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Microbiology and Immunology 2006In order to study the diversity and community of genus Mycobacterium in polluted soils, we tried to isolate mycobacteria from 11 soil samples collected from an illegal...
In order to study the diversity and community of genus Mycobacterium in polluted soils, we tried to isolate mycobacteria from 11 soil samples collected from an illegal dumping site and 3 landfills in Japan. Using culture methods with or without Acanthamoeba culbertsoni, a total of 19 isolates of mycobacteria were obtained from 5 soil samples and 3 of them were isolated only by the co-culture method with the amoeba. Conventional biochemical tests and sequencing of the hsp65, rpoB, and 16S rRNA genes were performed for species identification of 17 of the 19 isolates. Among the 17 isolates, there was one isolate each of Mycobacterium vanbaalenii, Mycobacterium mageritense, Mycobacterium frederiksbergense, M. vanbaalenii or Mycobacterium austroafricanum, and Mycobacterium chubuense or Mycobacterium chlorophenolicum. The remaining 12 isolates could not be precisely identified at the species level. A phylogenic tree based on the hsp65 sequences indicated that 2 of the 12 isolates were novel species. In addition, 4 isolates were phylogenically close to species that degrade polycyclic aromatic hydrocarbons, which induce some cancers in humans. These results demonstrated that there were many hitherto-unreported mycobacteria in the polluted soils, and suggested that some mycobacteria might play roles in the natural attenuation and engineered bioremediation of contaminated sites with other microorganisms.
Topics: Acanthamoeba; Animals; Bacterial Proteins; Chaperonin 60; Chaperonins; DNA-Directed RNA Polymerases; Japan; Mycobacterium; Phenotype; Phylogeny; RNA, Ribosomal, 16S; Refuse Disposal; Soil Microbiology
PubMed: 16858142
DOI: 10.1111/j.1348-0421.2006.tb03821.x -
The Korean Journal of Parasitology Jun 1999To determine the pathogenicity of Acanthamoeba spp. isolated in Korea and to develop a isoenzymatic maker, the mortality rate of infected mice, in vitro cytotoxicity...
To determine the pathogenicity of Acanthamoeba spp. isolated in Korea and to develop a isoenzymatic maker, the mortality rate of infected mice, in vitro cytotoxicity against target cells and isoenzyme band patterns were observed. Five isolates of Acanthamoeba spp. (YM-2, YM-3, YM-4, YM-5, and YM-7) were used in this study as well as three reference Acanthamoeba spp. (A. culbertsoni, A. hatchetti, and A. royreba). According to the mortality rate of infected mice, Korean isolates could be categorized into three groups high virulent (YM-4), low virulent (YM-2, YM-5, YM-7) and the nonpathogenic group (YM-3). In addition, the virulence of Acanthamoeba spp. was enhanced by brain passage in mice. In the cytotoxicity assay against chinese hamster ovary cells, especially, the cytotoxicity of brain-passaged amoebae was relatively higher than the long-term cultivated ones. The zymodeme patterns of glucose-6-phosphate dehydrogenase (G6PD), malate dehydrogenase (MDH), hexokinase (HK), glutamate oxaloacetate transaminase (GOT) and malic enzyme (ME) of Acanthamoeba spp. were different among each isolate, and also between long-term cultured amoebae and brain passaged ones. In spite of the polymorphic zymodemes, a slow band of G6PD and HK, and an intermediate band of MDH were only observed in pathogenic Acanthamoeba spp., which should be used as isoenzymatic makers.
Topics: Acanthamoeba; Amebiasis; Animals; Aspartate Aminotransferases; CHO Cells; Cricetinae; Glucosephosphate Dehydrogenase; Hexokinase; Isoenzymes; Korea; Malate Dehydrogenase; Mice; Mice, Inbred ICR; Virulence
PubMed: 10388266
DOI: 10.3347/kjp.1999.37.2.85 -
Journal of Clinical Pathology Aug 1978Acanthamoeba culbertsoni was identified retrospectively in a case of amoebic meningoencephalitis, previously reported by Jager and Stamm (Lancet, 2, 1343, 1972). This is...
