Evaluation of koji prepared with various molds for mirin-making

Koji is one of the raw materials used for mirin-making, and it is traditionally prepared with Aspergillus oryzae. To improve productivity and to be available for a variety of mirin, various koji were prepared with A. niger, A. awamori, A. usamii mut. shiro-usamii, and Rhizopus oligosporus and examined for possible to use in mirin-making. The degrees of digestion of the starch and protein in various koji were at its highest with either koji prepared with A. oryzae at pH 7.0 or with A. usamii mut. shiro-usamii at pH 3.0. The degree of digestion of the protein in koji made with A. usamii mut. shiro-usamii was decreased in koji digestion at higher temperature (at over 50°C) compared to that in koji made with A. oryzae. We examined the ratio of koji to steamed glutinous rice to identify the optimum value for mirin production. The optimum ratios for digestion of starch in the mash with koji of A. oryzae and A. usamii mut. shiro-usamii were 0.10 and 0.15, respectively. These values were in agreement with the 0.10–0.25 established by experience. Therefore, of the five strains tested, A. usamii mut. shiro-usamii, among the tested molds was the most suitable strain for preparing koji in mirin-making without decreasing productivity, while lending new qualities to the mirin. The koji prepared with A. oryzae had a good smell but the koji prepared with A. usamii mut. shiro-usamii had a little mold smell. Then if this little mold smell could be improved, this koji can be used in mirin-making. The koji prepared with A. niger contained the most citric acid of all strains but the digestivities of starch and protein in the koji was less than those of koji prepared with A. oryzae and A. usamii mut. shiro-usamii. The digestivities of starch and protein in koji prepared with A. awamori were the lowest values. In 35% alcohol, its productivities in mirin-making seemed to be low. Because of the highest content of lactic acid, the koji prepared with R. oligosporus seemed to be suitable for mirin-making. Then if its productivity could be improved, this koji can be used in mirin-making.     

Some heterostrophic gastropods from Triassic St. Cassian Formation with a discussion on the classification of the Allogastropoda

Fifteen species of Heterostropha are described, 12 of them for the first time. All are newly interpreted with regard to their taxonomic relation to fossil and living gastropods. The Streptacidoidea with long Paleozoic history are represented in the Late Triassic St. Cassian Formation by several genera that can be differentiated into four families. The Ebalidae are represented byEbala, with smooth protoconch, Cassianebalidae byCassianebala andLoxebala with axially ornamented protoconch. The Donaldinidae of St. Cassian are represented by one species ofDonaldina and two ofNeodonaldina that stand in the continuation of Paleozoic species ofDonaldina. Architectonicoidea with shells coiled in a plane and Valvatoidea appear in the St. Cassian fauna without known Paleozoic relation. In the former superfamily the Architectonicidae can be recognized in the genusRinaldoconchus with two species. Cassianaxidae withCassianaxis, Amphitomariidae withAmphitomaria, Stuoraxidae withStuoraxis andAmpezzogyra have a sinistral protoconch and planispirally coiled dextral teleconchs. They all resemble different modern species that have similarly small shells. Modern Hyalogyrinidae have withAlexogyra a new representative from the Triassic. The Valvatoidea are represented with the generaCarboninia andBandellina of the Cornirostridae in the Triassic representatives. The relation of described species in the system of the Heterostropha is discussed.     

Intertidal zonation of prosobranch gastropods: Analysis of densities of four co-existing species

