• Title/Summary/Keyword: Clonal diversity

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Isolation and Genotyping of Toxoplasma gondii in Brazilian Dogs

  • da Silva, Jamille Rodrigues;Maciel, Bianca Mendes;de Santana Souza Santos, Luana Karla Nogueira;Carvalho, Fabio Santos;de Santana Rocha, Daniele;Lopes, Carlos Wilson Gomes;Albuquerque, George Rego
    • Parasites, Hosts and Diseases
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    • v.55 no.3
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    • pp.239-246
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    • 2017
  • Strains of Toxoplasma gondii in Brazil are highly genetically diverse compared to strains from North America and Europe. Dogs are epidemiologically important because they act as sentinels for T. gondii infections in humans and are good indicators of environmental contamination. The aim of this study was to isolate and genetically characterize T. gondii strains from tissues of naturally infected Brazilian dogs. For this study, 21 blood samples were collected from dogs at the Zoonosis Control Centers of $Ilh{\acute{e}}us$ and Itabuna cities, Bahia, Brazil. The sera were examined for T. gondii antibodies using the indirect hemagglutination test. Brains and hearts of seropositive dogs were bioassayed in mice to isolate and characterize T. gondii parasites by PCR-RFLP using 10 genetic markers (SAG1, newSAG2, SAG3, BTUB, c22-8, c29-2, GRA6, PK1, APICO, and L358). However, T. gondii was isolated from only 4 (57.1%) dogs, designated TgDgBr6, 13, 17, and 21. All strains were virulent, causing clinical changes (rough hair coat, lethargy, and abdominal distention) and the death of all mice within 8-20 days after inoculation. Genetic analysis of these 4 T. gondii isolates revealed 4 distinct genotypes with different clonal lineage combinations (types I, II, and III) and 2 atypical alleles. Using PCR-RFLP with several markers, this study contributes to evaluations of the genetic diversity of strains circulating in Brazil.

Genetic Diversity of Toxoplasma gondii Strains from Different Hosts and Geographical Regions by Sequence Analysis of GRA20 Gene

  • Ning, Hong-Rui;Huang, Si-Yang;Wang, Jin-Lei;Xu, Qian-Ming;Zhu, Xing-Quan
    • Parasites, Hosts and Diseases
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    • v.53 no.3
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    • pp.345-348
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    • 2015
  • Toxoplasma gondii is a eukaryotic parasite of the phylum Apicomplexa, which infects all warm-blood animals, including humans. In the present study, we examined sequence variation in dense granule 20 (GRA20) genes among T. gondii isolates collected from different hosts and geographical regions worldwide. The complete GRA20 genes were amplified from 16 T. gondii isolates using PCR, sequence were analyzed, and phylogenetic reconstruction was analyzed by maximum parsimony (MP) and maximum likelihood (ML) methods. The results showed that the complete GRA20 gene sequence was 1,586 bp in length among all the isolates used in this study, and the sequence variations in nucleotides were 0-7.9% among all strains. However, removing the type III strains (CTG, VEG), the sequence variations became very low, only 0-0.7%. These results indicated that the GRA20 sequence in type III was more divergence. Phylogenetic analysis of GRA20 sequences using MP and ML methods can differentiate 2 major clonal lineage types (type I and type III) into their respective clusters, indicating the GRA20 gene may represent a novel genetic marker for intraspecific phylogenetic analyses of T. gondii.

Differential Growth Response of $A_1\;and\;A_2$ Mating Types of Phytophthora infestans on Rye A and V-8 Juice Agar Media Supplemented with Rhizome Powder of Cyperus rotundus

  • Singh, U.P.;Sarma, B.K.;Nishimura, Ruo;Kobayashi, Kiroku;Ogoshi, Akira;Zinkernagel, Volker;Schlenzig, Alexendra;Schober-Butin, Barbel;Aust, H.J.
    • Mycobiology
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    • v.29 no.3
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    • pp.164-169
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    • 2001
  • A new medium for studies of diversity among populations of $A_1\;and\;A_2$ mating types of Phytophthora infestans has been evolved. The rye A agar and V-8 juice agar media on which P. infestans grows well have been amended with rhizome powder of Cyperus rotundus. A total of 259 isolates of $A_1\;and\;A_2$ mating types representing Japan, Korea, India, Taiwan, Indonesia, Thailand, China, Nepal, U.K and Medico were screened for their growth response on these two media. Most of the A1 isolates did not grow well on them except Thailand while growth of $A_2$ mating types differed as some grew on it whereas others did not. It is quite likely that the populations of $A_2$ mating types that did not grow well on rhizome-amended medium are of different clonal lineage. This suggests that this medium can be used for the study of diversification among the isolates of the same or both the mating types as well as to detect the newly introduced genetically different isolates of P. infestans in a locality where it was not reported earlier.

