• Title/Summary/Keyword: small subunit rRNA

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Novel Discovery of Two Heterotrichid Ciliates, Climacostomum virens and Fabrea salina (Ciliophora: Heterotrichea: Heterotrichida) in Korea

  • Kim, Ji Hye;Shin, Mann Kyoon
    • Animal Systematics, Evolution and Diversity
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    • v.31 no.3
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    • pp.182-190
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    • 2015
  • Two heterotrichid ciliates, Climacostomum virens (Ehrenberg, 1838) Stein, 1859 from brackish water and freshwater, and Fabrea salina Henneguy, 1890 from a solar saltern, were collected in Korea. They are novelly investigated in Korea by means of live observation, protargol staining and nuclear small subunit (SSU) rRNA gene sequencing. Climacostomum virens is characterized by pouch-like body shape, body length of $200-370{\mu}m$ in vivo, conspicuous cytopharyngeal tube, macronuclei ribbon-like shape, and one to four in number, with or without symbiont algae in cytoplasm, 34-66 somatic kineties, 67-113 adoral zone of membranelles, 8-42 peristomial kineties, 24-37 apical membranelles. SSU rDNA sequence size is 1,591 bp and GC contents 48.52%. Fabrea salina is also characterized by scoop-like body shape with proboscis, body length of $190-240{\mu}m$ in vivo, one to two rod-shaped macronuclei, oval micronuclei, grayish green cortical granules, 104-186 somatic kineties, 4-8 preoral kineties, 7-19 peristomial kineties and fragmented paroral membrane. SSU rDNA sequence size is 1,598 bp and GC contents 47.50%.

Molecular Characterization of Various Trichomonad Species Isolated from Humans and Related Mammals in Indonesia

  • Kamaruddin, Mudyawati;Tokoro, Masaharu;Rahman, Md. Moshiur;Arayama, Shunsuke;Hidayati, Anggi P.N.;Syafruddin, Din;Asih, Puji B.S.;Yoshikawa, Hisao;Kawahara, Ei
    • Parasites, Hosts and Diseases
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    • v.52 no.5
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    • pp.471-478
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    • 2014
  • Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

A case report of farmed olive flounder Paralichthys olivaceus infected with Myxosporean Parvicapsula anisocaudata (양식 넙치에서 Parvicapsula anisocaudata의 감염 사례에 대한 보고)

  • Kim, Nam Eun;Kim, Ahran;Roh, Heyong Jin;Gang, Kyoung Sik;Kim, Do-Hyung
    • Journal of fish pathology
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    • v.31 no.2
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    • pp.123-129
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    • 2018
  • Parvicapsula anisocaudata, a myxosporean parasite, is presumably one of causative agents of emaciation in olive flounder Paralichthys olivaceus in Korea. In this study, we report a case of unusual abdominal distension due to exceptionally enlarged liver in farmed olive flounder. For the identification of the causative agent, bacteria and nucleic acids of virus that are possibly present were attempted to isolate from internal organs of five fish sampled from a fish farm in Jeju. Although a few bacterial colonies were isolated from some samples, there was no evidence that fish were primarily affected by virus and/or bacteria. From histopathological analysis, myxosporean were found in almost all internal organs, particularly in the stomach. The causative agent was identified as P. anisocaudata by sequencing a part of small subunit rRNA. This study contains a very unusual case of olive flounder heavily and systemically infected with P. anisocaudata, showing excessively enlarged liver with a small amount of ascitic fluid.

Morphological and Molecular Identification of Pseudo-nitzschia sp. Strain G3 Isolated from Northern Coast of Vietnam Based on ITS Region Sequences

  • Dang, Diem-Hong;Luyen, Hai-Quoc;Hien, Hoang Thi Minh;Thu, Ngo Hoai;Anh, Hoang Lan
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.1
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    • pp.60-67
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    • 2007
  • For the first time in Vietnam, morphological and molecular studies of a species belonging to Bacillariophyceae collected in Northern coast of Vietnam are presented. Observations with microscope showed that this species belong to genus: Pseudo-nitzschia and seem like P. pungens. Sequence data from the partial 18S small subunit ribosomal RNA gene (18S rDNA) and the internal transcribed spacer 1 - 5.8S - internal transcribed 2 have been used to determine clearly and generate a phylogenetic framework of the obtained sequences to previously reported sequences in GenBank. These results allowed us to highlight described species of Bacillariophyceae in Northern coast of Vietnam. Furthermore, accumulation of molecular study would be helpful for the identification of scientific name of harmful algal species and further taxonomic studies in Vietnam.