Acanthamoeba culbertsoni was identified retrospectively in a case of amoebic meningoencephalitis, previously reported by Jager and Stamm (Lancet, 2, 1343, 1972). This is the second report of this species causing secondary infection in man. Positive results were obtained only with anti-A. culbertsoni sera when the brain sections were stained by the indirect immunofluorescence antibody test with various antisera produced against different Acanthamoeba species. Antiserum raised against purified plasma membranes of A. culbertsoni showed once more its highly specific diagnostic value.
Topics: Adult; Antibody Specificity; Cell Membrane; Fluorescent Antibody Technique; Hartmannella; Humans; Male; Meningoencephalitis
PubMed: 357454
DOI: 10.1136/jcp.31.8.717 -
The Korean Journal of Parasitology Jun 2000In order to observe the cytotoxicity of Acanthamoeba spp., which were isolated from contact lens containers as ethiological agents for the probable amoebic keratitis in...
In order to observe the cytotoxicity of Acanthamoeba spp., which were isolated from contact lens containers as ethiological agents for the probable amoebic keratitis in Korea, the crystal violet staining method and LDH release assay were carried out. In the crystal violet staining method, among eight contact lens container isolates, isolate 3 (Acanthamoeba KA/LS5) showed 83.6% and 81.8% of cytotoxicity, and isolate 7 (Acanthamoeba KA/LS37) showed 28.2% and 25.1% of cytotoxicity, in 1 mg/ml and 0.5 mg/ml lysate treatments, respectively. Acanthamoeba culbertsoni and A. healyi showed 84.0% and 82.8% of cytotoxicity. Similar results were observed in A. castellanii and A. hatchetti which showed 83.6% and 75.5% of cytotoxicity. Acanthamoeba royreba and A. polyphaga showed 9.0% and 1.7% of cytotoxicity. In the LDH release assay, isolate 3 (20.4%) showed higher cytotoxicity than other isolates in 1 mg/ml lysate treatment. The results provide that at least isolate 3 has the cytotoxic effect against CHO cells and seems to be the pathogenic strain.
Topics: Acanthamoeba; Animals; CHO Cells; Cell Survival; Contact Lenses; Cricetinae; Equipment Contamination; Gentian Violet; Humans; Korea; L-Lactate Dehydrogenase; Staining and Labeling
PubMed: 10905072
DOI: 10.3347/kjp.2000.38.2.99 -
Journal of Clinical Microbiology Apr 1985A case of amoebic meningitis, presumably primary, was encountered in the Christian Medical College Hospital, Vellore, South India, in November 1983. The patient, a...
A case of amoebic meningitis, presumably primary, was encountered in the Christian Medical College Hospital, Vellore, South India, in November 1983. The patient, a 40-year-old man, had cerebrospinal fluid rhinorrhea before the meningitis developed. Acanthamoeba culbertsoni was repeatedly demonstrated in and cultured from the cerebrospinal fluid. The patient responded dramatically to a combination therapy of penicillin and chloramphenicol.
Topics: Adult; Amebiasis; Animals; Follow-Up Studies; Hartmannella; Humans; Male; Meningitis; Mice
PubMed: 3988911
DOI: 10.1128/jcm.21.4.666-667.1985 -
Clinical and Diagnostic Laboratory... Sep 1994Prominent antigens of pathogenic and nonpathogenic free-living amoebae were identified by using polyclonal rabbit immune sera in immunoblot assays. The intent was to...
Prominent antigens of pathogenic and nonpathogenic free-living amoebae were identified by using polyclonal rabbit immune sera in immunoblot assays. The intent was to determine if prominent epitopes identified with rabbit immune sera could also be recognized by human sera. With rabbit sera, the development of immunoreactive bands was restricted to molecular masses of greater than 18.5 kDa for Naegleria, Hartmannella, and Vahlkampfia antigens. Two or more broad bands of less than 18.5 kDa were prominent features in three different Acanthamoeba species. Few cross-reactive antibodies could be detected between representative species of the three different subgroups of Acanthamoeba. Naegleria antigen was likewise serologically distinct, as were Hartmannella and Vahlkampfia antigens. The relative lack of cross-reacting antibodies between the pathogenic amoebae suggested that i would be desirable to use a panel of amoebic antigens to represent the range of serologically distinct antigens for assessing reactive antibodies in human sera. In pooled human sera (10 serum specimens per pool), the appearance of minimally reactive bands ranging from 32.5 to 106 kDa was a common feature of all six antigens. A prominent band of less than 18.5 kDa was identified in the Acanthamoeba culbertsoni antigen lane in 2 of the 10 human serum specimen pools. When sera from each of the two groups were tested individually by immunoblotting, the reaction with A. culbertsoni antigen could be associated with one individual. By using a panel of amoebic antigens, this method could prove useful in recognizing undiagnosed amoebic infections by revealing specific reactive antibodies.
