Tsukamurella Inchonensis Research Articles

<i>Tsukamurella Inchonensis</i> Isolated from Catheter of an Ecuadorian Patient with Hemodialysis

<i>Tsukamurella Inchonensis</i> Isolated from Catheter of an Ecuadorian Patient with Hemodialysis

Mucosal Immunoregulatory Properties of Tsukamurella inchonensis to Reverse Experimental Food Allergy.

The intestinal mucosa is lined by epithelial cells, which are key cells to sustain gut homeostasis. Food allergy is an immune-mediated adverse reaction to food, likely due to defective regulatory circuits. Tsukamurella inchonensis is a non-pathogenic bacterium with immunomodulatory properties. We hypothesize that the anti-inflammatory effect of dead T. inchonensis on activated epithelial cells modulates milk allergy through the restoration of tolerance in a mouse model. Epithelial cells (Caco-2 and enterocytes from mouse gut) and macrophages were stimulated with T. inchonensis and induction of luciferase under the NF-κB promoter, ROS and cytokines production were studied. Balb/c mice were mucosally sensitized with cow´s milk proteins plus cholera toxin and orally challenged with the allergen to evidence hypersensitivity symptoms. After that, mice were orally administered with heat-killed T. inchonensis as treatment and then challenged with the allergen. The therapeutic efficacy was in vivo (clinical score and cutaneous test) and in vitro (serum specific antibodies and cytokines-ELISA, and cell analysis-flow cytometry) evaluated. Heat-killed T. inchonensis modulated the induction of pro-inflammatory chemokines, with an increase in anti-inflammatory cytokines by intestinal epithelial cells and by macrophages with decreased OX40L expression. In vivo, oral administration of T. inchonensis increased the frequency of lamina propria CD4+CD25+FoxP3+ T cells, and clinical signs were lower in T. inchonensis-treated mice compared with milk-sensitized animals. In vivo depletion of Tregs (anti-CD25) abrogated T. inchonensis immunomodulation. In conclusion, these bacteria suppressed the intestinal inflammatory immune response to reverse food allergy.

The effects of heat-killed Tsukamurella inchonensis on intestinal morphology and humoral immune responses of broiler chickens.

Background and Objectives:Tsukamurella species are Gram-positive rods that exist in a broad range of environments. In this study, the efficacy of heat-killed Tsukamurella inchonensis on growth performance, intestinal morphology, and humoral immune responses of broiler chicken was evaluated.Materials and Methods:Ross broiler chicks in the cage were randomly allocated to five groups. Trail diets were prepared by adding 106 cells per bird of heat-killed T. inchonensis into the basal trading diet for group 1 continuously dosed for 24 h from day 1 to day 13, and for group 2, 24 h on days 1 to 5; 8; 9, 12 and 13. Group 3 was received 106 bacteria as a subcutaneous injection on days 1, 6, and 12. Groups 4 and 5 were not received T. inchonensis during the experiment period.Results:Feed intake (FI) and feed conversion ratio (FCR) were not altered by different delivery methods of T. inchonensis supplementation. The pulsed dosed in feed tended to provide higher body weight gain (BWG) than the negative control groups. T. inchonensis treatments, never less of the ways of delivery, boosted (P<0.05) the antibody titers to Newcastle disease virus (NDV), and avian influenza (AI) (H9N2) virus, especially when broiler chickens treated with pulse dosed in the feed. The most significant intestinal development (p<0.05) was observed between groups 1 and 2. There were no significant differences in the thymus, liver, and bursa of Fabricius relative weight. Still, there were significant increases in the relative weight of spleen on day 14 in vaccinated chickens treated with T. inchonensis pulse dosed.Conclusion:It seems that the supplementation of T. inchonensis in the broiler diet can improve intestinal morphology and humoral immune response, which was represented by increased antibody response to NDV, and AI vaccines significantly, but it cannot affect FI and FCR.

The Beneficial Effects of Actinomycetales Immune Modulators in the Pancreas of Diabetic Rats.