The densities of populations of Nerita atramentosa Reeve, Austrocochlea constricta Lamarck, Bembiciuin nanum (Lamarck), and Cellana tramoserica (Sowerby) were recorded in 0.25 m² quadrats on landward and seaward halves of a sheltered and a moderately exposed rock platform from June 1972 to June 1973. The data have been subjected to variance and multiple regression analysis. Differences between densities of each species on the two shores and in the two areas of each shore are discussed with respect to the regressions on littoral height. Densities of Nerita on the sheltered shore, Austrocochlea and Bembicium on both shores, and Cellana, on the exposed shore, increased with height. The density of Cellana on the sheltered shore decreased as height increased. The density differences in the landward and seaward areas on the two shores were attributable to the effect of height for Nerita and Austrocochlea. Height was the major variable affecting the density of these two species on the sheltered shore. Bembicium and Cellana densities on both shores, and Austrocochlea density on the exposed shore, are not primarily determined by height.On the sheltered shore, Nerita and Cellana increased in density with increasing cover of water in rock pools. On the exposed shore, Bembicium and Cellana became sparser with increasing water cover. Bembicium on both shores increased in density with increasing cover of the alga Peyssonelia, as did Cellana on the exposed shore. Cellana and Austrocochlea tended to increase in density with Nerita on the sheltered shore. Cellana tended to be sparse where Bembicium was dense on the exposed shore. The biological effects of substrata and other species on the densities of each species remain unexplained.Significant time effects were found for variations in the densities of Nerita and Cellana on the sheltered shore and of Austrocochlea and Cellana on the exposed shore. The seasonal change in density of Nerita was parabolic with a minimum in summer, and is consistent with the known period of recruitment of this species. The lack of significant time effects on density of Austrocochlea on the sheltered shore is consistent with the known continuous recruitment of this species. The declining density of Austrocochlea on the moderately exposed shore was found, by size-frequency analysis, to be due to reduced recruitment compared with surrounding shores. The density of Cellana on both shores showed seasonal changes, with a maximum in summer, which is consistent with the known period of recruitment in Cellana. The lack of seasonal changes in density of Bembicium is briefly discussed.This type of multifactor analysis identifies biological interactions affecting variation in density of these species. It also corroborates the results of other ecological investigations and determines the relative importance of a variety of physical and biological factors in the distribution of density of intertidal gastropods.     

Use of koji prepared with a high citric acid producing mutant of Aspergillus usamii as a raw material for Saké brewing

There is a possibility of developing a new kind of saké in which the refreshing sour taste of citric acid is introduced. In this study, we bred a new mutant of Aspergillus usamii mut. shiro-usamii that produced much citric acid. The koji prepared with the mutant contained about 20 mg of citric acid per gram of dry koji, twice that of the koji of the parental strain. The activities of a-amylase, glucoamylase, and acidic protease in the koji prepared with the mutant were 82%, 94%, and 95%, respectively, those of the parental strain. Using this koji with the mutant, saké was produced. The levels of citric acid and isoamyl acetate were 5.1 and 1.4 times, respectively, those of saké prepared with koji of A. oryzae. Sensory tests indicated that saké made with koji with the mutant was refreshingly sour, with a good aroma.     

Reaction of cytochrome c in the electron-transport chain of Paracoccus denitrificans

The reaction of the cytochrome c oxidase (ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1) of Paracoccus denitrificans cytoplasmic membranes with the endogenous cytochrome c of the membranes was studied, as well as its interaction with added exogenous cytochrome c from P. denitrificans or bovine heart. The polarographic method was employed, using N,N,N′,N′-tetramethyl-p-phenylenediamine plus ascorbate to reduce the cytochrome c. We found that overall electron transport can proceed maximally while the cytochrome c remains membrane bound; NADH or succinoxidase activities were not inhibited by the addition of substances which bind the P. denitrificans cytochrome c strongly. In contrast to our observations with the spectrophotometric method (Smith, L., Davies, H.C. and Nava, M.E. (1976) Biochemistry 15, 5827–5831), in the polarographic assays the membrane-bound oxidase reacts with about equal rapidity with exogenous bovine and P. denitrificans cytochromes c. The reaction of the oxidase with the endogenous cytochrome c proceeds at high rates and preferentially to that with exogenous cytochrome c; the reaction with the latter, but not the former is inhibited by positively charged poly(l-lysine). The cytochrome c and the oxidase appear to be very closely associated on the membrane.     

Cryptosporidium parvum: the course of Cryptosporidium parvum infection in C57BL/6 mice co-infected with the nematode Heligmosomoides bakeri

The effects of Heligmosomoides bakeri infection on the course of a concurrent Cryptosporidium parvum infection were studied in C57BL/6 mice. Mice were initially infected with 80 L₃ of H. bakeri and then challenged with 10⁴ oocysts of C. parvum, administered during the patent period of the nematode infection (28 day post H. bakeri infection). The number of C. parvum oocysts excreted in the feces and the number of adult H. bakeri in the small intestine were monitored during the experiment. Concurrent H. bakeri infection resulted in a prolonged course of infection with C. parvum. The intensities of both parasite infections were higher in co-infections. We also investigated the cellular immune response at 14 and 42 days post infection C. parvum. During infection with C. parvum there was an increase in production of IFN-γ and IL-12 but co-infection with H. bakeri inhibited IFN-γ secretion. The present study is the first to demonstrate that infection with H. bakeri markedly exacerbates the intensity of a concurrent C. parvum infection in laboratory mice and also affects immune effectors mechanisms in co-infection with H. bakeri.     