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PCR and RFLP variation of conserved region of small subunit ribosomal DNA among Acanthamoeba isolates assigned to either A. castellanii or A. polyphaga (카스텔라니가시아메바 혹은 대식가시아메바로 분류된 분리주간의 ribosomaIDNA conserved region의 PCR-RFLP의 다양성)

  • 공현희;정동일
    • Parasites, Hosts and Diseases
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    • v.34 no.2
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    • pp.127-134
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    • 1996
  • Twelve isolates of Accnthamoebc app. assigned to either A. castellanii or A. poIMphoSa, and type strains of A. culbefsoni, A. henIWi, A. pqkestinefiE, and A. astronyxi,s were examined by restriction fragment length polymorphism (RFLP) of a conserved region of small subunit ribosomal RMh gene (ssu rDNA) amplified by polymerase chain reaction (PCR). The PCR products of the isolates measured approximately 910-930 bp, except for that of A. astronyxis which was extraordinarily long, approximately 1,170 bp. Average of estimated sequence divergence of the amplified DNA among the isolates assigned to A. castellanii was 9.8% whereas that among the isolates assigned to A. polvphusn 9.6%. The maximum intraspecific sequence divergence among the isolates assigned to A. costellanii was observed between the Chang and Ma strains (17.3%) while that among the isolates assigned to A. poIWphosa was observed between KA/S3 and KA/S7 strains (16.1%). The both maximum sequence divergences were much greater than the minimum interspecific sequence divergence between A. cnstellnnii and A. polwphasa (2.6%) which appeared between the Castellani (or CCAP 1501/12 g) and KA/S3 strains. The PCR-RFLP patterns of A. culbertsoni, A. healyi, A. palestinensis, and A. ostronvxis were quite diverse from one another and from those of isolates assigned to either A. castellanii or A. polyphoga. It is suggested that taxonomic validity of the isolates assigned to either A. castellnnii or A. polyphoga should be reevaluated.

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Molecular Prevalence of Cryptosporidium spp. among Companion Birds Kept in Pet Shops in Japan

  • Iijima, Yuko;Itoh, Naoyuki;Phrompraphai, Totsapon;Ito, Yoichi;Kimura, Yuya;Kameshima, Satoshi
    • Parasites, Hosts and Diseases
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    • v.56 no.3
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    • pp.281-285
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    • 2018
  • Cryptosporidium is the most common protozoan that can infect a wide range of animals, including mammals and birds. Avian Cryptosporidium spp. can cause enteric and respiratory diseases which can be fatal in birds and some species are zoonotic. Companion birds have the potential as reservoir due to their close contact with humans. Pet shops are the major source of companion birds. However, few reports are available regarding Cryptosporidium spp. infection among companion birds kept in pet shops. The present study reports the prevalence and molecular characteristics of Cryptosporidium spp. among companion birds kept in pet shops in Japan. A total of 265 fresh fecal samples were obtained from birds kept in 4 pet shops; these birds belonged to 41 species in 3 bird orders. A nested polymerase chain reaction (PCR) assay targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. A total of 24 samples (9.1%) were positive, and Cryptosporidium spp. were detected from all pet shops. The prevalence of Cryptosporidium spp. in each of the bird orders was 6.5% (10/153) in Psittaciformes, 14.4% (13/90) in Passeriformes, and 4.5% (1/22) in Galliformes. Based on sequence analysis, 13 (54.2%) isolates were classified to C. galli, 8 (33.3%) were avian genotype III, and the remaining 3 (12.5%) were C. baileyi. No infection with zoonotic C. meleagridis and no coinfection with multiple Cryptosporidium spp. and/or genotypes were observed. The zoonotic potential of Cryptosporidium spp. infecting companion birds kept in pet shops in Japan is likely to be low.