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Molecular Profiling of Rhizosphere Bacterial Communities Associated with Prosopis juliflora and Parthenium hysterophorus

  • Jothibasu, K.;Chinnadurai, C.;Sundaram, S.P.;Kumar, K.;Balachandar, D.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.301-310
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    • 2012
  • Prosopis juliflora and Parthenium hysterophorus are the two arid, exotic weeds of India that are characterized by distinct, profuse growth even in nutritionally poor soils and environmentally stressed conditions. Owing to the exceptional growth nature of these two plants, they are believed to harbor some novel bacterial communities with wide adaptability in their rhizosphere. Hence, in the present study, the bacterial communities associated with the rhizosphere of Prosopis and Parthenium were characterized by clonal 16S rRNA gene sequence analysis. The culturable microbial counts in the rhizosphere of these two plants were higher than bulk soils, possibly influenced by the root exudates of these two plants. The phylogenetic analysis of V1_V2 domains of the 16S rRNA gene indicated a wider range of bacterial communities present in the rhizosphere of these two plants than in bulk soils and the predominant genera included Acidobacteria, Gammaproteobacteria, and Bacteriodetes in the rhizosphere of Prosopis, and Acidobacteria, Betaproteobacteria, and Nitrospirae in the Parthenium rhizosphere. The diversity of bacterial communities was more pronounced in the Parthenium rhizosphere than in the Prosopis rhizosphere. This culture-independent bacterial analysis offered extensive possibilities of unraveling novel microbes in the rhizospheres of Prosopis and Parthenium with genes for diverse functions, which could be exploited for nutrient transformation and stress tolerance in cultivated crops.

Characteristics of Transmissible CTX-M- and CMY-Type β-Lactamase-Producing Escherichia coli Isolates Collected from Pig and Chicken Farms in South Korea

  • Shin, Seung Won;Jung, Myunghwan;Won, Ho Geun;Belaynehe, Kuastros Mekonnen;Yoon, In Joong;Yoo, Han Sang
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1716-1723
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    • 2017
  • The rapid dissemination of extended-spectrum ${\beta}$-lactamase (ESBL)-producing Escherichia coli has significantly contributed to public health hazard globally. A total of 281 E. coli strains recovered from pigs and chickens between 2009 and 2015 in South Korea were analyzed for ESBL production. ESBL phenotypes were recognized in 14 E. coli isolates; ten and three ESBL-producing isolates carried only $bla_{CTX-M}$ and $bla_{CMY}$ genes, respectively, and one isolate harbored both genes. The predominant CTX-M and CMY types were CTX-M-15 (n = 8) and CMY-2 (n = 3). We also detected ESBL-producing isolates harboring $bla_{CTX-M-65}$, $bla_{CTX-M-14}$, $bla_{CMY-6}$, $bla_{DHA-1}$, and $bla_{TEM-1}$ genes. All ESBL-producing isolates showed resistance to the extent of the fourth generation cephalosporins, along with multidrug resistance. CTX-M-15-producing isolates showed higher MIC values than CTX-M-14- and CTX-M-65-producing isolates. The $bla_{CTX-M}$ and $bla_{CMY}$ genes have the potential to be transferable. The spreading of $bla_{CMY}$ and $bla_{CTX-M}$ genes was arbitrated mainly via Frep and IncI1 plasmids. Our isolates showed clonal diversity in PFGE analysis. This is the first report of E. coli isolates carrying $bla_{CMY-6}$ in chicken from South Korea. The emergence of CMY-6 ESBLs in a population of poultry suggests that extensive screening with long-term surveillance is necessary to prevent the dissemination of ESBL from chicken to human.