Two aberrant mercury resistance transposons in the Pseudomonas stutzeri plasmid pPB

The two mer operons of the Pseudomonas stutzeri OX plasmid pPB and their flanking regions have been sequenced and found to be part of two aberrant transposons. The narrow spectrum mer operon is almost identical to that of Tn501, but is associated with the remnants of Tn5053 tni genes rather than the Tn501 transposition module. The broad spectrum mer operon shows an overall homology with that of Tn5053, but differs from it in the presence of a merB gene, absent in Tn5053, and a merC gene instead of a merF. The pPB broad spectrum mer operon is associated with an incomplete Tn5053-like transposition module and with the Tn501 tnp genes, which are proximal, respectively, to the end and to the beginning of the mer operon. A hypothesis about pPB evolution is presented.     

The male antigen. II. Regulation of the primary and secondary responses to H-Y by H-2 associated genes

The primary response of females of ten inbred mouse strains to the male antigen (H-Y) was investigated by transfer of peritoneal exudate cells (PEC). Three distinct classes of reactivity were seen. Early primary response to H-Y was associated with H-2 haplotypes b and s; intermediate response with H-2 haplotypes, k, d, i, and h; and late or absent response with H-2 haplotypes a and f. The failure of A.CA (H-2^f) females to mount a detectable primary response against syngeneic male cells was not due to the lack of the antigen from A.CA male cells. The ability of (A.CA × B10)F₁ hybrid females to respond to the male antigen demonstrated the dominant nature of the B10 (H-2^b) response and excluded the possibility that A.CA females possess a dominant self-antigen cross-reactive with H-Y. The secondary response of eight inbred strains was investigated; at least three distinct levels of reactivity were apparent. The speed of the secondary response was associated with the various H-2 haplotypes in the same way as the primary response. The implications of differential strain reactivity, background effect, and the association of Ir-1 with response to H-Y are discussed.     

In vitro Effects of Prebiotics and Synbiotics on Apis cerana Gut Microbiota

This study aimed to investigate in vitro effects of the selected prebiotics alone, and in combination with two potential probiotic Lactobacillus strains on the microbial composition of Apis cerana gut microbiota and acid production. Four prebiotics, inulin, fructo-oligosaccharides, xylo-oligosaccharides, and isomalto-oligosaccharides were chosen, and glucose served as the carbon source. Supplementation of this four prebiotics increased numbers of Bifidobacterium and lactic acid bacteria while decreasing the pH value of in vitro fermentation broth inoculated with A. cerana gut microbiota compared to glucose. Then, two potential probiotics derived from A. cerana gut at different dosages, Lactobacillus helveticus KM7 and Limosilactobacillus reuteri LP4 were added with isomalto-oligosaccharides in fermentation broth inoculated with A. cerana gut microbiota, respectively. The most pronounced impact was observed with isomalto-oligosaccharides. Compared to isomalto-oligosaccharides alone, the combination of isomalto-oligosaccharides with both lactobacilli strains induced the growth of Bifidobacterium, LAB, and total bacteria and reduced the proliferation of Enterococcus and fungi. Consistent with these results, the altered metabolic activity was observed as lowered pH in in vitro culture of gut microbiota supplemented with isomalto-oligosaccharides and lactobacilli strains. The symbiotic impact varied with the types and concentration of Lactobacillus strains and fermentation time. The more effective ability was observed with IMO combined with L. helveticus KM7. These results suggested that isomalto-oligosaccharides could be a potential prebiotic and symbiotic with certain lactobacilli strains on A. cerana gut microbiota.     

