)

The presence of nontuberculous mycobacteria (NTM) was investigated in forty soil samples belonging to the four physiographic regions (Eastern, Central, Southern and Western) that constitute La Pampa province. The presence of NTM in 67.5 of these soil samples was determined. The density of mycobacteria ranged 25-4,500 mycobacteria g-1 dry soil (mean = 516 CFU g-1). Significant differences were found in relation to both the investigated regions (p < 0.01) and the soil pH (r = 0.44*) (P = 0.02). The mycobacteria represented less than 0.00001 of the total aerobic bacteria found in the soils. Twenty-seven isolated mycobacteria were classified according to the culture, biochemical, enzymatic characteristics and antibiotic sensitivity. Mycobacterium fortitium was the dominant mycobacterium and was detected in 63 of the positive soils. This species showed ability for living in sandy to sandy loam soils, within a wide pH range (6.5-9.7) and organic matter (4.15-83.63 g kg-1). Two other species were M. phlei (range = 50-4,500 CFU g-1) and M. kansasii (range = 50-500 CFU g-1).(AU)

Analysis of mycolic acids by high-performance liquid chromatography and fluorimetric detection. Implications for the identification of mycobacteria in clinical samples.

Mycolic acids from Mycobacterium phlei and M. bovis cell wall skeletons (CWSs) were analyzed by HPLC. After saponifying lyophilized CWSs in methanolic KOH, the mycolic acids were quantitatively extracted into chloroform. Aliquots of the CWS mycolic acid extracts were then derivatized prior to HPLC analysis with a UV reagent, p-bromophenacylbromide (PBPB), and three fluorescent reagents, 4-bromomethyl-6,7-dimethoxycoumarin, 4-bromomethyl-7-acetoxycoumarin and 3-bromomethyl-7-methoxy-1,4-benzoxazin-2-one. A synthetic alpha-branched carboxylic acid was derivatized with the same reagents and used as an internal standard along with the mycolic acids. The derivatized samples were analyzed by reversed-phase HPLC on a Waters Novapak C18, 4 microns particle size, 150 mm x 3.9 mm stainless-steel column. Two solvent systems were used: (1) methanol and methylene chloride with the column at 30 degrees C, and (2) methanol and isopropanol with the column at 50 degrees C. Detection sensitivity with the fluorescent reagents was 16-50 times greater than the sensitivity observed with PBPB-derivatized samples. Unique mycolic acid elution profiles for the two mycobacterial species could be achieved with each of the solvent systems and derivatization reagents tested. Thus, the HPLC analysis of pre-column derivatized mycolic acids was useful as a means of rapidly identifying mycobacterial species. Replacement of methylene chloride with isopropanol and PBPB with a fluorescent derivatizing reagent could increase the safety and sensitivity of the assay, and make it more useful for the clinical identification of mycobacterial infections.

Mycobacterium phlei infection of the foot: a case report.

A case of Mycobacterium phlei infection in the flexor digitorum longus and posterior tibialis tendon of an otherwise healthy adult male is reported. To our knowledge, this is the second reported case in the English literature of human infection by M. phlei. Diagnosis and treatment of nontuberculous mycobacterial infection are discussed. The clinician is encouraged to include this infection in the differential diagnosis of pain and swelling in the extremity.

Water soluble palmitic acid in media for cultivation of leprosy derived psychrophilic mycobacteria from Mycobacterium leprae infected tissues

Cultivation trials for Mycobacterium leprae resulted in growth of Mycobacterium psychrophilum (L). Media were inoculated with host grown Mycobacterium leprae cells from armadillo tissues, Nu mice foot pads or human lepromata. Cultures were obtained in liquid and on semisolid multifactoria 1 media containing water soluble palmitic acid or its salts. Ammonium thioglycolate and Napalmitate served as carbon and energy sources. The water soluble palmitic acid remained in perfect solution following sterilization in the autoclave, thus easily accessible to the cells. The cyclodextrin-Fe complex served as a siderophore to grow the obtained leprosy derived psychrophilic cells. The leprosy derived cultures and subcultures grew opimally at+10 degrees Celssius but deteriorated rapidly at + 32 degrees Celsius, in the multifactorial media. No growth occurred in 7H9 media. Cultures were not identified for classification.

[Identification of cultures of Mycobacteria cultivated using sodium salicylate medium]. / Identifikatsiia kul'tur mikobakterii, vyrosshikh na srede s salitsilatom natriia.

Sodium salicylate test is used in bacteriological practice to distinguish M. tuberculosis and M. bovis from other mycobacterial species. The species of the strains growing in sodium salicylate media are not as a rule identified. This was done using gas chromatography with cultures which had grown in Lowenstein-Jensen media with sodium salicylate and isolated from the patients admitted to the Tuberculous hospital in 1991-1993. Only 20% of the strains belonged to human mycobacteria, the others were opportunistic or saprophyte mycobacteria (M. avium, M. fortuitum, M. phlei, M. flavescens, M. vaccae, M. smegmatis), some of the cultures were not of the Mycobacterium genus, but belonged to other Actinomycetales. It is evident that with sodium salicylate test one cannot be absolutely sure of Mycobacteria isolation. The strains growing in sodium salicylate medium call for further investigations.

