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The list of clinically important nontuberculous mycobacteria (NTM) is growing as new species continue to be identified and older ones are found to be pathogenic. Molecular techniques such as real-time PCR and gene amplification and restriction length polymorphism are promising tools for rapid identification of NTM. Optimal therapy for a documented Mycobacterium gordonae infection remains undefined. The majority of isolates tested have been resistant in vitro to isoniazid and pyrazinamide, whereas many are susceptible to ethambutol, rifampin, clarithromycin, linezolid, and the fluoroquinolones. Unlike other nontuberculous mycobacteria, M. simiae produces niacin and thus may be confused with M. tuberculosis. M. simiae can colonize the respiratory tract, and the lung is the most commonly reported site of infection. The majority of reported infections have occurred in persons living near rivers or stagnant bodies of water. A recent open-label randomized trial conducted in Ghana compared two regimens for early, limited M. ulcerans infections. In this study, 73 of 76 patients who received streptomycin and rifampin for 8 weeks and 68 of 75 patients who received 4 weeks of streptomycin and rifampin followed by 4 weeks of rifampin and clarithromycin had healed lesions at 1 year after the start of treatment. Infections with M. xenopi have shown variable responses to drug therapy. Recommendations for initial therapy include isoniazid, a rifamycin, ethambutol, and clarithromycin with or without an initial course of streptomycin. Pyrazinamide and ciprofloxacin have been included in some successful regimens.

Background/purpose Mycobacterium gordonae is a ubiquitous environmental mycobacteria and has been long considered an opportunistic pathogen, causing infections only in immunocompromised hosts. Cases of M. gordonae related infections in immunocompetent host have rarely been reported, and the pathogenicity of M. gordonae remained uncertain. Methods From January 2016 to December 2018, seven cases of M. gordonae infection were diagnosed and treated at National Taiwan University Hospital. Results Six cases had at least one underlying disease affecting immune status, while one case had no identifiable underlying disease. The sites of infection were lung (n = 3), skin and soft tissue (n = 3), and one had disseminated disease involving the lung and bone marrow. All patients were cured after anti-mycobacterial treatment, except one patient died of refractory leukemia. Conclusion Compatible with the literature reports, we demonstrate that M. gordonae could be pathogenic and causing infection not only in the immunocompromised host, but also in the otherwise healthy population. Multi-antimicrobial combination and adequate source control could have good therapeutic effect for patients with M. gordonae infections

Tuberculosis (TB) continues to pose a significant public health problem worldwide. For mycobacteriology laboratories, it is important to be able to diagnose active cases and to make a differential diagnosis of non-tuberculous mycobacteria (NTM). In this study, it was aimed to retrospectively evaluate the epidemiological status of the Mycobacterium [Mycobacterium tuberculosis complex (MTC) and NTM] obtained from the clinical specimens of patients with TB suspicion, and the resistance rates of MTC isolates against anti-TB drugs. Various clinical samples of TB suspected patients sent to the Balikesir Ataturk City Hospital Mycobacteriology Laboratory between 2011 and 2019, were included in the study. Microscopy, culture procedures, and the first-line anti-TB drug susceptibility tests were performed according to the instructions. Identification of NTM at the species level could be made during four years including 2012, 2013, 2016, and 2017. In our study, acid fast bacillus (AFB) positivity rate was 4% (1.867/47.235); the culture positivity rate for MTC was 5.1% (1.576/31.017) and 1.1% (333/31.017) for NTM. AFB positivity was detected in 837 (53.1%) of the clinical specimens isolated from MTC. In the presence of AFB positivity, it was determined that bacterial growth was significantly higher in both liquid culture systems (LCS) and Lowenstein-Jensen (LJ) media. The isolation rate of MTC isolates from LCS was determined as 95.3% (1.503/1.576) and the isolation rate from LJ was 67.4% (1.063/1.576). The bacterial growth rate was found to be significantly higher in LCS. The average bacterial growth time (ABGT) of AFB negative samples were 21.79 ± 9.96 days; 13.74 ± 8.13 days for AFB positive samples, and ABGT was significantly shorter in the case of AFB positivity. As the severity of AFB positivity increased, ABGT wasshortened which was statistically significant. While 783 (78%) of the isolates were found to be sensitive to all the tested drugs, 221 (22%) were found to be resistant to at least one drug. Eleven of them (1%) were identified as multidrug resistant-TB (MDR-TB) isolates. In our study, 16 different species were identified among 112 typed NTM isolates. Mycobacterium gordonae (25.0%), Mycobacterium avium complex (17.0%) (Mycobacterium intracellulare-11.6%, Mycobacterium avium-5.4%) and Mycobacterium abscessus (13.3%) were the most frequently isolated NTM species. As a result, nine-year results of the mycobacteriology laboratory in our region were analyzed and the MTC and NTM epidemiological data were determined for the first time.

In this study, mycobacteria, which were previously identified as Mycobacterium tuberculosis complex (MTC), and mycobacteria other than tuberculosis (MOTT) with cord factor and the p‐nitro‐alpha‐acetyl‐amino‐beta‐hydroxypropiophenone (NAP) test were reanalysed using the polymerase chain reaction—restriction fragment length polymorphism (PCR‐RFLP) analysis method in order to confirm the identification, and at the same time, species accepted as MOTT were identified. Although the results of the NAP test were obtained within 3‐5 days, the PCR‐RFLP results were obtained in 1 day. Ten species identified as MTC with the NAP test and cord factor were confirmed with the PCR‐RFLP method. Fourteen species accepted as MOTT were identified as Mycobacterium species with the evaluation of the bands observed after the restriction of PCR product with the PCR‐RFLP method. These were as follows: three species Mycobacterium intracellulare type I, two species Mycobacterium phlei, two species Mycobacterium kansasii, one species Mycobacterium fortuitum type I, one species Mycobacterium gordonae type I, one species Mycobacterium abscessus type I, one species Mycobacterium scrofulaceum, one species Mycobacterium szulgai type I, one species Mycobacterium avium type II, and one species Mycobacterium terrae. Hence, the results of both the cord factor and the NAP test were confirmed with the molecular method, and at the same time, mycobacteria species identification was made by determining the fastest, easiest, and the most accurate result‐giving method. Because PCR‐RFLP is a very rapid method that provides exact identification of mycobacteria species, it can be performed in routine procedures.

L-asparaginase (E.C.3.5.1.1) is a well-known agent that prevents the formation of acrylamide both in the food industry and against childhood acute lymphoblastic leukemia in clinical settings. The disadvantages of L-asparaginase, which restrict its industrial application, include its narrow range of pH stability and low thermostability. In this study, a novel L-asparaginase from Mycobacterium gordonae (GmASNase) was cloned and expressed in Escherichia coli BL21 (DE3). GmASNase was found to be a tetramer with a monomeric size of 32 kDa, sharing only 32% structural identity with Helicobacter pylori L-asparaginases in the Protein Data Bank database. The purified GmASNase had the highest specific activity of 486.65 IU mg−1 at pH 9.0 and 50 °C. In addition, GmASNase possessed superior properties in terms of stability at a wide pH range of 5.0–11.0 and activity at temperatures below 40 °C. Moreover, GmASNase displayed high substrate specificity towards L-asparagine with Km, kcat, and kcat/Km values of 6.025 mM, 11,864.71 min−1 and 1969.25 mM−1min−1, respectively. To evaluate its ability to mitigate acrylamide, GmASNase was used to treat potato chips prior to frying, where the acrylamide content decreased by 65.09% compared with the untreated control. These results suggest that GmASNase is a potential candidate for applications in the food industry.

Nontuberculous mycobacteria (NTM) are environmental opportunistic pathogens of humans and animals that are emerging with a serious public health impact particularly in individuals with Acquired Immunodeficiency Syndromes. Recent scientific evidence is shifting from NTMs being known as traditional environmental organisms to serious pathogenic organisms in both animals and humans. In humans, factors attributable to this rise have been linked mainly to Human Immunodeficiency Virus and Acquired Immunodeficiency Syndrome pandemic. In recent years there has been an increase in multidrug resistant Tuberculosis in Zambia and it is thought that NTMs could possibly be the cause. This study was therefore formulated to review available information on the prevalence of NTM in humans, animals and the environment, species distribution, zoonotic potential and public health importance in Zambia. This review was conducted in accordance with PRISMA guidelines. A literature search was done in PubMed and Google scholar using predefined search terms such as ‘nontuberculous mycobacteria’, ‘atypical mycobacteria’, ‘mycobacteria other than mycobacterium tuberculosis’ and ‘Zambia’, in combination with Boolean operators (AND, OR). This particular systematic review draws findings based on literature search between 2000 and 2020. Through literature search 243 papers were identified, 23 duplicates were identified and removed and 206 articles were excluded as they did not meet the inclusion criteria. The full text of the remaining 14 articles were considered for this review. The overall prevalence of NTM in humans was 24.39%, in water 21.5%, in animals 16.05% of which the prevalence in cattle was 14.81% and Kafue Lechwe 1.23%. Mycobacterium intracellulare was the most common isolated nontuberculous mycobacteria in humans and cattle while Mycobacterium gordonae was the most common in water, and Mycobacterium stomatepiae sp Nov in Kafue Lechwe. Nontuberculous mycobacteria are an emerging public health threat in Zambia both in humans and animals and this calls for the need for molecular information on the zoonotic transmission of nontuberculous mycobacteria. Increased awareness of nontuberculous mycobacteria diseases among clinicians and laboratory personnel is crucial for patient management and an essential step for facilitating the identification of nontuberculous mycobacteria species in laboratories.