Purpose: Type 1 diabetes mellitus (T1DM) has dramatically increased in recent years, especially in young people, and limits the life quality of the patients involved. Thus, many researchers are performing extensive studies to find alternative treatments for DM.Methods: Here, we evaluated the improvement effects of the heat-killed Actinomycetales species, including Gordonia bronchialis, and Tsukamurella inchonensis in streptozotocin (STZ)- diabetic rats by biochemical, immunological, and histopathological examinations.Results: The present findings exhibited a dramatic and progressive alteration in the serum levels of interleukin-6 (IL-6), IL-10 and tumor necrosis factor-α (TNF-α) in the diabetic group, which were related to the blood glucose and insulin levels, oxidative stress defense (evaluated by TAC and MDA activities), and the pancreas biochemical indicators (such as amylase and lipase). More importantly, the present results were consistent with the histopathological findings, which included cellular degeneration, vascular congestion, hemorrhage, focal necrosis associated with mononuclear cell infiltration. Interestingly, all of the diabetic changes in the blood serum and tissues improved remarkably in the treated groups by Actinomycetales species.Conclusion: Surprisingly, most of the current diabetic complications effectively attenuated after oral administration of both Actinomycetales species, particularly with a high dose of T. inchonensis. Thus, it is concluded that the heat-killed Actinomycetales species can prevent and improve the progression of T1DM and its various complications profoundly.

Identification of Petroleum Degrading Bacteria and Construction of Petroleum Degrading Agent

The aim of this study is to construct petroleum degrading agent (PDA) which can effectively degrade oil. By enrichment, domestication, and separation of culture from the soil sample of Qianjiang Guanghua Oilfield, the pure culture of three petroleum degrading bacteria G-40, G-53, and G-94 was identified from the medium supplemented with oil, which served as the sole source of carbon. The species of G-40 and G-53 were preliminarily identified and classified by morphological observation, physiological and biochemical determination, and sequence analyses of 16S rDNA. The species of G-94 was preliminarily identified and classified by morphological observation, physiological and biochemical determination, and sequence analyses of ITS rDNA. The optimal inoculation proportion of these three bacteria strains and bran proportion in composition of PDA were determined through orthogonal test. G-40, G-53, and G-94 were isolated and identified as Brevibacillus laterosporus, Tsukamurella inchonensis, and Candida tropicalis, respectively. To construct petroleum degrading agent, the optimum inoculation proportion of the three bacteria strains was A1B3C3 (G-40:G-53:G-94 = 1:4:4); and the optimum proportion of bran was D1E1F2 (soybean meal:corn flour:bran = 1:1:2). The oil removal rate of the constructed petroleum degrading agent reached to 42.32% on day 10 under the optimal proportion of bacteria inoculation and bran composition. Petroleum degrading bacteria can effectively degrade petroleum for its own growth. This study identified three petroleum degrading bacteria strains and proposed a petroleum degrading agent by studying the optimal inoculation proportion of the three bacterial strains and the accompanying bran. Our research could provide potential microbial resources for bioremediation of petroleum-contaminated soil.

Heat-killed Tsukamurella inchonensis reduces lipopolysaccharide-induced inflammatory responses in activated murine peritoneal macrophages

Tsukamurella inchonensis (T. inchonensis) is an aerobic species of Actinomycetales which has immunomodulatory activities when used as a suspension of killed bacilli. Here, the effects of T. inchonensis on lipopolysaccharide-induced inflammatory responses in mouse peritoneal macrophages have been examined. Peritoneal macrophages were harvested by lavaging with ice cold phosphate-buffered saline. Macrophages acquired from mice treated with different doses of T. inchonensis for seven days were cultured with 20 U/ml interferon-γ and 10 µg/ml lipopolysaccharide for in vivo assays. Nitrite levels were measured by using the diazotization method based on the Griess reaction, an indirect technique to determine nitric oxide (NO) production. T. inchonensis inhibited lipopolysaccharide-stimulated NO production in mouse peritoneal macrophages from mice previously exposed to concentrations of 108 and 5 × 10&lt;sup&gt;7&lt;/sup&gt; CFU per flask. Also, T. inchonensis decreased lipopolysaccharide-induced production of pro-inflammatory cytokines, including interleukin-6 and tumor necrosis factor-α. Thus, it can be concluded that T. inchonensis is a powerful inhibitor of lipopolysaccharide-induced NO production in activated murine macrophages, and T. inchonensis may be useful as a novel agent for chemoprevention in inflammatory diseases.