Topics: Acanthamoeba; Adult; Animals; Antibody Specificity; Antigens, Protozoan; Blotting, Western; Cross Reactions; Electrophoresis, Polyacrylamide Gel; Epitopes; Fluorescent Antibody Technique; Hartmannella; Humans; Infant; Naegleria fowleri; Rabbits
PubMed: 8556491
DOI: 10.1128/cdli.1.5.493-499.1994 -
The Korean Journal of Parasitology Dec 1995Genetic status of Acanthamoeba spp. were tested on the basis of random amplified polymorphic DNA (RAPD) marker analysis. Four previously established Acanthamoeba...
Genetic status of Acanthamoeba spp. were tested on the basis of random amplified polymorphic DNA (RAPD) marker analysis. Four previously established Acanthamoeba species, 4 Korean isolates of Acanthamoeba sp., and one American isolate of Acanthamoeba sp. were analyzed by RAPD-PCR using an arbitrary decamer primers. Amplification products were fractionated by agarose gel electrophoresis and stained by ethidium bromide. Eighteen primers produced DNA amplification profiles revealing clear differences among 4 species. Nine of them also produced DNA amplification profiles which included some isolate-specific amplification products. On the basis of amplified fragments by 18 primers, the pairwise similarity indices between A. culbertsoni and other species (i.e., A. hatchetti, A. triangularis, A. polyphaga) were 0.300, 0.308, and 0.313, respectively. Similarity index between A. hatchetti and A. triangularis was 0.833. The mean similarity index among the 3 Korean isolates (YM-2, -3, -4) was 0.959 and 0.832 among them and 2 other species (A. hatchetti and A. triangularis). The mean similarity index among YM-5 and other Korean isolates (YM-2, -3, -4) was 0.237. However, the similarity index between YM-5 and A. culbertsoni was 0.857, which suggests that YM-5 is genetically more similar to A. culbertsoni than other Korean isolates. Phenogram reconstructed by UPGMA method revealed that there are two groups: one group consists of A. hatchetti, A. triangularis, and 3 Korean isolates (YM-2, -3, -4), and the other group consists of A. culbertsoni, A. polyphaga, HOV, and YM-5.
Topics: Acanthamoeba; Animals; Korea; Polymerase Chain Reaction; Random Amplified Polymorphic DNA Technique
PubMed: 8591012
DOI: 10.3347/kjp.1995.33.4.341 -
The Korean Journal of Parasitology Jun 1993Acanthamoeba culbertsoni is a pathogenic free-living amoeba causing primary amoebic meningoencephalitis (PAME) in human and mouse. Several reports on the immune...
Acanthamoeba culbertsoni is a pathogenic free-living amoeba causing primary amoebic meningoencephalitis (PAME) in human and mouse. Several reports on the immune responses in mice with this amoebic infection have been published, but the effects of transferred passive immunity on the active immunization in offspring mice have not been demonstrated. This experiment was done to observe the effect of active immunization with Acanthamoeba culbertsoni in mice born to immune mothers. Acanthamoeba culbertsoni was cultured in the CGV medium axenically. Female BALB/c mice weighing about 20g were immunized through the intraperitoneal injection of Acanthamoeba culbertsoni trophozoites 1 x 10(6) each three times at the interval of one week. Offspring mice were immunized two times. The mice were inoculated intranasally with 1 x 10(4) trophozoites under secobarbital anesthesia. There was a statistical difference in mortality between the transferred immunity group and the active immunization group. Statistical differences were not demonstrated in antibody titer between both groups. But L3T4+ T cell/Ly2+ T cell ratio was increased in the transferred immunity group more than active immunization group of the offspring mice at the age of 5 weeks. There was no differences statistically in mortality between both groups. It was recognized that active immunization in offspring mice born to immune mother could modulate the immune status according to the time of immunization.