Made available in DSpace on 2022-04-28T17:22:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundacao Cearense de Apoio ao Desenvolvimento Cientifico e Tecnologico - FUNCAP Emerging infectious diseases in wild animals related to humans have received greater attention in recent years. Mycobacteriosis is a bacterial disease of animal and human importance. Mycobacterium gordonae infects the skin and internal organs of free-ranging amphibians and is considered the least pathogenic member of the Mycobacteriaceae to humans. However, information about its infection and pathogenesis in wild amphibians is still lacking. A total of 1306 amphibian specimens belonging to 6 families, 12 genera, and 21 species were collected and dissected during a helminthological survey of 7 municipalities in southern Ceara state, Caatinga (eco)region, northeast Brazil. Of these, 17 specimens (0.76%), belonging to 2 families and 4 species (Leptodactylus macrosternum, n = 2; L. vastus, n = 10; Pseudopaludicola pocoto, n = 2; Rhinella jimi, n = 3), presented infections that consisted of calcification nodules in the coelomic cavity, kidney, liver, lung, gut, and pancreas. The nodules were examined by histopathology and PCR. The bacteria were identified as M. gordonae by molecular analyses. Infected animals presented with hepatocellular vacuolar degeneration, karyolysis, and karyorrhexis, hepatic portal congestion, hemorrhage, mononuclear cellular infiltration, melanomacrophage center hyperplasia, and granulomas in varying stages of development with intralesional acid-fast bacilli. This study is the first report of M. gordonae in these amphibian species, in which results of molecular analyses confirmed the presence of M. gordonae in natural environments and histopathology confirmed the typical lesion of mycobacteriosis in amphibians from northeastern Brazil. Univ Fed Uberlandia UFU, Inst Ciencias Agr, BR-38500000 Monte Carmelo, MG, Brazil Univ Estadual Paulista UNESP, Dept Parasitol, Inst Biociencias, BR-18618970 Botucatu, SP, Brazil Univ Fed Ceara UFC, Ctr Reg Ophiol, BR-60455760 Fortaleza, Ceara, Brazil Univ Estadual Paulista UNESP, Dept Parasitol, Inst Biociencias, BR-18618970 Botucatu, SP, Brazil CNPq: 303622/2015-6 CNPq: 305988/2018-2 CNPq: 313241/2018-0 Fundacao Cearense de Apoio ao Desenvolvimento Cientifico e Tecnologico - FUNCAP: AEP-0128-00269.01.00/17 Fundacao Cearense de Apoio ao Desenvolvimento Cientifico e Tecnologico - FUNCAP: 05/2017

BACKGROUND Nontuberculous mycobacteria (NTM) species are a rising threat, especially to patients living with pulmonary comorbidities. Current point-of-care diagnostics fail to adequately identify and differentiate NTM species from Mycobacterium tuberculosis (Mtb). Definitive culture- and molecular-based testing can take weeks to months and requires sending samples out to specialized diagnostic laboratories. METHODS In this proof-of-concept study, we developed an assay based on PCR amplification of 16S ribosomal RNA (rRNA) rrs genes by using universal mycobacterial primers and interrogation of the amplified fragments with a panel of binary deoxyribozyme (BiDz) sensors to enable species-level identification of NTM (BiDz-NTMST). Each BiDz sensor consists of 2 subunits of an RNA-cleaving deoxyribozyme, which form an active deoxyribozyme catalytic core only in the presence of the complimentary target sequence. The target-activated BiDz catalyzes cleavage of a reporter substrate, thus triggering either fluorescent or colorimetric (visually observed) signal depending on the substrate used. The panel included BiDz sensors for differentiation of 6 clinically relevant NTM species (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium fortuitum, Mycobacterium kansasii, and Mycobacterium gordonae) and Mtb. RESULTS Using the fluorescent BiDz-NTMST assay, we successfully identified the species of 38 clinical isolates. In addition, a subset of strains was tested with visual BiDz sensors, providing proof-of-concept for species typing of NTM by the naked eye. CONCLUSIONS The BiDz-NTMST assay is a novel platform for rapid identification of NTM species. This method is highly specific and significantly faster than current tools and is easily adaptable for onsite diagnostic laboratories in hospitals or clinical laboratories.

Background Nontuberculous mycobacterium (NTM) infection in a patient in an immunosuppressed state caused by increased use of immunosuppressive or biological agents is a serious clinical problem. Mycobacterium avium complex is the most common involved pathogen, followed by Mycobacterium abscessus (MABSC), while Mycobacterium kansasii is not a major concern. The rate of infection rate in lung transplant recipients is reported to range from 1.5–22.4%. Case presentation We report here four cases of NTM pulmonary infection and disease among 63 patients who underwent lung or heart-lung transplantation at our hospital. Those four occurred following living-donor transplantation in two patients, one with pulmonary arterial hypertension and one with bronchiectasia, and deceased donor lung transplantation in two patients, lymphangioleiomyomatosis and interstitial pneumonia, respectively. NTM was not detected in any of the patients prior to transplantation. The involved pathogens were Mycobacterium gordonae in one, MAC in one, and MABSC in two of these patients, which were isolated from broncho-alveolar lavage (BAL) in two and sputum in two. The one case of MAC and two of MABSC were symptomatic with consolidation shown in chest CT images indicating possible pneumonia, while the one with M. gordonae had no symptoms and was detected by surveillance BAL. Onset time from detection of NTM was greater than 3 years in the three with MABSC and M. gordonae and less than 3 years in the one with MAC. Each patient required a decrease in immunosuppressive agents according to their condition, while antibiotics therapy was performed in the three who were symptomatic. Sputum culture findings became negative after several months and were maintained thereafter in all. Conclusion An NTM infection leading to pulmonary disease can occur at any time following lung transplantation. Treatment should be considered depending on the involved pathogens, individual status, and disease severity.

In this study, we investigated the immune responses against Mycobacterium gordonae in ginbuna crucian carp. Cumulative mortality of ginbuna injected with 2.0 × 107 CFU of M. gordonae was 50% at 170 days post-infection. CD4-1, CD8α, T-bet and IFNγ2 gene expression levels were significantly upregulated in ginbuna injected with 1.9 × 108 CFU of M. gordonae at 21 and 28 days post-infection. The CD4-2 level did not change during the experiment. Granulomatous responses consisted of central macrophage accumulation and surrounding lymphocytes, and Ziehl-Neelsen-positive bacteria were observed in the trunk kidney of the challenged fish. Immunohistochemistry using anti-ginbuna IFNγs and anti-ginbuna CD4-1 polyclonal antibody revealed that the marginal lymphocytes were positive for CD4-1, and the IFNγ-producing cells surrounded the mycobacterial cell-laden phagocytes. These results suggest that CD4-1+ cells and IFNγ2 play important roles in the granulomatous inflammation against Mycobacterial infections in teleosts.

Background: Nontuberculous mycobacteria (NTM) have been identified in human pulmonary and extrapulmonary infections and are increasing globally, which makes it challenging to identify them. This article reports our experience with the laboratory identification of NTM in clinical practice among pulmonary and extrapulmonary samples received in our routine work. Methods: The study was conducted at the Universite Catholique de Louvain at the Cliniques Universitaires Saint-Luc, Brussels, Belgium, from 2015 to 2018. A total of 386 clinical samples were collected from patients suspected of having pulmonary or extrapulmonary mycobacterial infections. Routine laboratory methods phenotypic and molecular tests were performed. Results: The majority of NTM species were isolated from pulmonary samples (68%). The most prevalent species identified were Mycobacterium chimaera_intracellulare group (32%), followed by Mycobacterium avium complex (21%), Mycobacterium abscessus complex (18%), Mycobacterium gordonae (9%), and Mycobacterium chelonae (4%). In extrapulmonary samples, M. avium and M. chimaera_intracellulare were the most frequently isolated. Conclusion: The species diversity of NTM found in our setting suggests the importance of the use of new modern methods for accurate identification of NTM at species level and in some case at subspecies level for the proper treatment and management of patients.