Complete Mitochondrial Genome Sequences of Chinese Indigenous Sheep with Different Tail Types and an Analysis of Phylogenetic Evolution in Domestic Sheep

  • Fan, Hongying;Zhao, Fuping;Zhu, Caiye;Li, Fadi;Liu, Jidong;Zhang, Li;Wei, Caihong;Du, Lixin
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.5
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    • pp.631-639
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    • 2016
  • China has a long history of sheep (Ovis aries [O. aries]) breeding and an abundance of sheep genetic resources. Knowledge of the complete O. aries mitogenome should facilitate the study of the evolutionary history of the species. Therefore, the complete mitogenome of O. aries was sequenced and annotated. In order to characterize the mitogenomes of 3 Chinese sheep breeds (Altay sheep [AL], Shandong large-tailed sheep [SD], and small-tailed Hulun Buir sheep [sHL]), 19 sets of primers were employed to amplify contiguous, overlapping segments of the complete mitochondrial DNA (mtDNA) sequence of each breed. The sizes of the complete mitochondrial genomes of the sHL, AL, and SD breeds were 16,617 bp, 16,613 bp, and 16,613 bp, respectively. The mitochondrial genomes were deposited in the GenBank database with accession numbers KP702285 (AL sheep), KP981378 (SD sheep), and KP981380 (sHL sheep) respectively. The organization of the 3 analyzed sheep mitochondrial genomes was similar, with each consisting of 22 tRNA genes, 2 rRNA genes (12S rRNA and 16S rRNA), 13 protein-coding genes, and 1 control region (D-loop). The NADH dehydrogenase subunit 6 (ND6) and 8 tRNA genes were encoded on the light strand, whereas the rest of the mitochondrial genes were encoded on the heavy strand. The nucleotide skewness of the coding strands of the 3 analyzed mitogenomes was biased toward A and T. We constructed a phylogenetic tree using the complete mitogenomes of each type of sheep to allow us to understand the genetic relationships between Chinese breeds of O. aries and those developed and utilized in other countries. Our findings provide important information regarding the O. aries mitogenome and the evolutionary history of O. aries inside and outside China. In addition, our results provide a foundation for further exploration of the taxonomic status of O. aries.

Molecular Prevalence of Cryptosporidium spp. in Breeding Kennel Dogs

  • Itoh, Naoyuki;Tanaka, Hazuki;Iijima, Yuko;Kameshima, Satoshi;Kimura, Yuya
    • Parasites, Hosts and Diseases
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    • v.57 no.2
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    • pp.197-200
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    • 2019
  • Cryptosporidium is a common intestinal protozoan that can lead to diarrhea in humans and dogs. The predominant species of infection are C. hominis and C. parvum in humans, and C. canis in dogs. However, C. canis can infect immunocompromised humans. Considering the close contact with humans, dogs have the potential to be reservoirs for human cryptosporidiosis. Breeding kennels are the major supply source of puppies for pet shops. The present study is to determine the molecular prevalence and characteristics of Cryptosporidium spp. found in breeding kennel dogs. A total of 314 fecal samples were collected from young and adult dogs kept in 5 breeding kennels. A polymerase chain reaction targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. To determine the species, the DNA sequences were compared to GenBank data. Overall, 21.0% of the fecal samples were positive for Cryptosporidium spp. infection. Cryptosporidium spp. was detected in all 5 facilities. A sequencing analysis demonstrated that all isolates shared 99-100% similarity with C. canis. The results suggest that Cryptosporidium spp. infection is present at a high-level in breeding kennel dogs. However, because dominant species in this survey was C. canis, the importance of breeding kennel dogs as reservoirs for Cryptosporidium spp. transmission to humans is likely to be low in Japan.