Serotype Distribution and Virulence Profile of Salmonella enterica Serovars Isolated from Food Animals and Humans in Lagos Nigeria

  • Abraham, Ajayi;Stella, Smith;Ibidunni, Bode-Sojobi;Coulibaly, Kalpy Julien;Funbi, Jolaiya Tolulope;Isaac, Adeleye Adeyemi
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.310-316
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    • 2019
  • Distribution of Salmonella enterica serovars and their associated virulence determinants is wide-spread among food animals, which are continuously implicated in periodic salmonellosis outbreaks globally. The aim of this study was to determine and evaluate the diversity of five Salmonella serovar virulence genes (invA, pefA, cdtB, spvC and iroN) isolated from food animals and humans. Using standard microbiological techniques, Salmonella spp. were isolated from the feces of humans and three major food animals. Virulence determinants of the isolates were assayed using PCR. Clonal relatedness of the dominant serovar was determined via pulsed-field gel electrophoresis (PFGE) using the restriction enzyme, Xbal. Seventy one Salmonella spp. were isolated and serotyped into 44 serovars. Non-typhoidal Salmonella (NTS; 68) accounted for majority (95.8%) of the Salmonella serovars. Isolates from chicken (34) accounted for 47.9% of all isolates, out of which S. Budapest (14) was predominant (34.8%). However, the dominant S. Budapest serovars showed no genetic relatedness. The invA gene located on SPI-1 was detected in all isolates. Furthermore, 94% of the isolates from sheep harbored the spvC genes. The iroN gene was present in 50%, 100%, 88%, and 91% of isolates from human, chicken, sheep, and cattle, respectively. The pefA gene was detected in 18 isolates from chicken and a single isolate from sheep. Notably, having diverse Salmonella serovars containing plasmid encoded virulence genes circulating the food chain is of public health significance; hence, surveillance is required.

Species Diversity of the Dinoflagellate Genus Alexandrium in the Coastal Waters of Korea during Summer 2013 (2013년 하계 국내 연안에서 출현하는 Alexandrium 속 와편모류의 종 다양성)

  • KIM, JAE SEONG;PARK, KYUNG WOO;YOUN, SEOK HYUN;LIM, WEOL AE;YOO, YEONG DU;SEONG, KEYONG AH;YIH, WON HO
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.21 no.4
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    • pp.158-170
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    • 2016
  • We investigated the occurrence of the dinoflagellate genus Alexandrium in the nineteen Korean coastal sites from July to October 2013. Alexandrium-like planktonic cells were microscopically observed only in four out of the 19 sampling sites. From the samples containing Alexandrium-like cells 22 clonal cultures of Alexandrium species were established by single cell or single chain isolation method. Taxonomic identity of the 4 different strains ascertained by the robust analyses of morphological and molecular genetic characteristics were confirmed to be A. catenella, A. affine, A. fraterculus and an unidentified Alexandrium sp. for which strain WEB-Alex-01 was assigned. It was ascertained that in spite of hot summer diverse Alexandrium species attaining up to four were distributed in the study area, in contrast with the long empirical recognition that the emergence of Alexandrium species is restricted to cooler seasons like spring or autumn in Korean coastal waters. Morphology and genetic characteristics of Alexandrium sp. strain WEB-Alex-01 are different from any other previously reported Alexandrium species from Korean seas, which implies that further studies on taxonomic, physiological, ecological and toxicological properties of the newly recorded Alexandrium species are needed.

Characteristics of Plant Community of Willow Forest in the Wetland Protection Areas of Inland Wetlands (습지보호지역 버드나무림의 식물군락 특성)

  • Cho, Kwang-Jin;Lim, Jeoncheol;Lee, Changsu;Chu, Yeounsu
    • Journal of Wetlands Research
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    • v.23 no.3
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    • pp.201-212
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    • 2021
  • In wetland protection areas, a phytosociological research was conducted on willow forests, which plays an important ecological and environmental role and is the main material for ecological restoration. A total of 61 relevés were collected according to the Z-M(Zürich-Montpellier) school's method and the characteristics of plant communities and the composition of the species were identified. A total of 9 plant communities including 237 taxa were differentiated. Willow species showing the high r-NCD(relative net contribution degree) value in study areas were Salix koriyanagi, Salix chaenomeloides, Salix triandra subsp. nipponica, Salix gracilistyla and Salix pierotii. Poaceae was the most diverse in species, followed by Asteraceae, Apiaceae, Polygonaceae and Fabaceae. Life form type of willow forests in wetland protection areas was determined to be Th(therophytes)-R5(non-clonal form)-D4(clitochores)-e(erect form) type. The naturalized plants and invasive alien species were identified as 24 taxa and 4 taxa, respectively. Naturalized and disturbance indices were 10.1% and 41.4%, respectively. The results of the site-species ordination by Non-Metric Multidimensional Scaling(NMDS), wetland types and altitude gradient were the main ecological factors determining the spatial distribution of plant communities. Diversity index and evenness index were high in mountainous palustrine wetlands with relatively high altitude, and the disturbance index, naturalized index and appearance rate of annual plant were high in riverine and lacustrine wetlands with low altitude.