A slow-growing, scotochromogenic mycobacterial strain (24T) was isolated from the sputum of a Chinese male human. Phylogenetic analysis using the 16S rRNA gene assigned strain 24T to the Mycobacterium gordonae complex, which includes Mycobacterium gordonae and Mycobacterium paragordonae . The phenotypic characteristics, unique mycolic acid profile and the results of phylogenetic analysis based on hsp65 and rpoB sequences strongly supported the taxonomic status of strain 24T as a representative of a species distinct from the other members of the M. gordonae complex. The genomic G+C content of strain 24T was 65.40mol%. Genomic comparisons showed that strain 24T and M. gordonae ATCC 14470T had an average nucleotide identity (ANI) value of 81.00 % and a DNA–DNA hybridization (DDH) value of 22.80 %, while the ANI and DDH values between strain 24Tand M. paragordonae 49 061T were 80.98 and 22.80 %, respectively. In terms of phylogenetic, phenotypic and chemotaxonomic features, strain 24T is distinguishable from its closest phylogenetic relatives and represents a novel species of the genus Mycobacterium , therefore the name Mycobacterium vicinigordonae sp. nov. is proposed. The type strain is 24T (=CMCC 93559T=DSM 105979T).

A 46-year-old man visited our hospital with a fever and cough. The symptoms had started two months after continued use of an ultrasonic humidifier. He had hypoxemia on admission and late inspiratory crackles in both lungs on physical examination. The laboratory findings showed an increased white blood cell count and a C-reactive protein level, and his serum KL-6 level was slightly elevated, at 674 U/mL. Chest computed tomography showed diffuse ground-glass opacities, and histological examination of a transbronchial lung biopsy showed alveolitis without granulomas. The humidifier inhalation challenge test result was positive. Therefore, we diagnosed the patient with humidifier lung. His symptoms gradually improved after avoiding the humidifier without taking medication. The humidifier water was contaminated by various bacteria and fungi, as well as Mycobacterium gordonae and a high concentration of endotoxin. Unlike in those with typical hypersensitivity pneumonitis, the elevation of serum KL-6 levels in humidifier lung patients is mild, and granulomas are not apparent on histological examination, similar to our case. Furthermore, the endotoxin identified from the humidifier is one of the known pathogens of humidifier lung. Thus, humidifier lung seems to have different characteristics compared to other hypersensitivity pneumonitis phenotypes. The mechanism driven by the high concentration of endotoxin could be one of the main causes of humidifier lung.

Mastitis is a common disease in women with both infectious and noninfectious causes. Most cases occur during lactation and are caused by Staphylococcus aureus and Streptococcus species; parasites and Mycobacteria have rarely been reported to cause breast infections (Mandell, Douglas, and Bennett's principles and practice of infectious diseases (9th edn);2019, Am J Respir Crit Care Med. 2007;175:367). Nontuberculous mycobacteria (NTM) which are also referred to as atypical mycobacteria, mycobacteria other than tuberculosis (MOTT), or environmental mycobacteria are a large group of Mycobacteria which are becoming increasingly common cause of infection all over the world (Arch Dermatol. 2006;142:1287). NTM can cause infection diseases especially in immunocompromised patients, such as HIV-positive hosts, most commonly in the lungs, skin and soft tissue, lymph nodes or rarely spread with multiorgan dissemination (Arch Plast Surg. 2014;41:759). Mycobacterium gordonae (M. gordonae) is a slow-growing atypical mycobacterium that is considered the least pathogenic NTM. The organism is ubiquitous, and mostly isolated from soil and water. Despite its nonvirulent nature, clinically significant infections have been reported also in some immunocompetent patients (J Formosan Med Assoc. 2020, Clin Infect Dis. 1992;1229). We report the first documented case of breast infection in a young immunocompetent woman sustained by Mycobacterium Gordonae.

SUMMARY Objective The international outbreak of cardiac-surgery-associated Mycobacterium chimaera infections was traced back to infectious aerosols originating from contaminated water reservoirs of heater-cooler devices (HCDs). In general, non-tuberculous mycobacteria (NTM) frequently colonize water systems and can contaminate medical devices. Data on detection of NTM other than M. chimaera in samples gathered from HCDs are scarce. The present study summarizes prospective mycobacterial surveillance of five HCDs over more than 4 years. Methods Five LivaNova 3T (London, UK) HCDs, acquired factory-new in 2014, were followed prospectively. Until mid-April 2014, the HCDs were maintained according to the manufacturer's recommendations, and subsequently, they were maintained according to an intensified in-house protocol including exhaust air evacuation. Mycobacterial surveillance cultures consisted of monthly water samples gathered from patient and cardioplegia circuits, as well as airflow samples. Results Of 441 water samples, 170 (38.6%) revealed NTM growth. The most frequently detected NTM were M. chimaera [N=120 (67.4%)], Mycobacterium gordonae [N=35 (19.7 %)] and Mycobacterium paragordonae [N=17 (9.6%)]. Growth of NTM, M. chimaera and M. paragordonae was significantly more common in water samples derived from the patient circuit than the cardioplegia circuit of the HCD. Three (2.0%) of 150 air samples grew NTM. Conclusion Growth of NTM in HCD water samples was common. Diverse NTM species were detected, with M. chimaera being the most common. The majority of air samples remained negative. The relevance of NTM other than M. chimaera contaminating HCDs is poorly defined, but a recent report on an HCD-associated outbreak with Mycobacterium abscessus confirmed a potential threat.

Mycobacterium tuberculosis infection and nontuberculous mycobacteria (NTM) infections exhibit similar clinical symptoms; however, the therapies for these two types of infections are different. Therefore, the rapid and accurate identification of M. tuberculosis and NTM species is very important for the control of tuberculosis and NTM infections. In the present study, a Cas12a/guide RNA (gRNA)-based platform was developed to identify M. tuberculosis and most NTM species. By designing species-specific gRNA probes targeting the rpoB sequence, a Cas12a/gRNA-based platform successfully identified M. tuberculosis and six major NTM species (Mycobacterium abscessus, Mycobacterium intracellulare, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium gordonae, and Mycobacterium fortuitum) without cross-reactivity. In a blind assessment, a total of 72 out of 73 clinical Mycobacterium isolates were correctly identified, which is consistent with previous rpoB sequencing results. These results suggest that the Cas12a/gRNA-based platform is a promising tool for the rapid, accurate, and cost-effective identification of both M. tuberculosis and NTM species.

Acanthamoeba is an opportunistic protozoon, widespread in the aquatic environment, where it can be in endosymbiosis with over 30 pathogenic bacteria, including nontuberculous mycobacteria (NTM). Protozoa play a crucial role in mycobacterial pathogenesis and serve as a reservoir of infection. Since the first step in bacteria making contact with amoebae is adhesion, we were interested in investigating whether essential oils (EOs) can affect it. To that end we investigated the effects of juniper (Juniperus communis) and immortelle (Helichrysum italicum) EOs against Mycobacterium avium, M. intracellulare, and M. gordonae in tap water and against their adhesion to Acanthamoeba castellanii by combining them in synergistic EO concentrations. M. avium and M. intracellulare adhered to A. castellanii to a greater extent than M. gordonae. The adhesion of all NTMs was prevented by the subinhibitory concentrations of EOs. When comparing the effect of synergistic combinations of EOs and the effect of a single concentration from a combination, a higher percentage of adhesion inhibition in all synergistic combinations observed, except against M. gordonae. Neither oil was cytotoxic to A. castellanii. Our findings suggest that the EOs or their components weaken the contact of environmental NTMs and free-living amoebae and indirectly diminish their pathogenic potential, which could be of value in developing strategies for maintenance of water supply systems., Akantameba je oportunistička protozoa, široko rasprostranjena u vodenom okružju, gdje može kao endosimbionte sadržavati više od 30 različitih patogenih bakterija, uključujući netuberkulozne mikobakterije (NTM). Protozoe kao rezervoar infekcije imaju ključnu ulogu u mikobakterijskoj patogenezi. Sposobnost mikobakterija da se vežu, ulaze I umnožavaju se unutar protozoa vjerojatno im je omogućila da postanu intracelularni patogeni. Budući da je adhezija prvi korak u uspostavi kontakta bakterija s amebama, zanimalo nas je istražiti mogu li prirodni spojevi poput eteričnih ulja utjecati na taj kontakt. Cilj ove studije bio je istražiti učinak eteričnih ulja borovice (Juniperus communis) i smilja (Helichrysum italicum) prema NTM-ovima (M. avium, M. intracellulare i M. gordonae) u pitkoj vodi. Također, ispitana je mogućnost inhibicije adhezije tih NTM-ova na Acanthamoeba castellanii (A. castellanii) primjenom sinergijskih koncentracija eteričnih ulja. M. avium i M. intracellulare, u odnosu na M. gordonae, u većoj su mjeri adherirali na A. castellanii. Adhezija svih NTM-ova inhibirana je subinhibicijskim koncentracijama eteričnih ulja borovice i smilja. Kad se usporedi učinak sinergijskih kombinacija i učinka pojedine koncentracije iz sinergije, uočen je veći postotak inhibicije adhezije kod primjene svih sinergijskih kombinacija, osim jedne prema M. gordonae. Oba eterična ulja u ispitivanim koncentracijama nisu imala citotoksično djelovanje na stanice A. castellanii. Na taj su način primijenjene subinhibicijske koncentracije eteričnih ulja sa smanjenim toksičnim učinkom, uz zadržan antimikrobni i antiadhezivni učinak. Primjena eteričnih ulja ili njihovih komponenata u umjetnom vodenom okolišu mogla bi oslabiti kontakt NTM-ova i slobodno živućih ameba i posredno dovesti do smanjenja patogenoga potencijala okolišnih NTM-ova.

