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Evaluation of the immune response in mice infected with Mycobacterium marinum.

Foot pad infection of mice with Mycobacterium marinum carried out with a view to comparing the immune response on the humoral level of such mice, with that observed previously in mice infected with M. leprae, indicated that there was a similarity in terms of the first appearance and proliferation of immunocytes and the time at which the peak and decline in the antibody-producing cells occurred. The significant difference appeared to be in the immunoglobulin G response, which was absent in the M. leprae infected mice, but occurred simultaneously with the immunoglobulin M response at a high level, both during a primary and after a secondary challenge administered 15 days post-primary infection in the M. marinum infected mice. Further confirmation was obtained through additional studies on the specific immunoglobulin levels and determination of both immunoglobulin M and immunoglobulin G antibodies by hemagglutination. Although the growth temperature requirement of the two organisms and their ability to initiate infection in areas of the body with lower temperatures are similar, it is suggested that the type of infection induced by each one of these species in the mouse may disallow the serious consideration of the M. marinum infection model as a possible alternative experimental model for studying the role of host immunity to M. leprae infections in mice.

Animals

Mycobacterium marinum infections treated with tetracycline.

Five cases of Mycobacterium marinum skin infections were successfully treated with 2 gm of tetracycline hydrochloride daily for periods from four to 12 weeks. In each case, the isolated M marinum was sensitive in vitro to tetracycline at levels from 25 to 50 microgram/ml. Tetracycline is recommended as treatment for extensive or sporotrichoid M marinum infections.

4-Aminobenzoic Acid

[Sensivity of Mycobacterium kansaii and Mycobacterium marinum to different antituberculous drugs (author's transl)].

Mycobacteriosis includes clinical manifestations caused by especies of the genus Mycobacterium other than M. tuberculosis and M. bovis. Therapy for these conditions has not been clearly sistematized as it has for tuberculosis, particularly because of the natural resistance that the etiologic agents present to a large number of antituberculous drugs. The sensitivity of M. kansasii and M. marinum to eleven tuberculostatic agents was studied in order to determine which one were best suited for treatment of cases caused by these species. The drugs showing the strongest action against M. kansasii were rifampin, cycloserine, streptomycin, and prothionamide. M. marinum was even more sensitive except to isoniazid, which is ineffective. As a general rule, especially if the sensitivity of the isolated strain is unknown drugs with little or no action on M. kansasii and M. marinum should not be used. Particularly the use of isoniazid and PAS should be avoided.

Antitubercular Agents

Granulomatous synovitis: the role of atypical mycobacteria.

Clinical data on 25 patients with granulomatous synovitis and bursitis observed from 1970 through 1977 are reviewed. The lesions occurred about the extremities, the wrists and hands being involved most often. With three exceptions, the patients had no significant underlying disease. The lesions were chronic and often followed minor trauma. Many patients had had prior surgery, steriod injections, or both. At the time of surgery for the synovitis, the gross appearance proved to be relatively characteristic, and microscopic examination revealed granulomatous inflammation. Mycobacteria were found in 20 patients. In 15 of these cases, mycobacteria were isolated in cultures; in an additional five cases, acid-fast bacilli were seen in the excised tissue but cultures were negative. The species isolated were Mycobacterium tuberculosis (four cases), Mycobacterium kansaii (six), Mycobacterium marinum (one), Mycobacterium gordonae (one), Mycobacterium avium-intracellulare (two), and Mycobacterium chelonei (one). The clinical results after therapy that combined surgery and antimicrobial agents were generally favorable. It is stressed that surgical specimens should be sent for appropriate cultures as well as for histologic study.

Adolescent

Ultra-high field strength electroporation enables efficient DNA transformation and genome editing in nontuberculous mycobacteria.

