• Parasites, Hosts and Diseases
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• v.47 no.4
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• pp.337-344
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• 2009

In a previous study, we reported our discovery of Acanthamoeba contamination in domestic tap water; in that study, we determined that some Acanthamoeba strains harbor endosymbiotic bacteria, via our molecular characterization by mitochondrial DNA restriction fragment length polymorphism (Mt DNA RFLP). Five (29.4%) among 17 Acanthamoeba isolates contained endosymbionts in their cytoplasm, as demonstrated via orcein staining. In order to estimate their pathogenicity, we conducted a genetic characterization of the endosymbionts in Acanthamoeba isolated from domestic tap water via 16S rDNA sequencing. The endosymbionts of Acanthamoeba sp. KA/WP3 and KA/WP4 evidenced the highest level of similarity, at 97% of the recently published 16S rDNA sequence of the bacterium, Candidatus Amoebophilus asiaticus. The endosymbionts of Acanthamoeba sp. KA/WP8 and KA/WP12 shared a 97% sequence similarity with each other, and were also highly similar to Candidatus Odyssella thessalonicensis, a member of the $\alpha$-proteobacteria. The endosymbiont of Acanthamoeba sp. KA/WP9 exhibits a high degree of similarity (85-95%) with genus Methylophilus, which is not yet known to harbor any endosymbionts. This is the first report, to the best of our knowledge, to show that Methylophilus spp. can live in the cytoplasm of Acanthamoeba.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.4
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• pp.337-343
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• 2006

Trimethylamine dehydrogenase (TMADH, EC 1.5.99.7), an iron-sulfur flavoprotein that catalyzes the oxidative demethylation of trimethylamine to form dimethylamine and formaldehyde, was purified from Methylophaga sp. strain SK1. The active TMADH was purified 12.3-fold through three purification steps. The optimal pH and temperature for enzyme activity was determined to be 8.5 and $55^{\circ}C$, respectively. The $V_{max}\;and\;K_m$ values were 7.9 nmol/min/mg protein and 1.5 mM. A genomic DNA of 2,983 bp from Methylophaga sp. strain SK1 was cloned, and DNA sequencing revealed the open reading frame (ORF) of the gene coding for TMADH. The ORF contained 728 amino acids with extensive identity (82%) to that of Methylophilus methylotrophus $W_3A_1$.

• Molecules and Cells
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• v.20 no.3
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• pp.392-400
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• 2005

The genes encoding the DNA gyrase A (GyrA) and B subunits (GyrB) of Methylovorus sp. strain SS1 were cloned and sequenced. gyrA and gyrB coded for proteins of 846 and 799 amino acids with calculated molecular weights of 94,328 and 88,714, respectively, and complemented Escherichia coli gyrA and gyrB temperature sensitive (ts) mutants. To analyze the role of type II topoisomerases in the intrinsic quinolone resistance of methylotrophic bacteria, the sequences of the quinolone resistance-determining regions (QRDRs) in the A subunit of DNA gyrase and the C subunit (ParC) of topoisomerase IV (Topo IV) of Methylovorus sp. strain SS1, Methylobacterium extorquens AM1 NCIB 9133, Methylobacillus sp, strain SK1 DSM 8269, and Methylophilus methylotrophus NCIB 10515 were determined. The deduced amino acid sequences of the QRDRs of the ParCs in the four methylotrophic bacteria were identical to that of E. coli ParC. The sequences of the QRDR in GyrA were also identical to those in E. coli GyrA except for the amino acids at positions 83, 87, or 95. The $Ser^{83}$ to Thr substitution in Methylovorus sp. strain SS1, and the $Ser^{83}$ to Leu and $Asp^{87}$ to Asn substitutions in the three other methylotrophs, agreed well with the minimal inhibitory concentrations of quinolones in the four bacteria, suggesting that these residues play a role in the intrinsic susceptibility of methylotrophic bacteria to quinolones.

• Journal of Microbiology and Biotechnology
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• v.29 no.7
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• pp.1104-1116
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• 2019

In this study, we investigated the potential of using sediment bioelectrochemical systems (SBESs) for in situ treatment of the water and sediment in brackish aquaculture ponds polluted with uneaten feed. An SBES integrated into a laboratory-scale tank simulating a brackish aquaculture pond was established. This test tank and the control (not containing the SBES) were fed with shrimp feed in a scheme that mimics a situation where 50% of feed is uneaten. After the SBES was inoculated with microbial sources from actual shrimp pond sediments, electricity generation was well observed from the first experimental week, indicating successful enrichment of electrochemically active bacteria in the test tank sediment. The electricity generation became steady after 3 weeks of operation, with an average current density of $2.3mA/m^2$ anode surface and an average power density of $0.05mW/m^2$ anode surface. The SBES removed 20-30% more COD of the tank water, compared to the control. After 1 year, the SBES also reduced the amount of sediment in the tank by 40% and thus could remove approximately 40% more COD and approximately 52% more nitrogen from the sediment, compared to the control. Insignificant amounts of nitrite and nitrate were detected, suggesting complete removal of nitrogen by the system. PCR-DGGE-based analyses revealed the dominant presence of Methylophilus rhizosphaerae, Desulfatitalea tepidiphila and Thiothrix eikelboomii, which have not been found in bioelectrochemical systems before, in the bacterial community in the sediment of the SBES-containing tank. The results of this research demonstrate the potential application of SBESs in helping to reduce water pollution threats, fish and shrimp disease risks, and thus farmers' losses.