Prevalence of non-tuberculosis mycobacteria among samples deposited from the National Tuberculous Reference Laboratory of Iran(2011-2018)

Objective:To investigate the prevalence of non-tuberculosis mycobacteria(NTM)among the samples deposited from the National Tuberculosis Reference Laboratory of Iran between 2011 and 2018.Methods:The study evaluated the prevalence of NTM among specimens from patients with pulmonary tuberculosis symptoms(n=15771)deposited at the National Tuberculosis Reference Laboratory of Iran from 2011 to 2018.Detection of Mycobacterium(M.)tuberculosis was based on presence of a 190-bp amplicon from IS6110 insertion sequence using Tb1 and Tb2 primers,and amplicon-negative specimens were tested for NTM and M.tuberculosis(refractory to IS6110 amplification)using restriction fragment length polymorphism PCR of hsp65 amplicon fragment.Results:A total of 7307(46.33%)M.tuberculosis and 658(4.17%)NTM specimens were found,the latter mainly comprising M.abscessus(10.18%),M.avium(2.28%),M.chelonae(8.97%),M.intracellulare(10.49%),M.kansasii(4.71%),and M.simiae(56.08%).Conclusions:As treatment for NTM differs from that for M.tuberculosis,accurate detection of Mycobacterium sp.is of public health significance.

First Report in China on the Identification and Drug Sensitivity of Mycobacterium elephantis Isolated from the Milk of a Cow with Mastitis

Objective In this study, milk from a cow with mastitis was analyzed to determine the presence of mycobacterial infection. Milk quality and security problems pertaining to the safe consumption of dairy products were also discussed in this study. Methods Milk was preprocessed with 4% NaOH. Then, mycobacteria were isolated from the milk sample on L-J medium. The isolate was identified using multiple loci Polymerase Chain Reaction （PCR） and multi-locus sequence analysis with 16S rRNA, sodA, hsp65, and ITS genes. The drug sensitivity of the isolate to 27 antibiotics was tested through alamar blue assay. Results Smooth, moist, pale yellow colonies appeared on the L-J medium within a week after inoculation. Based on the results of multiple loci PCR analysis, the isolate was preliminarily identified as non-tuberculous mycobacteria. The 16S rRNA, SodA, hsp65, and ITS gene sequences of the isolate exhibited 99%, 99%, 99%, and 100% similarities, respectively, with those of the published reference strains of Mycobacteriurn elephantis （M. elephantis）. The drug sensitivity results showed that the strain is resistant to isoniazid, p-aminosalicylic acid, and trimesulf but is sensitive to ofloxacin, rifampicin, amikacin, capreomycin, moxifloxacin, kanamycin, levofloxacin, cycloserine, ethambutol, streptomycin, tobramycin, rifabutin, ciprofloxacin, linezolid, cefoxitin, clarithromycin, and minocycline. Conclusion To the best of our knowledge, this study is initially to report the isolation of M. elephantis from the milk of a cow with mastitis in China.

Pathological humerus fracture due to anti-interferon-gamma autoantibodies:A case report

BACKGROUND Various etiologies contribute to pathological fractures,including bone infections.Recently,non-tuberculosis Mycobacterium-related bone infections among patients with anti-interferon-gamma autoantibody-induced adult-onset immunodeficiency has raised concerns in Southeast Asia,with the common presentations including osteomyelitis.However,it also rarely manifests as traumatic fractures,as reported in this case.CASE SUMMARY A diabetic female fractured her humerus after a traumatic accident and received fixation surgery.Abnormal necrotic bone tissue and abscess formation were noted,and she was diagnosed with a pathological fracture due to nontuberculosis Mycobacterium infection.Multiple bone involvement was also revealed in a bone scan.Anti-interferon-gamma autoantibodies were then checked due to an unexplained immunocompromised status and found to be positive.Her humerus fracture and multiple bone infections healed after steroid and anti-non-tuberculosis Mycobacterium medication treatment following fixation surgery.CONCLUSION Comprehensive preoperative evaluations may help identify pathological fractures and guide the treatment course.