Efficient DNA delivery is essential for genetic manipulation of mycobacteria and for dissecting their physiology, pathogenesis, and drug resistance. Although electroporation enables transformation efficiencies exceeding 10⁵ CFU per µg DNA in Mycobacterium smegmatis and Mycobacterium tuberculosis, it remains highly inefficient in many nontuberculous mycobacteria (NTM), including Mycobacterium abscessus. Here, we discovered that NTM such as M. abscessus exhibit exceptional tolerance to ultra-high electric field strengths and that hypertonic preconditioning partially protects cells from electroporation-induced damage. Using ultra-high electric field strength (3 kV/mm) electroporation, we achieved dramatic improvements in plasmid transformation efficiency-up to 106-fold in M. abscessus, 83-fold in Mycobacterium marinum, and 37-fold in Mycobacterium kansasii-compared to standard conditions (1.25  kV/mm). Transformation efficiency was further influenced by the choice of selectable marker. Ultra-high field strength electroporation also markedly enhanced allelic exchange in M. abscessus expressing Che9c RecET recombinases, increasing the recovery of gene deletion mutants by over 1,000-fold relative to conventional electroporation. In parallel, oligonucleotide-mediated recombineering for targeted point mutations produced nearly 10,000-fold more mutants under ultra-high field conditions. Together, these findings establish ultra-high field electroporation as a robust, broadly applicable platform for genetic engineering of NTMs. This method substantially enhances transformation efficiency and enables construction of advanced genetic tools-including expression libraries and CRISPRi knockdown libraries-in species that have historically resisted genetic manipulation.IMPORTANCEInfections caused by nontuberculous mycobacteria (NTM), including Mycobacterium abscessus, are increasing globally, yet genetic manipulation of these pathogens remains technically challenging due to inefficient DNA delivery and low gene editing success. The ultra-high electric field strength electroporation strategy described here overcomes these barriers, enabling dramatic improvements in both transformation and genome editing efficiency. This advance paves the way for high-throughput functional genomics in NTMs, including the construction of genome-wide knockout, CRISPRi knockdown, and expression libraries. Broad adoption of this approach will accelerate discovery of genetic determinants of virulence and drug resistance, facilitating the development of antimicrobials and vaccines.

Electroporation

[Cutaneous mycobacteriosis of aquaria. A disease transmitted from fish to man].

The Authors refer about one case of cutaneous infection from Mycobacterium marinum transmitted to man from tropical fishes of a fish tank. The Authors describe the lesions produced by mycobacterium in man and in fish, as well as the relevant hystological findings and the microbiological enquiries carried out for isolation and typing. The possibility of direct transmission of a tubercular infection from fish to man has been demonstrated. The Authors think it to be useful a careful sanitary control on the ichthyc fauna destined to domestic fish tanks.

Animals

Tropical fish tank granuloma.

A case of tropical fish tank granuloma due to mycobacterium marinum is presented. The source, epidemiology, diagnosis and treatment are discussed.

Animals

Minocycline therapy of aquarium granuloma. Case reports and literature review.

Two patients with Mycobacterium marinum infections from home tropical fish aquariums are presented herein. Both patients showed dramatic improvement with minocycline therapy. In vitro susceptibility results also confirmed minocycline, doxycycline, and tetracycline efficacy using disc diffusion and dilution methods.

Adult

Immune response to persistent mycobacterial infection in mice.

Mycobacterium marinum has been recommended as a possible model of M. leprae for use in laboratory studies of antileprosy immunity. M. marinum introduced into the footpads of normal mice underwent a steady decline in viability, with less than 1% survival after a 30-day period. Small numbers of viable bacilli were recovered from the footpads of these mice up to 12 months later. Similarly, mice infected with M. simiae exhibited bacterial populations that persisted for up to 18 months with little change in viability. Injection of M. simiae into the footpads was followed by an extensive redistribution of the organisms in the tissues. Eventally, bacterial counts for footpads and draining lymph nodes stabilized, with small numbers of bacilli still present in the footpads 18 months later. Persistent growth, with little sign of any immune response, was also observed in mice infected with several strains of M. avium, as well as with one strain of M. intracellulare. Other strains of M. intracellulare, as well as M. vaccae and M. nonchromogenicum, failed to establish persistent infections in normal mice, regardless of whether they were introduced by an intravenous or subcutaneous (footpad) route. The relevance of these findings is discussed in relation to antileprosy immunity in experimental animals and in humans.

Animals

Host eicosanoid signals define a granuloma fibroblast population that coordinates mycobacterial containment.