Korean indigenous bacterial species with valid names belonging to the phylum Actinobacteria.

To understand the isolation and classification state of actinobacterial species with valid names for Korean indigenous isolates, isolation source, regional origin, and taxonomic affiliation of the isolates were studied. At the time of this writing, the phylum Actinobacteria consisted of only one class, Actinobacteria, including five subclasses, 10 orders, 56 families, and 330 genera. Moreover, new taxa of this phylum continue to be discovered. Korean actinobacterial species with a valid name has been reported from 1995 as Tsukamurella inchonensis isolated from a clinical specimen. In 1997, Streptomyces seoulensis was validated with the isolate from the natural Korean environment. Until Feb. 2016, 256 actinobacterial species with valid names originated from Korean territory were listed on LPSN. The species were affiliated with three subclasses (Acidimicrobidae, Actinobacteridae, and Rubrobacteridae), four orders (Acidimicrobiales, Actinomycetales, Bifidobacteriales, and Solirubrobacterales), 12 suborders, 36 families, and 93 genera. Most of the species belonged to the subclass Actinobacteridae, and almost of the members of this subclass were affiliated with the order Actinomycetales. A number of novel isolates belonged to the families Nocardioidaceae, Microbacteriaceae, Intrasporangiaceae, and Streptomycetaceae as well as the genera Nocardioides, Streptomyces, and Microbacterium. Twenty-six novel genera and one novel family, Motilibacteraceae, were created first with Korean indigenous isolates. Most of the Korean indigenous actionobacterial species were isolated from natural environments such as soil, seawater, tidal flat sediment, and fresh-water. A considerable number of species were isolated from artificial resources such as fermented foods, wastewater, compost, biofilm, and water-cooling systems or clinical specimens. Korean indigenous actinobacterial species were isolated from whole territory of Korea, and especially a large number of species were from Jeju, Gyeonggi, Jeonnam, Daejeon, and Chungnam. A large number of novel actinobacterial species continue to be discovered since the Korean government is encouraging the search for new bacterial species and researchers are endeavoring to find out novel strains from extreme or untapped environments.

Bacteria Associated with Larvae and Adults of the Asian Longhorned Beetle (Coleoptera: Cerambycidae)

Bacteria representing several genera were isolated from integument and alimentary tracts of live Asian longhorned beetle, Anaplophora glabripennis (Motschulsky), larvae and adults. Insects examined were from infested tree branches collected from sites in New York and Illinois. Staphylococcus sciuri (Kloos) was the most common isolate associated with adults, from 13 of 19 examined, whereas members of the Enterobacteriaceae dominated the isolations from larvae. Leclercia adecarboxylata (Leclerc), a putative pathogen of Colorado potato beetle, Leptinotarsa decemlineata (Say), was found in 12 of 37 larvae examined. Several opportunistic human pathogens, including S. xylosus (Schleifer and Kloos), S. intermedius (Hajek), S. hominis (Kloos and Schleifer), Pantoea agglomerans (Ewing and Fife), Serratia proteamaculans (Paine and Stansfield) and Klebsiella oxytoca (Flugge) also were isolated from both larvae and adults. One isolate, found in 1 adult and several larvae, was identified as Tsukamurella inchonensis (Yassin) also an opportunistic human pathogen and possibly of Korean origin.. We have no evidence that any of the microorganisms isolated are pathogenic for the Asian longhorned beetle.