WOS: 000453790600009, PubMed: 30010392, The typing of non-tuberculous mycobacteria (NTM) is important from a clinical and epidemiological perspective. The polymerase chain reaction-restriction enzyme analysis (PRA) method and DNA sequence analysis method were utilized to target a gene region that codes the 65-kDa heat-shock protein for typing 150 suspected NTM samples isolated from the respiratory tract. Mycobacterium abscessus, Mycobacterium xenopi, Mycobacterium fortuitum, and Mycobacterium peregrinum were most frequently found by both methods. Six isolates that could not be defined by the PRA method were defined as Nocardia cyriacigeorgica, Nocardia abscessus, and Mycobacterium intracellulare by DNA sequence analysis. Discordance between the results of the two methods was observed for only one isolate. The isolate that was defined as Mycobacterium gordonae type 6 by the PRA method was defined as Mycobacterium senegalense by sequence analysis. The PRA method is simple and gives rapid results. Compared with DNA sequence analysis, it gives consistent and reliable results up to a ratio of 90%. DNA sequence analysis is the gold standard method in which all strains can be defined. However, given our laboratory conditions, its disadvantage is that it takes longer to reach a diagnosis than through the PRA method.

Background Ataxia telangiectasia (AT) is a rare, multi-systemic, genetic disorder. Mutations in the ATM gene cause dysfunction in cell-cycle, apoptosis and V (D) J recombination leading to neurodegeneration, cellular, humoral immunodeficiencies and predisposition to malignancies. Previous studies have suggested that a sub-group of AT patients with elevated IgM levels have a distinct and more severe phenotype. In the current study we aimed to better characterize this group of patients. Methods We performed a retrospective review of 46 patient records, followed from January 1986 to January 2015 at the Israeli National AT Center. Demographic, clinical, radiological, laboratory data was reviewed and compared between AT patients with elevated IgM levels (EIgM) and patients with normal IgM levels (NIgM). Results 15/46(32.6%) patients had significantly elevated IgM levels. This group had a unique phenotype characterized mainly by increased risk of infection and early mortality. Colonization of lower respiratory tract with Mycobacterium gordonae and Pseudomonas aeruginosa as well as viral skin infections were more frequent in EIgM patients. Patients with NIgM had a significantly longer survival as compared to patients with EIgM but had an increased incidence of fatty liver or cirrhosis. T-cell recombination excision circles and kappa-deleting element recombination circle levels were significantly lower in the EIgM group, suggesting an abnormal class switching in this group. Conclusions EIgM in AT patients are indicative of a more severe phenotype that probably results from a specific immune dysfunction. EIgM in AT should be considered a unique AT phenotype that may require different management. Electronic supplementary material The online version of this article (10.1186/s12887-018-1156-1) contains supplementary material, which is available to authorized users.

Previous studies have identified patients infected with Mycobacterium chimaera (M. chimaera) subsequent to cardiac surgery. Water tanks in heater-cooler units (HCUs) used cardiac heart surgery was traced as source. The aim was to investigate occurrence of M. chimaera and other microorganisms in HCUs and evaluate the silver-ion cleaning routine.Five HCUs were disinfected with silver-ions and examined for mycobacteria directly (15 min) after the disinfection procedures and later on three occasions (3, 6, 10 weeks). One HCU was selected for additional investigation of the presence of other microorganisms. In addition, tap water from five sinks in the surgical department was examined for the presence of mycobacteria and other microorganisms.M. chimaera grew in all the HCU water tanks and in 35 of the 40 HCU samples. Three of the samples also contained Mycobacterium gordonae. When the selected HCU tanks were analysed directly after the disinfection procedure bacteria and fungi were found but no non-fermenting Gram-negative rods. These HCU samples contained a doubled to 3 fold amount of bacteria compared to initial tap water samples. No mycobacteria were found in any sample from the five water taps.The silver-ion cleaning routine was insufficient and M. chimaera was found in all HCUs. However, no mycobacteria were found in any sample from the five water taps suggesting another source of colonization. It is probable that residual water and biofilm are of importance. Our results emphasize the need for improved disinfection procedures and improved construction of the HCUs.

Background Nontuberculous Mycobacteria (NTM) are environmental microorganisms that can affect human health. A 2009–2010 occurrence survey of NTM in potable tap water samples indicated an increased recovery rate for many clinically significant species such as M. avium (30%) and M. abscessus (12%). To determine if these trends by species were mirrored in human infections, isolation rates of NTM species identified in clinical laboratory reports from four states were evaluated. Method Clinical laboratory reports from the Mississippi, Missouri, Ohio, and Wisconsin Health Departments were used to investigate the species of NTM isolated from human specimens in 2014. The NTM positive specimen reports were tabulated for each species and complex/group. The number of reports by month were used to investigate seasonal trends. The 2014 isolation rates were compared to historic values to examine longitudinal trends. Results The positive rate of NTM specimens increased from 8.2 per 100,000 persons in 1994 to 16 per 100,000 persons in 2014 (or 13.3 per 100,000 after excluding Mycobacterium gordonae). Changes in NTM diversity were observed in complex/groups known to be clinically significant. Between 1994 and 2014 the rate implicating M. abscesses-chelonae group and M. avium complex increased by 322 and 149%, respectively. Conclusions Based on public health data supplied by the four State’s Health Departments and the 2014 U.S. population, 50,976 positive NTM specimen reports per year were projected for the nation; serving as an indicator for the national potential disease burden that year. Electronic supplementary material The online version of this article (10.1186/s12879-018-3043-7) contains supplementary material, which is available to authorized users.

Background: There is evidence that drinking water could be a source of infections with pathogenic nontuberculous mycobacteria (NTM) potentially risky to human health. The aim was to investigate the resistance of two NTM isolated from drinking water, Mycobacterium gordonae and Mycobacterium chubuense, at different concentrations of chlorine (as sodium hypochlorite), used in drinking water sanitation. Methods: The NTM were grown in suspension and in biofilms and were challenged with biocide for 10 and 60 min. Results: To obtain 7-log reduction from the initial population of M. chubuense, in the planktonic state, there were necessary 20 ppm of chorine and 60 min of exposure. The same effect was achieved in M. gordonae with 10 ppm for the same period. The maximum reduction of both NTM in biofilm was 3-log reduction and was achieved using 30 ppm for 60 min. The chlorine susceptibility of cells in biofilms was significantly lower than that of planktonic cells. The results highlight the resistance of both NTM to the concentrations used in routine water sanitation (0.2 ppm according to Argentine Food Code). Differences in chlorine resistance found between the two NTM in planktonic growth decrease when they are grown in biofilm. Conclusion: This suggests that current water disinfection procedures do not always achieve effective control of NTM in the public supply system, with the consequent health risk to susceptible population, and the need to take into account biofilms, because of their deep consequences in the way to analyze the survival of prokaryotic cells in different environments.

We evaluated PyroMark Q24 (QIAGEN) pyrosequencing as a method for the identification of mycobacteria, with potential application in clinical practice. Sequence data from the hypervariable region A of the 16S rRNA gene (43 and 35bp sequences) were obtained using PyroMark Q24, and a similarity search was performed automatically with PyroMark IdentiFire software. Of the 148 mycobacterial type strains tested, 138 (93.2%) were accurately identified to single or clade species level, including complex level. From the remaining 10 strains, 3 (Mycobacterium gilvum, Mycobacterium goodi, and Mycobacterium thermoresistible) showed poor sequencing quality of homopolymers. For 6 other strains (Mycobacterium cosmeticum, Mycobacterium flavescens, Mycobacterium pallens, Mycobacterium hodleri, Mycobacterium xenopi, and Mycobacterium crocinum), the sequences were unreadable from the middle, and Sanger sequencing indicated biallelic site. Finally, a 40bp sequence for Mycobacterium gordonae could not be obtained despite repeated attempts. PyroMark Q24 provided accurate identification of multiple mycobacterial strains isolated from common clinical settings, but additional gene sequencing is required to distinguish species identified as a group or complex.