[Species category of Mycobacteria isolated from cattle and environmental objects]. / Vidovaia prinadlezhnost' mikobakterii, vydeliaemykh ot krupnogo rogatovo skota i iz ob"ektov vneshnei sredy.

The results of testing the slaughtered cattle material and environment objects for the presence of Mycobacteria are presented. During 1984-1988 with a stable excretion of pathogenic Mycobacteria, the quantity of the isolated atypical Mycobacteria tended to increase. In 1984 the atypical Mycobacteria made up 24.1% of the cultures isolated from cattle (pathogenic ones being 75.9%); in 1985, 29.0%; in 1986, 31.4%; in 1987, 46.0% and in 1988, 55.8%. For the above period M. bovis amounted to an average of 98.8% with annual fluctuations between 96.65 and 99.1%. During the last 5 years M. tuberculosis cultures ranged from 0.9 to 3.3% (1.92% on the average). M. avium was isolated from cattle in 22 cases out of 212 samples of the examined material (10.3%). As a result of testing of 2397 samples of the environment objects taken from 34 farms, 391 (16.3%) atypical Mycobacteria were isolated. It is necessary to continue, with regard to environmental conditions, the study of animal responses and atypical Mycobacterium carriage in different areas of the country.

Septic arthritis due to Mycobacterium phlei presenting as infantile Reiter's syndrome.

We describe a 7-year-old Peruvian boy who presented with a 2 week history of conjunctivitis, urethritis and arthritis, in whom Mycobacterium phlei was the only organism repeatedly isolated from synovial fluid and tissue and who responded to conventional antituberculous therapy. To our knowledge this is the first documented case of human disease caused by this microorganism.

Differentiation of Mycobacterium thermoresistibile from Mycobacterium phlei and other rapidly growing mycobacteria.

Mycobacterium thermoresistibile is the only chromogenic rapidly growing mycobacterial species reported to cause infections in humans and animals. DNA-DNA hybridization (S1 nuclease method) showed that M. thermoresistibile formed a DNA relatedness group which was only 24 to 30% related to M. phlei. Alkaline phosphatase and beta-galactosidase differentiated M. thermoresistibile from M. phlei. Among the rapidly growing mycobacteria, the mycolic acid pattern of M. thermoresistibile was unique (alpha-, alpha'-, methoxy-and keto-mycolates). Fourteen other species of chromogenic rapidly growing mycobacteria, including M. phlei, produced different mycolic acid patterns which always included dicarboxylic mycolate. Nine species of non-chromogenic rapidly growing mycobacteria produced mycolic acid patterns devoid of ketomycolate.

Search for acid-fast bacilli in bottled mineral waters.

Samples of bottled mineral water sold in Italy in accordance with the law on microbiological standards were examined for the presence of acid-fast bacilli. Eighty-four samples were tested, 11 with added carbon dioxide and 73 without. Acid-fast bacilli were found in 1 of the former samples and in 8 of the latter, a total of 10.7%. The mycobacterial count was always very low (2-3 cfu/litre) except for one sample which showed large numbers of Mycobacterium sphagni. This finding was not confirmed in two following samplings of the same water. The other acid-fast bacilli isolated were: M. gordonae (four strains), M. flavescens (one strain), M. phlei (one strain) and Nocardia sp. (two strains) in two samples of different water. These bacilli are not thought to be a normal component of the waters but are the result of incidental contamination. The risk of infection from drinking such waters can be regarded as irrelevant in hospitalized or immunologically compromised subjects who fact a greater risk when using tap water for drinking or washing.

Mycobacteremia and the new blood culture systems.

Clinical and microbiologic aspects of mycobacteremia occurring in 11 patients are presented. Ten of the 11 patients were immunosuppressed. Disseminated disease was shown in 7 patients, transient bacteremia in 2, and endocarditis in 1. Blood cultures were positive in 11 blood culture systems. Newer blood culture systems appear to provide more rapid detection of mycobacteremia.

[Atypical mycobacteria isolated from ganglions of presumed-healthy bovine and swine (author's transl)]. / Isolement de mycobactéries atypiques a partir de ganglions bovins et porcins présumés sains

A study has been carried out to detect the frequency of atypical mycobacteria in ganglions belonging to two types of animals: bovine and swine. A total of 250 samples (150 cows and 100 pigs) were studied. The samples were decontaminated by the lauryl-sulfate technique, using two mycobacteria culture media: Löwenstein-Jensen and Coletsos base. 14 pig ganglion strains were isolated (8 M. avium; 3 M. gordonae; 2 M. fortuitum; 1 M. chelonei) and 13 from the cow samples (5 M. avium, 3 M. Kansasii; 2 M. gordonae; 2 M. fortuitum; 1 M. phlei). The most frequently isolated mycobacteria was M. avium (48.14% of total isolated strains). This study confirms the possible importance of these animals in the epidemiology of mycobacteriosis.