Characterization of Mycobacterium Species Circulating among Tuberculosis Patients in Bayelsa State, Nigeria

Introduction: Tuberculosis is caused by infection with Mycobacterium tuberculosis. Looking at the evolution of the bacterium gene due to mutation is crucial to identify species circulating among patients in an area. WHO speculated that tuberculosis is caused by M. tuberculosis (MTB), but identification of the strains of MTB circulating in a particular area is important for the management of MTB and to identify pulmonary infections caused by non-tuberculosis mycobacterium. Contact tracing of drug resistant MTB in circulation in an area is also an important procedure of MTB therapeutic choice. Aim: This study aimed to isolate and identify Mycobacterium species circulating in Bayelsa State, Nigeria. Materials and Methods: A total of 102 sputum samples collected from MTB patients were cultured in Lowenstein Jensen (LJ) solid media. Isolates on LJ media were confirmed using Zeihl Nelseen staining method for AFB and Standard Diagnosis Bioline TB Ag MPT64 Rapid test kit. The 16s rRNA gene amplification, agarose gel electrophoresis, and gene sequencing were conducted. Phylogenic analysis and evolutional distances of the strains are computed using the Juke-cantor method. Result: Out of 102 sputum samples examined 15 (14.7%) had growth of Mycobacterium species (AFB positive). The extracted DNA of MTB amplified on agarose gel electrophoresis aligned horizontally at lanes 1 - 15 showing 16S gene band (1500 bp). Two 2 (2.0%) are non-tuberculosis Mycobacteria species, while 13 (12.7%) were M. tuberculosis. The non-tuberculosis Mycobacterium species isolated are Mycobacteriode abscesses and Mycobacterium kansassii strain FDAARGOS 1534. The tuberculosis strains are Mycobacterium tuberculosis MG003 and R2092 but the predominant strain was MG003. The degree of the genetic evolution of the non-MTB Mycobacterium kansassii strain FDAARGOS 1534 was 75.4%. Conclusion: The two major strains of Mycobacterium tuberculosis (MTB) circulating in Bayelsa State are MTB MG003 and MTB R2092;MTB MG003 was predominant. The non-tuberculosis species are Mycobacteriode abscesses and Mycobacterium kansasii.

Genitourinary Bacillus Calmette-Guerin Infection after BCG Intravesical Administration in a Patient under Hemodialysis—What Can We Do Better?

A 74-year-old man with terminal chronic kidney disease, under hemodialysis and with residual diuresis, was admitted due to myalgia, arthralgia, fever and pyuria in the previous 10 days. The patient had a recent diagnosis of high-grade non-invasive bladder cancer and was doing weekly BCG intravesical administrations. The symptoms started three days before the fifth administration. He had done cefixime as an outpatient and started piperacillin-tazobactam on hospital admission, but the fever persisted, and there was no bacterial isolation in urine or blood culture. On the tenth and seventeenth day after the last BCG intravesical administration Mycobacterium bovis was still isolated in the urine culture. The diagnosis of BCGitis was made and treatment was started, with a good response. Forty days after the last administration and under treatment, the culture remained positive for Mycobacterium bovis in the urine. We raise the question about the safety of BCG administration in patients with residual diuresis.

Saccharomyces boulardii Produces a Factor That Inhibits Mycobacterium intracellulare Burden in Human Macrophages

Lung disease caused by Non-Tuberculosis Mycobacteria (NTM) is increasing in prevalence. NTM lung disease is notable for poor response to therapy. Saccharomyces boulardii is probiotic that can be effective in inflammatory gastrointestinal disease with diverse pathophysiology. The present study investigated the effects of the products of S. boulardii-B508 on burden of NTM-Mycobacterium intracellulare complex in human macrophage infection in vitro. It was found that the supernatant of S. boulardii-B508 inhibited the growth of Mycobacterium intracellulare in human macrophage infection and induced infected cell apoptosis. The data of RT-PCR showed that the products of S. boulardii-B508 inhibited IL-8 expression during M. intracellulare infection in human macrophages due to its effects on NF-kB activation. To the best of our knowledge, this is the first report of effective products of S. boulardii on NTM infection in human macrophage. S. boulardii possesses anti-NTM lung disease properties in human macrophage worthy of further evaluation in clinical studies.