TOPIC: Chest Infections TYPE: Fellow Case Reports INTRODUCTION: Mycobacterium gordonae, a commonly found species of mycobacteria. While frequently encountered in the environment and clinical laboratories it is almost always considered nonpathogenic. Despite being a non-virulent organism, there are many cases of clinically significant disease. Occasionally, it has been known to cause severe infections, especially in patients with an underlying predisposition or immunosuppression such as AIDS. CASE PRESENTATION: A 39-year-old male with no known PMH presents to the hospital with complaint of worsening dyspnea, fever, and nonproductive cough. He was in his usual state of health until 3 weeks prior when he began to experience body aches, sore throat, fever and shortness of breath. Continued symptoms prompted visit to the hospital at which time he discharged from the emergency department. The patient continued to have progressive worsening of dyspnea with fever prompting return to the hospital and subsequent admission for acute hypoxic respiratory failure. He lives at home with his wife and 4 children who do not have similar symptoms. He has lived in the US for 17 years and denies recent travel, sick contacts, history of autoimmune diseases, smoking history, vaping history, drug use, skin rash, joint pain. On admission vital signs and physical exam showed tachycardia and tachypnea with fever and inspiratory rales. Laboratory data including comprehensive metabolic panel and complete blood count with differential was unrevealing. Chest imaging showed diffuse bilateral dense ground glass opacities. Inflammatory markers including D-dimer, Ferritin, LDH, CRP were elevated. Influenza, COVID-19, mycoplasma, legionella antigen resulted negative. Testing for HIV returned positive with a CD4 count of 8. Given the clinical picture the patient was started on steroids, Bactrim, and Highly active antiretroviral therapy (HAART). Bronchoscopy with BAL was performed showing negative cytology, negative GMS-silver stain. However, AFB testing showed positive M. gordonae with fungal culture showing Candida albicans. Prednisone was discontinued and the patient was continued on HAART therapy along with Bactrim and sent for further treatment with Infectious Disease specialists. DISCUSSION: While there are reports of M. gordonae infections involving many organ systems, pulmonary infection is the most common site of symptomatic disease. Common symptoms include cough, weight loss, dyspnea, hemoptysis and fever. In addition to clinical symptoms and radiographic abnormalities, positive cultures from sputum, bronchial wash, transbronchial or lung biopsy are necessary for diagnosis. CONCLUSIONS: M.gordonae is capable of causing significant infections in both immunocompromised and immunocompetent hosts. Although the likelihood of this being a contamination is significant, its isolation should prompt further evaluation especially in those patients who are immunocompromised. REFERENCE #1: Weinberger M, Berg SL, Feurstein IM, Pizzo PA, Witebsky FG. Disseminated infection with Mycobacterium gordonae: Report of a case and critical review of the literature. Clin Infect Dis. 1992;14:1229–1239. REFERENCE #2: Asija A, Prasad A, Eskridge E. Disseminated Mycobacterium gordonae infection in an immunocompetent host. Am J Ther. 2011 May. REFERENCE #3: Brener ZZ, Zhuravenko I, Bergman M. Acute kidney injury in a patient with nontuberculous mycobacterial infections: a case report. Cases J. 2009 Jan 23. DISCLOSURES: No relevant relationships by mohammed halabiya, source=Web Response No relevant relationships by Richard Miller, source=Web Response No relevant relationships by Shawn Pate, source=Web Response

Nontuberculous mycobacteria (NTM) have emerged as an important cause of opportunistic nosocomial infections. NTM has frequently been isolated from hospital water distribution systems. The aim of this study was to survey the risk of NTM infections and determine the prevalence of NTM species in the hospital water distribution systems in Tabriz, Iran. One hundred and twenty samples of water from different sources of Tabriz hospitals were collected. The samples were filtered through 0.45-µm pore size membranes and decontaminated with 0.01% cetylpyridinium chloride. The sediment was inoculated onto Lowenstein–Jensen medium and incubated for 8 weeks. For identification to the species level, partial sequence analysis of the hsp65 and 16S rRNA genes were used. NTM were detected in 76 (63.3%) of 120 samples. Potentially pathogenic mycobacteria and saprophytic mycobacteria were isolated. Mycobacterium gordonae was the only single species that was present in all types of water. The prevalence of NTM in Tabriz hospitals' water compared with many investigations on hospital waters was high. This indicates that the immunocompromised patients and transplant recipients are at risk of contamination which necessitates considering decontamination of water sources to prevent such potential hazards.

Limited data are available describing extrapulmonary nontuberculous mycobacteria (NTM) infections in the general population. We describe results from statewide population-based laboratory surveillance in Oregon, USA, during 2007–2012. We defined a case of extrapulmonary NTM infection as >1 isolate from skin/soft tissue, disseminated sites, lymph node, joint, or other sites. The annual incidence of extrapulmonary NTM infection (other than Mycobacterium gordonae) was stable, averaging 1.5 cases/100,000 population. Median age of the 334 patients was 51 years, and 53% of patients were female. Half of cases were caused by M. avium complex, but rapid-growing NTM species accounted for one third of cases. Most extrapulmonary NTM infections are skin/soft tissue. Compared with pulmonary NTM infection, more extrapulmonary infections are caused by rapid-growing NTM species. the designation of NTM as a reportable disease in Oregon in 2014 will result in better detection of changes in the incidence and patterns of disease in the future.

The redclaw crayfish Cherax quadricarinatus (von Martens, 1868) is one of the most commonly exploited freshwater crayfish species worldwide. Redclaw crayfish are susceptible to a number of pathogens but none have been linked to widespread epizootics. Mycobacterial infections have been sporadically reported in crayfish. In the case described, histopathology and bacterial identification confirmed an opportunistic infection caused by Mycobacterium gordonae in a hatchery of C. quadricarinatus in Israel. Intranuclear inclusion bodies, recorded in cells of the tubular epithelium of the hepatopancreas by histopathology, indicate a co-infection with a viral agent, referable to C. quadricarinatus bacilliform virus (CqBV). To the best of our knowledge this is the first description of mycobacteriosis in redclaw crayfish.

Background: There is evidence that tap water is the vehicle through which nontuberculous mycobacteria (NTM) infect or colonize the human body. The objective of this study was to determine the presence and diversity of NTM in the water distribution system of Bahía Blanca city, Argentina (sites S2/S3) and in the dike that supplies water to it (S1). Methods: Culture-dependent method, biochemical tests, and molecular method (16S rRNA sequencing gene) were combined to detect and identify NTM. Results: NTM were isolated in 51.6% (64/124) of all the samples analyzed. Mycobacterium gordonae was the most frequently isolated organism (15/64) in all samples analyzed, followed by Mycobacterium peregrinum and Mycobacterium frederiksbergense. Significant differences were found in the residual chlorine values between sampling S2 and S3. In both sites, maximum counts were recorded but they did not correlate with low chlorine values. A concentration higher than 500 colony-forming unit/L of NTM was never found, which can be attributed to the negative effect caused by decontamination methods being a point to consider for the recovery of NTM. In 46.9% (30/64) of samples, both methods coincided in the identification, and the obtained sequences presented ≥99% identity. Identification at the species level was achieved in 50% (32/64) of the isolates. Nearly 17.2% (11/64) of the isolates showed a similarity

Background Non-tuberculous mycobacterial pulmonary disease (NTM PD) presents a difficult clinical challenge. The presentation varies from transient respiratory pathogen to aggressive destruction, relying heavily on clinical symptoms, effective sputum culture and imaging. Although CT is a reliable and non-invasive method for diagnosing non-CF bronchiectasis, features of NTM PD on CT imaging are nonspecific and often undistinguishable from underlying respiratory conditions, concurrent infection or inflammation. NTM prevalence is increasing worldwide and non- CF bronchiectasis remains an important risk factor. Despite improved BTS/ATS guidance for treatment, the lack of specific CT findings in non-CF bronchiectasis patients makes decision to treat problematic. Patients and methods A retrospective analysis of CT reports of 261 consecutive non-CF bronchiectasis patients at Royal United Hospital, Bath was undertaken. Reports were scrutinised for the presence of seven CT characteristics: tree in bud, pulmonary nodules or nodularity, consolidation, cavitation, ground glass change, pleural thickening and pleural effusion. NTM culture status was recorded for each patient but Mycobacterium gordonae cases were classified as non-NTM. CT findings were compared between NTM positive sputum culture cases and NTM negative cases using Chi-squared for statistical significance between groups (SPSS version 24). Results 261 patients with non-CF bronchiectasis were identified. Of these 27 (10.3%) had positive NTM sputum culture (MAC 16 (59.3%), M. abscessus 5 (18.5%), M. chelonae 4 (14.8%), M. malmoense 1 (3.7%), M. xenopi 1 (3.7%)). In total the incidence of specific CT findings was as follows; tree and bud 50 (19.2%), nodules/nodularity 97 (37.2%), consolidation 51 (19.5%), cavitation 8 (3.1%), ground glass 30 (11.5%), pleural thickening 13 (5.0%), pleural effusion 10 (3.8%). 103 patients (39.5%) had none of these characteristics reported. There was no statistically significant difference in the presence or absence of CT signs in patients with NTM and no NTM (see table 1). Conclusions In non-CF bronchiectasis there appears to be no reliable CT findings to identify patients with NTM infection. There a remains a need for a high level of clinical suspicion in patients with bronchiectasis and microbiology remains the mainstay of diagnosis.