Successful Immunotherapy of Canine Flea Allergy with Injected Actinomycetales Preparations

Can heat-killed, borate-buffered suspensions of Gordonia bronchialis, Rhodococcus coprophilus or Tsukamurella inchonensis be used to treat canine flea allergy? Organisms cultured on Sauton's medium into stationary phase were autoclaved in borate-buffered saline and stored at 10 mg wet weight/ml. Intradermal injections of 0.1 ml containing 1 mg of bacilli were administered on the first and 20th days of the study. G. bronchialis and R. coprophilus were most effective in a pilot study of a small number of dogs with flea allergy. A larger number of affected dogs were then randomized to receive placebo or either of the two selected reagents. The extent and severity of allergic signs and symptoms were scored and blood samples were collected just before the first injection and 28 days after the second. Both selected reagents reduced the extent and severity of lesions (p < 0.001) and reduced scratching. Eosinophil numbers were reduced (p < 0.0001) between the first and second assessment. Injections of G. bronchialis or R. coprophilus effectively reduce the signs and symptoms of flea allergy in dogs.

Immunotherapy for the Prevention of Myointimal Hyperplasia After Experimental Balloon Injury of the Rat Carotid Artery

We assessed the effect of novel immunotherapeutic heat-killed bacterial (Actinomycetales) preparations on the development of myointimal hyperplasia (MIH) in a rat carotid balloon trauma model and the effect on the immune response by measuring the expression of interferon gamma (IFN-gamma; (Th1) and interleukin 4 (IL-4; Th2). There was a significant reduction (P < .001) in intima/media ratios (mean +/- SEM) in the rats treated by immunomodulation (0.52 +/- 0.03 Gordonia bronchialis, 0.60 +/- 0.03 Rhodococcus coprophilus, 0.43 +/- 0.03 Tsukamurella inchonensis, 0.37 +/- 0.03 Mycobacterium vaccae), in comparison with untreated controls (0.91 +/- 0.05). Postballoon trauma G bronchialis increased messenger RNA (mRNA) IFN-gamma (P < .02) and reduced mRNA IL-4 (P < .05). R coprophilus, T inchonensis, and M vaccae significantly increased production of mRNA IFN-gamma (P < .001). R coprophilus and M vaccae also decreased production of mRNA IL-4 (P < .05, P < .01). Treatment with heat-killed Actinomycetales inhibits MIH through a combination of enhanced Th1 and attenuated Th2 response. Immunomodulation may provide a novel therapeutic option to prevent restenosis.

Tsukamurella tyrosinosolvens sp. nov.

Chemotaxonomic and 16S ribosomal DNA sequence analyses of four bacterial isolates from blood cultures from patients with cardiac pacemaker implants and sputa of patients with chronic lung infections clearly demonstrated that these bacteria belong to the genus Tsukamurella. DNA-DNA hybridization data, as well as the physiological characteristics of the isolates, indicate that they are closely related and belong to a single species that differs from previously described members of the genus Tsukamurella. The name Tsukamurella tyrosinosolvens sp. nov. is proposed for these isolates, and the new species is represented by strain IMMIB D-1397T (= DSM 44234T). Strain IMMIB D-1397T exhibits 53.4, 53.5, and 54.7% DNA-DNA relatedness to Tsukamurella paurometabola DSM 20162T, Tsukamurella inchonensis DSM 44067T, and Tsukamurella pulmonis DSM 44142T, respectively.

Tsukamurella pulmonis sp. nov.

Chemotaxonomic and 16S ribosomal DNA sequence analyses of an isolate from the sputum of a patient with a mycobacterial lung infection clearly delineated a new species of the genus Tsukamurella. This new species can be defined on the basis of genotypic and phenotypic data. The name Tsukamurella pulmonis sp. nov. is proposed for this organism; the type strain is IMMIB D-1321T (= DSM 44142T). This isolate shows 44.2 and 36.2% DNA relatedness to Tsukamurella paurometabola DSM 20162T (T = type strain) and Tsukamurella inchonensis DSM 44067T, respectively.

Tsukamurella inchonensis sp. nov.

Chemotaxonomic and genomic 16S ribosomal DNA sequence analyses of two isolates obtained from two different clinical materials clearly delineated a new species of the genus Tsukamurella. This new species can be identified by its 16S ribosomal DNA similarity values, as well as its physiological characteristics. The name Tsukamurella inchonensis sp. nov. is proposed for these isolates, which are represented by strain IMMIB D-771T (= DSM 44067T) (T = type strain). This strain exhibits only 45% DNA relatedness to Tsukamurella paurometabola.