Cold-Seep Sediment Harbors Phylogenetically Diverse Uncultured Bacteria

  • Cho, Jae-Chang;Lee, Sang-Hoon;Oh, Hae-Ryun;Lee, Jung-Hyun;Kim, Sang-Jin
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.906-913
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    • 2004
  • A culture-independent molecular phylogenetic survey was carried out on the bacterial community in cold-seep sediment at Edison Seamount, south of Lihir Island, Papua New Guinea. Small-subunit rRNA genes were amplified directly from the sediment DNA by PCR and cloned. The majority of the cloned 16S rRNA gene sequences were most closely related to as-yet-uncultivated microorganisms found in deep-sea sediments, and were primarily affiliated with one of four groups: the $\gamma$-, $\delta$-, and $\epsilon$-subdivisions of Proteobacteria, and Cytophaga-Flavobacterium-Bacteroides. We did not recover any sequences related to cyanobacteria, prochlorophytes, and $\alpha$-Proteobacteria, which are known to occur in great abundance within the surface mixed layer of the Atlantic and Pacific Oceans. The majority of the cloned $\gamma$-and $\epsilon$-Proteobacterial sequences were closely related to chemoautotrophic sulfur-oxidizing symbionts of marine benthic fauna, and the $\delta$-Proteobacterial sequences to sulfate- and sulfur-reducing bacteria, indicating that they might play an important role in chemoautotrophic primary production and the sulfur cycle in the cold-seep area. There results demonstrate the high diversity of the bacterial community in the cold-seep sediment, and substantially expand knowledge of the extent of bacterial diversity in this formidable and unique habitat.

Molecular Phylogenetic Relationships Within the Genus Alexandrium(Dinophyceae) Based on the Nuclear-Encoded SSU and LSU rDNA D1-D2 Sequences

  • Kim, Choong-Jae;Sako Yoshihiko;Uchida Aritsune;Kim, Chang-Hoon
    • Journal of the korean society of oceanography
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    • v.39 no.3
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    • pp.172-185
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    • 2004
  • LSU rDNA D1-D2 and SSU rDNA genes of 23 strains in seven Alexandrium (Halim) species, A. tamarense (Lebour) Balech, A. catenella (Whedon et Kofoid), A. fraterculus (Balech) Balech, A. affine (Inoue et Fukuyo) Balech, A. insuetum Balech, A. pseudogonyaulax (Biecheler) Horiguchi ex Yuki et Fukuyo and A. tamiyavanichii Balech, were sequenced and the data were used for molecular phylogenetic analysis. The sequence data revealed 11 and 7 ribotypes in the LSU rDNA D1-D2 region and 4 and 17 ribotypes in the SSU rDNA region of A. catenella and A. tamarense, respectively. Other Alexandrium species had also 1 to 5 ribotypes in the two regions. With the exception of CMC2 and CMC3 of A. catenella, all A. tamarense and A. catenella strains had a common ribotype, a functionally expressed rRNA gene (here termed type A), in both gene regions. In addition to the functionally expressed gene, several pseudogenes were obtained that were found to be good tools to analyze the population designation of regional isolates by grouping them according to shared ribotypes. From the phylogenetic analysis of the sequence data determined in this study and retrieved from GenBank, the genus Alexandrium was divided into 14 groups: 1) A. tamarense, 2) A. excavatum, 3) A. catenella, 4) Tasmanian A. tamarense, 5) A. affine (and/or A. concavum), 6) Thai A. tamarense, 7) A. tamiyavanichii, 8) A. fraterculus, 9) A. margalefii, 10) A. andersonii, 11) A. ostenfeldii, 12) A. minutum (or A. lusitanicum), 13) A. insuetum, and 14) A. pseudogonyaulax. The SSU rDNA gene sequence of A. fundyense was so similar to those of A. tamarense used in this study that the two species were difficult to discriminate each other. A. tamiyavanichii was closest to the A. tamarense strain isolated in Thailand and close to the long chain-forming species of A. affine and A. fraterculus. The phylogenetic tree showed that A. margalefii, A. andersonii, A. ostenfeldii, A. minutum and A. insuetum constituted the basal relative complex, and that A. pseudogonyaulax is an ancestral taxon in the genus Alexandrium.