Diagnosis and identification of mycobacterial infections is often a challenge that engages different medical fields. Due to the time-consuming diagnostic procedures for most mycobacteria and the lack of rapid differentiation between Mycobacterium tuberculosis and NTMs species, incorrect diagnosis of non-tuberculosis leads to the use of TB treatment protocol and prolonged hospitalization. So, the aim of this study was to rapid identification of Mycobacterium tuberculosis strains from non-Mycobacterial tuberculosis by the multiplex Real time PCR method to prevent misdiagnosis in the treatment procedures. In this study, 82 sputum specimens were collected from patients suspected to tuberculosis referred to Peymanieh Hospital and health centers of Jahrom, as well as those treated with TB drugs referred to Jahrom Tuberculosis Center in 2015. Sediment was collected from the clinical samples by Petroff's method, DNA extraction was done with commercial DNA kit. 10 pairs primer; 23srRNA, grvB, ITS 152 bp, ITS172bp 164 bp, ITS, dnaJ, katG, rpoB, ITS 185 bp, ITS 185 bp were used. Multiplex real time PCR was performed in one run and in three different reactions and their melting curve was evaluated for both species and drug resistance. Among 82 suspected tuberculosis patients referred to Jahrom Tuberculosis Center, 26 (31.7% of total samples) were Mycobacterium positive, of which 13 (15.85%) were Mycobacterium tuberculosis, 7 (53.58%) Mycobacterium kansasii, 1 specimen (1.21%) were Mycobacterium gordonae and 5 specimens (6.09%) were uncommon Mycobacterium tuberculosis. No MDR was found in Mycobacterium tuberculosis specimens and no non-tuberculosis resistant specimens were found in Mycobacterium tuberculosis specimens. According to the results of this study, the prevalence of Mycobacterium tuberculosis and non-tuberculosis in Jahrom city was different from other parts of the country. Also, this procedure is convenient, easy and quick to detect common mycobacterial strains to prevent ectopic treatments and is also a good way to follow up treatment in treated cases.

The whole blood model for infection has proven useful to analyze the immunological response to Mycobacterium tuberculosis, because it exerts a significant antimicrobial activity. Although this activity has been generally assumed to be cellular, we have found that the leukocyte fraction of blood from healthy volunteers did not kill the bacilli. We have discovered that plasma was responsible for a large proportion, but not all, of the antimicrobial activity. Furthermore, infected monocytes controlled the mycobacterial multiplication when cultivated in the presence of plasma. Intriguingly, serum from the same donors did not share this activity, although it was able to eliminate the non-pathogenic Mycobacterium gordonae To identify the remaining components that participate in the antimycobacterial activity we fractionated blood in leukocytes, plasma, erythrocytes and platelets, and analyzed the bactericidal power of each fraction and their combinations using a factorial design. We found that erythrocytes, but not platelets, participated and showed by flow cytometry that mycobacteria physically associated with erythrocytes. We propose that in exposed healthy individuals that show 'early clearance' of the mycobacteria, the innate response is predominantly humoral, probably through the effect of antimicrobial peptides and proteins.

Non-tuberculous mycobacteria (NTM) are important pathogens in lung transplant recipients. This study describes the spectrum of NTM respiratory tract infections and examines the association of NTM infections with lung transplant complications.Data from 208 recipients transplanted from November 1990 to November 2005 were analyzed. Follow-up data were available to November 2010. Lung infection was defined by bronchoalveolar lavage, sputum, or blood cultures in the appropriate clinical setting. All identified NTM respiratory tract infections were tabulated. The cohort of patients with NTM lung infections (NTM+) were compared to the cohort without infection (NTM-). Univariate and multivariate analysis was performed to determine characteristics associated with NTM infection. Survival analyses for overall survival and development of bronchiolitis obliterans syndrome (BOS) were also performed.In total, 52 isolates of NTM lung infection were identified in 30 patients. The isolates included Mycobacterium abscessus (46%), Mycobacterium avium complex (MAC) (36%), Mycobacterium gordonae (9%), Mycobacterium chelonae (7%), and Mycobacterium fortuitum (2%), with multiple NTM isolates seen on 3 different occasions. The overall incidence was 14%, whereas cumulative incidences at 1, 3, and 5 years after lung transplantation were 11%, 15%, and 20%, respectively. Comparisons between the NTM+ and NTM- cohorts revealed that NTM+ patients were more likely to be African-American and have cytomegalovirus mismatch. Although no difference was seen in survival, the NTM+ cohort was more likely to develop BOS (80% vs. 58%, P = 0.02). NTM+ infection, however, was not independently associated with development of BOS by multivariate analysis.With nearly 20 years of follow-up, 14% of lung recipients develop NTM respiratory tract infections, with M. abscessus and MAC more commonly identified. M. gordonae was considered responsible for nearly 10% of NTM infections. Although survival of patients with NTM infections is similar, a striking difference in BOS rates is present in the NTM+ and NTM- groups.

Summary Background Nontuberculous mycobacteria (NTM) are environmental mycobacteria associated with a range of infections. Reports of NTM epidemiology have primarily focused on pulmonary infections and isolations, however extrapulmonary infections of the skin, soft tissues and sterile sites are less frequently described. Methods We comprehensively reviewed laboratory reports of NTM isolation from North Carolina residents of three counties during 2006–2010. We describe age, gender, and race of patients, and anatomic site of isolation for NTM species. Results Among 1033 patients, overall NTM isolation prevalence was 15.9/100,000 persons (13.7/100,000 excluding Mycobacterium gordonae ). Prevalence was similar between genders and increased significantly with age. Extrapulmonary isolations among middle-aged black males and pulmonary isolations among elderly white females were most frequently detected. Most isolations from pulmonary sites and blood cultures were Mycobacterium avium complex; rapidly growing NTM (e.g. Mycobacterium chelonae , Mycobacterium fortuitum ) were most often isolated from paranasal sinuses, wounds and skin. Conclusions We provide the first characterization of NTM isolation prevalence in the Southeastern United States (U.S.). Variation in isolation prevalence among counties and races likely represent differences in detection, demographics and risk factors. Further characterization of NTM epidemiology is increasingly important as percentages of immunocompromised individuals and the elderly increase in the U.S. population.

The Epistem Genedrive assay rapidly detects the Mycobacterium tuberculosis complex from sputum and is currently available for clinical use. However, the analytical and clinical performance of this test has not been fully evaluated. The analytical limit of detection (LOD) of the Genedrive PCR amplification was tested with genomic DNA; the performance of the complete (sample processing plus amplification) system was tested by spiking M. tuberculosis mc 2 6030 cells into distilled water and M. tuberculosis -negative sputum. Specificity was tested using common respiratory pathogens and nontuberculosis mycobacteria. A clinical evaluation enrolled adults with suspected pulmonary tuberculosis, obtained three sputum samples from each participant, and compared the accuracy of the Genedrive to that of the Xpert MTB/RIF assay using M. tuberculosis cultures as the reference standard. The Genedrive assay had an LOD of 1 pg/μl (100 genomic DNA copies/reaction). The LODs of the system were 2.5 × 10 4 CFU/ml and 2.5 × 10 5 CFU/ml for cells spiked into water and sputum, respectively. False-positive rpoB probe signals were observed in 3/32 (9.4%) of the negative controls and also in few samples containing Mycobacterium abscessus , Mycobacterium gordonae , or Mycobacterium thermoresistibile . In the clinical study, among 336 analyzed participants, the overall sensitivities for the tuberculosis case detection of Genedrive, Xpert, and smear microscopy were 45.4% (95% confidence interval [CI], 35.2% to 55.8%), 91.8% (95% CI, 84.4% to 96.4%), and 77.3% (95% CI, 67.7% to 85.2%), respectively. The sensitivities of Genedrive and Xpert for the detection of smear-microscopy-negative tuberculosis were 0% (95% CI, 0% to 15.4%) and 68.2% (95% CI, 45.1% to 86.1%), respectively. The Genedrive assay did not meet performance standards recommended by the World Health Organization for a smear microscopy replacement tuberculosis test. Epistem is working on modifications to improve the assay.