Mycena kentingensis,a new species of luminous mushroom in Taiwan,with reference to its culture method

Mycena kentingensis, a new luminous mushroom, was discovered in the tropical forest of Kenting National Park in Taiwan. The pileus of M. kentingensis is 3–8 mm in diameter and emits green light in the dark. The dorsal surface of the pileus is covered with short and white spines. The lamellae are nearly free. The stipe is puberulous with a white basal disc about 2 mm broad. Cheilocystidia are clavate to obpyriform, 16–40?×?4–8 μm, and have dense excrescences on the upper half or two-thirds of the cell. The pileus lacks cherocyte, and the stipe lacks caulocystidium. To further identify the species by the DNA barcode method, the internal transcribed spacer (ITS) DNA sequence of M. kentingensis was amplified by polymerase chain reaction, sequenced, and searched for in the GenBank. The hit with the maximum score was an uncultured fungus clone KFRI2121 (accession number HQ662846) with 88 % sequence identity. For those ITS sequences associated with Latin binominal names in the GenBank, the hit with the maximum score was Mycena stylobates with 87 % sequence identity. Phylogenetic analyses by Bayesian and maximum likelihood methods showed that M. kentingensis is close to M. stylobates and M. adscendens. To develop its culture method, mycelia of M. kentingensis were isolated. The optimal temperature for mycelial growth on potato-dextrose agar (PDA) medium was 24 °C. Commercial compost with the addition of 20 % rice bran and 50 % water was appropriate for fruit-body formation. The new species of Mycena kentingensis brings the total of luminous fungi in the world to 74 species.     

Enhancement of the mutagenicities of N-methyl-N′-nitro-N-nitrosoguanidine and N-methyl-N-nitrosourea by glucose

The mutagenicity of N-methyl-N′-nitro-N-nitrosoguanidine to Salmonella typhimurium hisG46 was enhanced by pre-incubating the chemical with bacteria in sodium phosphate buffer. Addition of glucose (to 15 mM) to the pre-incubation mixture further enhanced the mutagenicity. Pre-incubation with glucose also increased the mutagenicity of N-methyl-N-nitrosourea. Fructose, galactose, pyruvate and succinate also enhanced the mutagenicity of N-methyl-N′-nitro-N-nitrosoguanidine. The effect of glucose was observed with S. typhimurium strains hisG46, TA1975, TA1950, TA1535 and TA100.     

Bacterial Diversity and Community Structure in the Pine Wood Nematode Bursaphelenchus xylophilus and B. mucronatus with Different Virulence by High-Throughput Sequencing of the 16S rDNA

Bursaphelenchus xylophilus is the pathogen of pine wilt disease. Bursaphelenchus mucronatus is similar to B. xylophilus in morphology. Both species share a common niche, but they are quite different in pathogenicity. Presently, the role of bacteria in pine wilt disease development has been widely speculated. The diversity of bacteria associated with B. xylophilus and B. mucronatus with different virulence remains unclear. In this study, virulence of four B. xylophilus and four B. mucronatus strains were evaluated by inoculating Pinus thunbergii. High-throughput sequencing targeted 16S rDNA of different virulence nematode strains was carried out. The associated bacterial community structures of the eight strains were analyzed. The results showed that 634,051 high-quality sequences were obtained from the eight nematode strains. The number of OTUs of bacteria associated with B. mucronatus was generally greater than those of B. xylophilus. The richness of the community of bacteria associated with high virulent B. xylophilus ZL1 and AmA3 was higher than moderately virulent B. xylophilus AA3, HE2, and all B. mucronatus strains. While the diversity of bacteria associated with B. mucronatus was higher than B. xylophilus. Stenotrophomonas, Pseudomonadaceae_Unclassified or Rhizobiaceae_Unclassified were predominant in the nematode strains with different virulence. Oxalobacteraceae and Achromobacter were found more abundant in the low virulent B. xylophilus and non-virulent B. mucronatus strains.     

Morphology and anatomy of male cones of Pseudotaxus chienii (W.C. Cheng) W.C. Cheng (Taxaceae)

Results of the present study indicate that male cones of Pseudotaxus chienii are representing inflorescences with strongly reduced flowers. The results fit quite well with investigations showing that sporangiophores of Taxus and also of Pseudotaxus comply with reduced flowers. The only difference between male cones in Taxus and Pseudotaxus is the absence of pherophylls in Taxus. Furthermore our results complete a transition series beginning with Cephalotaxus going on to Pseudotaxus and ending with Taxus and Torreya. In this progression Pseudotaxus can be regarded as an intermediate link between the inflorescences of Cephalotaxus and the simple, unbranched cones of Taxus. The entire transition series shows that sporophyll-like sporangiophores can be derived by reduction of lateral cones. There is however no sign that a similar process has occurred in other conifer groups.     