Topics: Acanthamoeba; Amebiasis; Animals; Antibodies, Protozoan; Female; Immunity, Maternally-Acquired; Immunization Schedule; Mice; Mice, Inbred BALB C; Vaccination
PubMed: 8343458
DOI: 10.3347/kjp.1993.31.2.157 -
Infection and Immunity Jun 2004Legionella dumoffii is one of the common causes of Legionnaires' disease and is capable of replicating in macrophages. To understand the mechanism of survival within...
Legionella dumoffii is one of the common causes of Legionnaires' disease and is capable of replicating in macrophages. To understand the mechanism of survival within macrophages, transposon mutagenesis was employed to isolate the genes necessary for intracellular growth. We identified four defective mutants after screening 790 transposon insertion mutants. Two transposon insertions were in genes homologous to icmB or dotC, within dot/icm loci, required for intracellular multiplication of L. pneumophila. The third was in a gene whose product is homologous to the 17-kDa antigen forming part of the VirB/VirD4 type IV secretion system of Bartonella henselae. The fourth was in the djlA (for "dnaj-like A") gene. DjlA is a member of the DnaJ/Hsp40 family. Transcomplementation of the djlA mutant restored the parental phenotype in J774 macrophages, A549 human alveolar epithelial cells, and the amoeba Acanthamoeba culbertsoni. Using confocal laser-scanning microscopy and transmission electron microscopy, we revealed that in contrast to the wild-type strain, L. dumoffii djlA mutant-containing phagosomes were unable to inhibit phagosome-lysosome fusion. Transmission electron microscopy also showed that in contrast to the virulent parental strain, the djlA mutant was not able to recruit host cell rough endoplasmic reticulum. Furthermore, the stationary-phase L. dumoffii djlA mutants were more susceptible to H2O2, high osmolarity, high temperature, and low pH than was their parental strain. These results indicate that DjlA is required for intracellular growth and organelle trafficking, as well as bacterial resistance to environmental stress. This is the first report demonstrating that a single DjlA-deficient mutant exhibits a distinct phenotype.
Topics: Acanthamoeba; Amino Acid Sequence; Animals; Bacterial Proteins; Cell Line; DNA Transposable Elements; Epithelial Cells; HSP40 Heat-Shock Proteins; Heat-Shock Proteins; Heat-Shock Response; Humans; Legionella; Lysosomes; Macrophages; Mice; Microscopy, Confocal; Microscopy, Electron; Mutagenesis, Insertional; Phagosomes
PubMed: 15155669
DOI: 10.1128/IAI.72.6.3592-3603.2004 -
The Korean Journal of Parasitology Sep 1993The purpose of this observation was to investigate the natural killer cell activities in mice infected with pathogenic free-living amoeba, Naegleria fowleri and...
The purpose of this observation was to investigate the natural killer cell activities in mice infected with pathogenic free-living amoeba, Naegleria fowleri and Acanthamoeba culbertsoni according to the infection doses, and infected with non-pathogenic free-living amoeba, Naegleria gruberi. The natural killer cell activity was examined by means of target binding capacity, active NK cell and maximum recycling capacity of the mice after inoculating free-living amoebae with low and high doses. The mice infected with 1,103, 1,105 A. culbertsoni trophozoites showed mortality rates of 6.9% and 65.5%, respectively. The mice infected with 1,104, 1,105 N. fowleri trophozoites showed mortality rates of 5.9% and 72.2%, respectively. The NK cell activities in all experimental groups increased significantly on day 1 after infection as compared with control group, and then remarkably declined thereafter, there was no difference of the cytotoxic activity of the NK cells in mice among inoculation doses of pathogenic free-living amoebae. The target binding capacities of NK cells and percentages of activated NK cells in mice infected with pathogenic free-living amoebae were significantly increased a day after infection, as compared with control group. There was no difference of the maximal recycling capacities of NK cells in all experimental groups as compared with control group. There was significant difference in the cytotoxic activity and single cell cytotoxicity of NK cells between the experimental groups infected with pathogenic free-living amoebae and that infected with non-pathogenic free-living amoebae.
Topics: Acanthamoeba; Amebiasis; Animals; Cytotoxicity, Immunologic; Killer Cells, Natural; Mice; Mice, Inbred C3H; Naegleria fowleri
PubMed: 8241083
DOI: 10.3347/kjp.1993.31.3.239