Mycobacterium gordonae is often regarded as a weak pathogen that only occasionally causes overt disease. We report a case of M. gordonae infection in the facial skin, nasal mucosa, and paranasal sinus in an immunocompetent patient and review previous cases. The T-SPOT.TB test might be useful in diagnosing such cases.

Non-tuberculous mycobacteria (NTM) are increasingly a cause of human respiratory tract colonization and mycobacterial lung disease (NTM-LD), especially in patients with chronic lung diseases. The aim of the present study was to find the factors predictive of NTM-LD in patients with obstructive lung diseases and NTM respiratory isolates. A total of 839 isolates of NTM, obtained from 161 patients between 2010 and 2020 in a single pulmonary unit, have been retrospectively reviewed. Of these isolates, 73 concerned 36 patients with obstructive lung diseases (COPD-26, asthma-3, COPD/asthma overlap syndrome-7). NTM-LD was recognized according to the American Thoracic Society (ATS) and the Infectious Diseases Society of America (IDSA) criteria in 17 patients, colonization in 19. Lower BMI, elevated body temperature on admission, infiltrative/cavitary lesions on chest CT, and NTM species other than Mycobacterium gordonae were the significant predictors of NTM-LD recognition. Based on the above-mentioned predictive factors, an original scoring system was implemented. The diagnostic utility of the scoring system was higher than that of single parameters. We conclude that NTM-LD prediction in patients with obstructive lung diseases and positive respiratory isolates is difficult. A scoring system based on clinical, radiological and microbiological characteristics was capable of facilitating the differential diagnosis, but it needs further validation in a larger study group.

Over the last 30 years, there have been at least 17 published reports of nontuberculous mycobacteria (NTMs) being isolated from hospital ice or ice-making machines. Of these, 12 were reports of pseudo-outbreaks, i.e., the nosocomial transmission of organism from hospital ice/ice machines to patients, resulting in patient colonization, but with no disease manifestations. In addition, there were five outbreaks that resulted in clinical disease/pathology associated with NTM organism. Eleven different species of NTMs have been associated with these reports, where over half (59%) of the species identified were Mycobacterium fortuitum (18%), Mycobacterium gordonae (14%), Mycobacterium mucogenicum (14%), and Mycobacterium porcinum (14%). Several of these reports clearly documented that ice machines had been properly maintained, cleaned, and serviced in accordance with the CDC guidelines yet became contaminated with NTM organisms. These reports frequently detail that after extensive cleaning/disinfection following the discovery of NTM organisms, ice machines remained contaminated with NTM organisms, highlighting the difficulty in eradicating these from ice machines, once contaminated. Several reports identified that the only remedy to the contamination problem was to replace the ice machine with a new machine. Two qualitative risk assessment models are presented for (i) patients exposed to contaminated ice machine but before NTM colonization/infection and (ii) patients already colonized with NTMs from ice machines. Therefore, to protect immunocompromised/immunosuppressed patients' safety, especially during surgical or respiratory procedures, ice should not be sourced from the ice machine but should be made from sterile water and stored safely and separately away from the ice machine.

Background: Mycobacterium spp. are responsible for several diseases, particularly in immunocompromised populations. The spreading of the resistance to antimycobacterial drugs is a significant problem to the public health and requires to find out a new and innovative alternative for the treatment of drug resistant mycobacterial strains. In this study, the antimycobacterial activity of Micromeria barbata, Eucalyptus globulus and Juniperus excelsa essential oils extracted from Lebanese plants was investigated against selected Mycobacterium spp. strains. Methods: Several dilutions of the three aforementioned essential oils were studied for antimycobacterial activity against four Mycobacterium spp. strains: Mycobacterium tuberculosis subsp. tuberculosis (ATCC® 27294™), multidrug-resistant M. tuberculosis (CMUL 157), Mycobacterium kansasii Hauduroy (ATCC® 12478™) and Mycobacterium gordonae Bojalil et al. (ATCC® 14470™). Results: Even with high dilutions, all tested essential oils showed a high antimycobacterial activity against targeted strains. Our data showed that M. barbata, E. globulus and J. excelsa essential oils totally inhibit the mycobacterial growth whatever the tested strains for the dilution 1/250, 1/100 and 1/250, respectively. Conclusion: To our knowledge, this is the first study regarding the antimycobacterial activity of essential oils in Lebanon. Our data show promising results, and encourage to investigate more on these medicinal plants, especially M. barbata. Keywords: Essential oils, Antimycobacterial activity, Micromeria barbata, Eucalyptus globulus, Juniperus excelsa

Background Non-tuberculous mycobacteria (NTM) infection is an increasing problem worldwide. The epidemiology of NTM in most Asian countries is unknown. This study investigated the epidemiology, and clinical profile of inpatients in whom NTM was isolated from various anatomical sites in a Singaporean population attending a major tertiary referral centre. Methods Demographic profile, clinical data, and characteristics of patients hospitalized with NTM isolates at a major tertiary hospital over two-year period were prospectively assessed (2011–2012). Data collected included patient demographics, ethnicity, smoking status, co-morbidities, NTM species, intensive care unit (ICU) treatment, and mortality. Results A total of 485 patients (62.1% male) with 560 hospital admissions were analysed. The median patient age was 70 years. Thirteen different NTM species were isolated from this cohort. Mycobacterium abscessus (M. abscessus) (38.4%) was most frequently isolated followed by Mycobacterium fortuitum (M. fortuitum) (16.6%), Mycobacterium avium complex (MAC) (16.3%), Mycobacterium kansasii (M. kansasii) (15.4%), and Mycobacterium gordonae (M. gordonae) (6.8%). Most (91%) NTM was isolated from the respiratory tract. The three most common non-pulmonary sites were; blood (2.7%), skin wounds and abscesses (2.1%), and gastric aspirates (1.1%). A third (34.4%) of the study population had prior pulmonary tuberculosis (PTB). There was a significant association between isolated NTM species, and patient age (p=0.0002). Eleven (2.2%) patients received intensive care unit (ICU) treatment during the study period and all cause mortality within 1 year of the study was 16.9% (n=82). Of these, 72 (87.8%) patients died of pulmonary causes. Conclusions The profile of NTM species in Singapore is unique. M. abscessus is the commonest NTM isolated, with a higher prevalence in males, and in the elderly. High NTM prevalence is associated with high rates of prior PTB in our cohort. Published version