A multichannel atomic absorption instrument: simultaneous analysis of zinc, copper, and cadmium in biologic materials

A new method is deseribed for the extraction and determination of chlorophylls a and b. The method is suitable for use with both normal and regreening nitrogen-deficient Chlorella fusca. The assay involves extraction of chlorophylls by an alkaline pyridine reagent which converts the isocyclic ring of the pigment to a cyclic hydroxylactone. Millimolar extinetion coefficients for the hydroxylactone derivatives of both chlorophylls a and b have been determined at 419 and 454 nm. Using these coefficients, equations have been derived for the calculation of chlorophyll a and b concentrations. The new chlorophyll assay has been compared with other assays which involve the extraction of the pigments with 80% acetone or methanol. The new procedure extracts chlorophylls from rormally grown C. fusca more readily than methanol; the chlorophylls of normal Chlorella cannot be extracted with 80% acetone. The new assay is especially useful in the study of chlorophyll synthesis in regreening nitrogen-deficient C. fusca since the chlorophylls present in these deficient cells cannot be completely extracted with acetone, methanol, methanol-dimethylsulphoxide mixtures, or KOH-methanol.     

Porphyromonas gingivalis,Aggregatibacter actinomycetemcomitans,and Treponema denticola / Prevotella intermedia Co-Infection Are Associated with Severe Periodontitis in a Thai Population

Periodontitis is a polymicrobial infection of tooth-supporting tissues. This cross-sectional study aimed to examine the associations between five target species and severe periodontitis in a Thai population. Using the CDC/AAP case definition, individuals diagnosed with no/mild and severe periodontitis were included. Quantitative analyses of Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf), Treponema denticola (Td), and Prevotella intermedia (Pi) in subgingival plaque were performed using real-time polymerase chain reaction. The association between target species and severe periodontitis was examined using logistic regression analysis. The study subjects comprised 479 individuals with no/mild periodontitis and 883 with severe periodontitis. Bacterial prevalence and quantity were higher in subjects with severe periodontitis than in those with no/mild disease. In the fully adjusted model, all species except Tf showed a dose-dependent relationship with periodontitis. The mere presence of Pg, even in low amount, was significantly associated with severe periodontitis, while the amount of Aa, Td, and Pi had to reach the critical thresholds to be significantly associated with disease. Compared to individuals with low levels of both Td and Pi, high colonization by either Td or Pi alone significantly increased the odds of having severe periodontitis by 2.5 (95%CI 1.7–3.5) folds. The odds ratio was further increased to 14.8 (95%CI 9.2–23.8) in individuals who were highly colonized by both species. Moreover, the presence of Pg and high colonization by Aa were independently associated with severe periodontitis with odds ratios of 5.6 (95%CI 3.4–9.1) and 2.2 (95%CI 1.5–3.3), respectively. Our findings suggest that the presence of Pg and high colonization by Aa, Td, and Pi play an important role in severe periodontitis in this study population. We also demonstrate for the first time that individuals co-infected with Td and Pi were more likely to have periodontitis than were those infected with a single pathogen.     

Prevalence of Leishmania RNA virus in Leishmania parasites in patients with tegumentary leishmaniasis: A systematic review and meta-analysis

BackgroundCutaneous leishmaniasis is caused by different protozoan parasites of the genus Leishmania. Leishmania RNA virus (LRV) was identified as the first Leishmania infecting virus in 1998. Different studies showed the presence and role of the LRV in Leishmania parasites causing cutaneous leishmaniasis (CL). However, there is limited data on the pooled prevalence of LRV in Leishmania parasites causing CL. Therefore, the aim of this systematic review and meta-analysis was to determine the pooled prevalence of LRV in Leishmania parasite isolates and/or lesion biopsies in patients with CL from the available literature globally.MethodologyWe retrieved the studies from different electronic databases. The studies were screened and identified based on the inclusion and exclusion criteria. We excluded studies exclusively done in experimental animals and in vitro studies. The review was conducted in line with PRISMA guidelines. The meta-analysis was performed with Stata software version 14 with metan command. The forest plot with random-effect model was used to estimate the pooled prevalence with 95% confidence interval. Inverse variance index (I²) was used to assess the heterogeneity among the included articles.Principal findingsA total of 1215 samples from 25 studies were included. Of these, 40.1% (487/1215) were positive for LRV. The overall pooled prevalence of LRV globally was 37.22% (95% CI: 27.54% - 46.90%). The pooled prevalence of LRV in the New World (NW) and Old World (OW) regions was 34.18% and 45.77%, respectively. Leishmania guyanensis, L. braziliensis, L. major, and L. tropica were the most studied species for the detection of LRV. The prevalence of LRV from Leishmania isolates and lesion biopsies was 42.9% (349/813) and 34.3% (138/402), respectively.ConclusionThis systematic study revealed that there is high prevalence of LRV in Leishmania parasites isolated from patients with CL. More comprehensive studies would be required to investigate the presence of the LRV in other Leishmania species such as L. aethiopica to fully understand the role of LRV in different clinical manifestations and disease pathology presented in CL patients.     