Genetic variation at the leukotriene A4 hydrolase (LTA4H) locus is associated with tuberculosis (TB) severity and outcome. Here, we define a unique population of peripheral fibroblasts at the mycobacterial granuloma, the central immune structure in TB, whose recruitment and functions are coordinated by lta4h-dependent signals. Using single-cell profiling of zebrafish mycobacterial infections, we identify a layer of lta4h-dependent recruited fibroblasts at the granuloma's edge with mesenchymal and stem-like expression signatures, including aldh1a3 expression. Ablation of these cells compromises bacterial containment at the structure's periphery. Similarly, genetic disruption of apolipoprotein D, produced specifically in granuloma-associated fibroblasts, results in an altered eicosanoid balance, decreased inflammation, and increased dissemination of infection. In humans, this granuloma-associated fibroblast population is distinct from myofibroblasts, interacts with LTA4H-expressing macrophages, and is prominent across diverse TB granuloma types. These results link a host genetic susceptibility locus to the recruitment and function of a specialized fibroblast population that limits bacterial dissemination.

Animals

Further studies of a new pathogenic mycobacterium (M. haemophilum sp. nov.).

Mycobacterium haemophilum is an acid-fast rod-shaped organism, originally isolated from deep subcutaneous granulomata of a patient with Hodgkin's disease. Like the other two mycobacterial skin-pathogens, M. ulcerans and M. marinum, M. haemophilum has a maximum temperature for growth below 37 degrees C. Mycobacterium haemophilum is distinguished from all other species examined by its requirement of haemin for growth and its complete lack of catalase activity. Extraneous catalase cannot replace haemin as a growth factor for this organism. Mycobacterium haemophilum can also be differentiated from other species by the patterns of electrophoresis of protein extracts and by gas-liquid chromatography of saponificated and methylated lipid extracts. A monospecific-agglutinating antiserum against M. haemophilum was obtained by adsorption of an immunoserum with M. intracellulare. A number of slow-growing mycobacterial species develop on monolayers of McCoy fibroblasts, and growth on these tissue cultures can be observed much earlier than on artificial media. Mycobacterium haemophilum is characterized by exclusively intracellular development.

Aerobiosis

Atypical mycobacteria causing non-pulmonary disease in Queensland.

During the period 1971--7, the Tuberculosis Reference Laboratory in Queensland dealt with 52 isolates of atypical mycobacteria made from non-pulmonary sites under circumstances suggesting complicity in disease. Twenty-four isolates belonging to the MAIS complex were associated with lymph node infections in children. Twelve isolates belonged to the M. fortuitum-chelonei complex; most were from superficial abscesses. Five cases of M. marinum infection and 8 of M. ulcerans disease were detected.

Adolescent

Immunity to Mycobacterium leprae infections in mice stimulated by M. leprae, BCG, and graft-versus-host reactions.

Infections of mice with Mycobacterium leprae in one rear foot pad immunized them against a second infection in the other rear foot pad. Purified bacilli harvested from the first infection also produced immuniy when injection into the foot pads of previously uninfected mice. Injections of BCG afforded similar protection, but had no adjuvant effect on M. leprae. M. duvali, a cultivable mycobacterium that is reported to be more closely related antigenically to M. leprae than BCG is, provided much less protection against M. leprae challenge than BCG did. Moreover, when M. duvali was mixed with BCG, it was not any more effective than BCG alone. Graft-versus-host reactions, induced by injections of parental spleen cells into F1 hybrids, provided no protection against M. tuberculosis and M. marinum challenge. They gave moderate protection against M. leprae in one experiment but not in another with a different schedule. Allogenic spleen cells had a protective effect when injected locally into the infected foot pad. The effect produced by these injections of spleen cells was a delay in the appearance of bacterial growth; however, there was no decrease in the rate of logarithmic growth when it did appear and no reduction in the eventual plateau level.

Animals

[Research in the campaign against leprosy (author's transl)].

"Leprosy Relief through Leprosy Research" means that the results of research are made available for curing and eradicating the disease. The "Marinum Model" and the "planter test in mice" are, along with determination of serum activity in healthy test subjects, part of a complex of experiments for the assessment of the therapeutic value of an antimycobacterial substance. This replaces the "controlled studies" which, in their proper form, are scarcely possible for leprosy. With the recently developed forms of combination therapy, the duration of leprosy treatment is reduced to a few years. Because of the relationship of Mycobacterium leprae to Mycobacterium tuberculosis, certain types of combination therapy can be used in both diseases at the same time.

Animals