Bacteriological features of Mycobacterium haemophilum isolated from skin lesions in an immunodeficient patient

Abstract

A 53-year-old, male patient presented with pain in the middle area of the back of his left foot. The painful area was associated with a reddish dome-shaped swelling of 24 by 18 mm which had ulcerated in the center part. Histopathologically, the cutaneous lesion consisted of an ulcer surrounded by abscess and granuloma and numerous acid-fast organisms were observed. Subsequently, the area just below the left inguinal area developed redness and swelling approaching the size of a quail egg. The patient responded favorably with rifampicin, levofloxacin, and minocycline therapy. The patient was immunodeficient, but negative for HIV-1 and HIV-2 antibodies and the etiology of his immunodeficient state is unclear. Skin tissues or pus were cultured at 37 degrees C on 2% Ogawa and BBL MGIT. Acid-fast organisms were recovered on MGIT within 4 to 12 days, while 2% Ogawa medium failed to recover acid-fast bacteria. Using growth from the positive MGIT tube as inoculum, MycoBroth, 7H9 broth, 7H11.2% Ogawa supplemented with or without iron complexes, and blood agar were inoculated and cultured at 30 and 37 degrees C. Growth at 30 and 37 degrees C was seen with MycoBroth, 7H9, hemin (60 microM) or ferric ammonium citrate (15 mg/ml) supplemented 7H11 and blood agar as well as 7H11 supplemented with factor X. Growth at 30 degrees C only was observed for ferric ammonium citrate supplemented 7H9 and 2% Ogawa. Generally, growth at 30 degrees C was better than that at 37 degrees C in all media. No growth at either temperature was observed with hemin or factor X supplemented 2% Ogawa. With respect to the biochemical characterization, the isolate was negative for niacin, nitrate reduction, urease, arylsulfatase, Tween 80 hydrolysis, catalase, 68 degrees C catalase, acid phosphatase, and tellurite reduction, while strongly positive for neutral red test. Sequencing of the 16S rRNA gene showed the isolate to be consistent with Mycobacterium haemophilum. Based on the composite characterization, the isolate was identified as M. haemophilum. This is the second case report of M. haemophilum infection in Japan in the literature.