PHYLOGENETIC STUDIES OF THE GENUSARTHONIA

A phylogenetic analysis is presented, based on 57 morphological, anatomical, chemical and ecological characters, including 19 species from the generaArthonia,ArthotheliumandSyncesia. The aim of the study was to test the monophyly of some of the groups within the genusArthoniasuggested, for example, by Redinger (e.g. species with reddish ascomata, with grey–pruinose ascomata and with brown–black hypothecia). The results strongly support thatArthoniaandArthotheliumare paraphyletic genera. The best-supported node contains allArthoniaspecies together withA. crozalsianade Lesd.,A. ruanaA. Massal. (both hitherto placed inArthothelium) andSyncesia myrticola(Fée) Tehler. A well-supported clade is formed by a group of pioneer species, often non-lichenized or poorly lichenized, including the type species ofArthonia,A. radiata. The species with reddish and/or K+ reddish ascomata form one clade and the species with more or less brownish or blackish hypothecia form another clade withSyncesia myrticola, the sister group to theOpegraphaceaeandRoccellaceae. The results are discussed and compared with Redinger’s grouping. Relationships to other genera within Arthoniaceae are briefly discussed.Arthothelium scandinavicumTh. Fr.,Arthonia dispersa(Schrad.) Nyl.,A. punctiformisAch. andA. mediellaNyl. are lectotypified.     

Mitochondrial DNA Genomes Organization and Phylogenetic Relationships Analysis of Eight Anemonefishes (Pomacentridae: Amphiprioninae)

Anemonefishes (Pomacentridae Amphiprioninae) are a group of 30 valid coral reef fish species with their phylogenetic relationships still under debate. The eight available mitogenomes of anemonefishes were used to reconstruct the molecular phylogenetic tree; six were obtained from this study (Amphiprion clarkii, A. frenatus, A. percula, A. perideraion, A. polymnus and Premnas biaculeatus) and two from GenBank (A. bicinctus and A. ocellaris). The seven Amphiprion species represent all four subgenera and P. biaculeatus is the only species from Premnas. The eight mitogenomes of anemonefishes encoded 13 protein-coding genes, two rRNA genes, 22 tRNA genes and two main non-coding regions, with the gene arrangement and translation direction basically identical to other typical vertebrate mitogenomes. Among the 13 protein-coding genes, A. ocellaris (AP006017) and A. percula (KJ174497) had the same length in ND5 with 1,866 bp, which were three nucleotides less than the other six anemonefishes. Both structures of ND5, however, could translate to amino acid successfully. Only four mitogenomes had the tandem repeats in D-loop; the tandem repeats were located in downstream after Conserved Sequence Block rather than the upstream and repeated in a simply way. The phylogenetic utility was tested with Bayesian and Maximum Likelihood methods using all 13 protein-coding genes. The results strongly supported that the subfamily Amphiprioninae was monophyletic and P. biaculeatus should be assigned to the genus Amphiprion. Premnas biaculeatus with the percula complex were revealed to be the ancient anemonefish species. The tree forms of ND1, COIII, ND4, Cytb, Cytb+12S rRNA, Cytb+COI and Cytb+COI+12S rRNA were similar to that 13 protein-coding genes, therefore, we suggested that the suitable single mitochondrial gene for phylogenetic analysis of anemonefishes maybe Cytb. Additional mitogenomes of anemonefishes with a combination of nuclear markers will be useful to substantiate these conclusions in future studies.