Mycobacterium avium complex-related diseases are often associated with poorly maintained hot water systems. This calls for the development of new control strategies. The aim of this study was to investigate the activity of essential oils (EOs) from the Mediterranean plants, common juniper, immortelle, sage, lavandin, laurel, and white cedar against Mycobacterium avium ssp. avium, Mycobacterium intracellulare, and Mycobacterium gordonae in culturing broth and freshwater as their most common habitat. To do that, we developed a new method of water microdilution to determine their minimal effective concentrations (MEC). The most active EO was the one from the common juniper with the MEC of 1.6 mg mL-1. Gas chromatography / mass spectrometry the juniper EO identified monoterpenes (70.54 %) and sesquiterpenes (25.9 %) as dominant component groups. The main monoterpene hydrocarbons were α-pinene, sabinene, and β-pinene. The juniper EO significantly reduced the cell viability of M. intracellulare and M. gordonae at MEC, and of M. avium at 2xMEC. Microscopic analysis confirmed its inhibitory effect by revealing significant morphological changes in the cell membrane and cytoplasm of all three bacteria. The mode of action of the juniper EO on the cell membrane was confirmed by a marked leakage of intracellular material. Juniper EO has a great practical potential as a complementary or alternative water disinfectant in hot water systems such as baths, swimming pools, spa pools, hot tubs, or even foot baths/whirlpools., Ispitali smo aktivnost eteričnih ulja borovice [Juniperus communis (J. communis)], smilja (Helichrysum italicum), kadulje (Salvia officinalis), lavandina (Lavandula hybrida), lovora (Laurus nobilis) i tuje (Thuja occidentalis) prema sojevima Mycobacterium avium ssp. avium (M. avium), Mycobacterium intracellulare (M. intracellulare) i Mycobacterium gordonae (M. gordonae). Za određivanje minimalnih inhibicijskih koncentracija (MIK) i minimalnih baktericidnih koncentracija (MBK) testiranih eteričnih ulja koristili smo se metodom mikrodiluacije u tekućem bujonu (Middlebrook 7H9), a za određivanje minimalne efektivne koncentracije (MEK) umjesto bujona korištena je sterilna voda iz slavine kojoj je dodano 0,05 % Tweena 80. Dvostruka serijska razrjeđenja eteričnih ulja, počevši od 0,1 do 51,2 mg mL-1, u tekućem bujonu ili u sterilnoj vodi iz slavine inokulirana su s mikobakterijskom suspenzijom uz dodatak resazurina. Najučinkovitije eterično ulje prema svim ispitivanim sojevima mikobakterija bilo je eterično ulje J. communis s MBK / MIK / MEK vrijednošću od 1,6 mg mL-1. Metodom plinske kromatografije i masene spektrometrije analizirano je eterično ulje J. communis. Udio monoterpena iznosio je 70,54 %, a seskviterpena 25,9 %. Glavni monoterpenski ugljikovodici bili su α-pinen, sabinen i β-pinen. Eterično ulje J. communis pokazalo je značajan inhibicijski učinak na M. intracellulare i M. gordonae pri MEK i na M. avium pri dvostrukim MEK. Elektronskom mikroskopijom kod svih triju sojeva mikobakterija nakon izlaganja djelovanju eteričnog ulja J. communis otkrivene su značajne morfološke promjene stanične membrane i citoplazme. Učinak eteričnog ulja J. communis na destrukciju stanične membrane mikobakterija potvrđen je značajnim otpuštanjem unutarstaničnog materijala mjerenjem apsorbancije supernatanta pri 260 nm i 280 nm. Zaključno, u našem smo radu razvili novu metodu za ispitivanje antimikrobnog učinka eteričnih ulja ili drugih prirodnih tvari na netuberkulozne mikobakterije koja oponaša uvjete kao u vodenim sustavima. Prirodni proizvodi, osobito eterična ulja, imaju ne samo velik potencijal kao antimikrobni agensi nego i moguću praktičnu primjenu kao alternativni dezinficijensi.

Case: A 43-year-old man with a history of well-controlled HIV (human immunodeficiency virus) infection presented with sacroiliac joint destruction from a Mycobacterium gordonae infection. A sacroiliac joint arthrodesis was performed using a minimally invasive technique utilizing both biologic fusion (allograft bone with rhBMP-2 [recombinant human bone morphogenetic protein-2]) and fixation with titanium ingrowth rods. Conclusion: To our knowledge, this is the first reported case of infectious sacroiliitis from a nontubercular mycobacterium (M. gordonae) treated with a combination of joint debridement, biologic fusion with bone graft, and nonbiologic functional fusion using titanium ingrowth rods, all performed in a minimally invasive fashion. This strategy effectively alleviated pain and preserved function at 2 years of follow-up.

TNF-α는 항산균에 대한 인체의 면역학적 방어기전에서 중요한 역할을 한다. Mycobacterium gordonae는 독성이 낮아임상 검체에서 검출되면 대부분 오염균으로 간주되고 있다. 저자들은 TNF-α 길항제로 치료 중인 궤양성 대장염 환자에서 M. gordonae 폐질환으로 진단되어 적절한 약물 치료 후-호전된 증례를 경험하였기에 문헌고찰과 함께 보고하는 바이다.

The first total synthesis of the branched oligosaccharide OSE-1 of Mycobacterium gordonae (strain 990) is reported. An intramolecular aglycon delivery approach was used for constructing the desymmetrized 1,1'-α,α-linked trehalose moiety. A [3+2] glycosylation of the trisaccharide donor and trehalose acceptor furnished the right hand side pentasaccharide. Regioselective O3 glycosylation of L-rhamnosyl 2,3-diol allowed expedient synthesis of the left hand side tetrasaccharide. The nonasaccharide was assembled in a highly convergent fashion through a [4+5] glycosylation.

Objectives: Detection of the humoral response to diagnose active tuberculosis has had varied success. We sought to further characterize the performance of a commercial serologic assay (Active TBDetect IgG ELISA; InBios International, Seattle, WA), which had demonstrated promising results in prior studies. Methods: Blood specimens from patients with mycobacterial infections, autoimmune disorders, and documented nonmycobacterial infections were prospectively collected for testing by the Active TBDetect IgG ELISA. Pertinent medical records were reviewed. Results: The sensitivity of the InBios IgG ELISA for active tuberculosis cases was 54.1% (20/37). Reactivity occurred in 24.1% (14/58) of nontuberculous mycobacterium cases, 10.4% (7/67) of nonmycobacterial infections, 10.5% (11/105) of autoimmune disorder cases, 8.7% (8/92) of noninfected patients, 14.3% (1/7) of patients with latent tuberculosis, and 10.7% (3/28) of control pediatric cases. Overall specificity was 87.5% (288/329). Receiver operator curve analysis demonstrated an area under the curve of 0.74. Reactivity with nontuberculous mycobacterium infection occurred with Mycobacterium avium-intracellulare complex, Mycobacterium chelonae/abscessus complex, Mycobacterium simiae, and Mycobacterium gordonae and was positively associated with having a positive acid-fast bacilli smear. Conclusions: This study confirmed the limitations of serodiagnosis for active tuberculosis, including poor sensitivity and increased reactivity with nontuberculous mycobacterium-positive patients.

Mycobacteria are associated with a number of well-characterized diseases, yet we know little about their stress biology in natural ecosystems. This study focuses on the isolation and characterization of strains from Yellowstone National Park (YNP) and Glacier National Park (GNP; USA), the majority of those identified were Mycobacterium parascrofulaceum, Mycobacterium avium (YNP) or Mycobacterium gordonae (GNP). Generally, their windows for growth spanned a temperature range of > 60 °C; selected isolates grew at super-saturated concentrations of hydrophobic stressors and at levels of osmotic stress and chaotropic activity (up to 13.4 kJ kg(-1) ) similar to, or exceeding, those for the xerophilic fungus Aspergillus wentii and solvent-tolerant bacterium Pseudomonas putida. For example, mycobacteria grew down to 0.800 water activity indicating that they are, with the sole exception of halophiles, more xerotolerant than other bacteria (or any Archaea). Furthermore, the fatty-acid composition of Mycobacterium cells grown over a range of salt concentrations changed less than that of other bacteria, indicating a high level of resilience, regardless of the stress load. Cells of M. parascrofulaceum, M. smegmatis and M. avium resisted the acute, potentially lethal challenges from extremes of pH ( 13), and saturated MgCl2 solutions (5 M; 212 kJ kg(-1) chaotropicity). Collectively, these findings challenge the paradigm that bacteria have solute tolerances inferior to those of eukaryotes.

BACKGROUND Although Mycobacterium gordonae isolation from respiratory samples is usually regarded as contamination, M. gordonae can cause definite pulmonary disease. The establishment of a standard diagnostic criteria of pulmonary disease that is caused by this low virulence mycobacterium is obviously necessary. METHODS We performed clinical research on over 200 cases in which M. gordonae was isolated over an 8-year period, focusing on the M. gordonae subtype. Sequence analysis of rpoB was performed to identify the genotypes. RESULTS A total of 287 respiratory samples (209 cases) were positive for M. gordonae. Twenty-seven cases (12.9%) had a positive culture more than twice and 11 of these cases (5.3%) had more than three positive cultures. Ultimately, three cases (1.4%) were newly diagnosed as M. gordonae pulmonary disease using our own diagnostic criteria. In all of the identified M. gordonae cases, the cultures tested positive with a Mycobacteria Growth Indicator Tube test at 24 days; however, in patients with definitive pulmonary disease, the cultures were positive at 9 days. A subtype analysis revealed that all of the definitive disease cases had subtype C. CONCLUSION The time taken to detect a positive culture and subtype of the isolates could be used as the diagnostic criteria for definite M. gordonae pulmonary disease.

Private residences in rural areas with water systems that are not adequately regulated, monitored, and updated could have drinking water that poses a health risk. To investigate water quality on the Crow Reservation in Montana, water and biofilm samples were collected from 57 public buildings and private residences served by either treated municipal or individual groundwater well systems. Bacteriological quality was assessed including detection of fecal coliform bacteria and heterotrophic plate count (HPC) as well as three potentially pathogenic bacterial genera, Mycobacterium, Legionella, and Helicobacter. All three target genera were detected in drinking water systems on the Crow Reservation. Species detected included the opportunistic and frank pathogens Mycobacterium avium, Mycobacterium gordonae, Mycobacterium flavescens, Legionella pneumophila, and Helicobacter pylori. Additionally, there was an association between HPC bacteria and the presence of Mycobacterium and Legionella but not the presence of Helicobacter. This research has shown that groundwater and municipal drinking water systems on the Crow Reservation can harbor potential bacterial pathogens.