• 제목/요약/키워드: Species-specific PCR

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Tracing the origin of fish without hatchery information: genetic management of stock enhancement for mangrove red snapper (Lutjanus argentimaculatus) in Taiwan

  • Hsu, Te-Hua;Huang, Chang-Wen;Lin, Cheng-Hui;Lee, Hung-Tai;Pan, Chieh-Yu
    • Fisheries and Aquatic Sciences
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    • 제23권5호
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    • pp.13.1-13.7
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    • 2020
  • Stock enhancement is considered to be a valuable approach for restoring fishery resources. Because no specific official institution in Taiwan is responsible for the production of fry, the released fry are purchased directly from the private sector. However, fishermen from the private industry have not established a genetic background, so the genetic composition for each batch of released fry is unclear. Mangrove red snapper (Lutjanus argentimaculatus), a prominent species released in Taiwan, was collected after its official release. One hundred and two field samples were compared with four batches of hatchery fry (n = 685) by using a microsatellite-based multiplex PCR assay. Four of the field samples (3.9%; 4/102) were revealed to be from a fish farm and most likely from a single batch. This study revealed that wild mangrove red snappers are genetically different from those originating from farms, and their origins can be traced through molecular markers, even without information on breeding stocks.

Identification of Two Fungal Endophytes Associated with the Endangered Orchid Orchis militaris L.

  • Vendramin, Elena;Gastaldo, Andrea;Tondello, Alessandra;Baldan, Barbara;Villani, Mariacristina;Squartini, Andrea
    • Journal of Microbiology and Biotechnology
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    • 제20권3호
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    • pp.630-636
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    • 2010
  • A survey of the endangered orchid Orchis militaris populations was carried out in north-eastern Italy. The occurrence of fungal root endophytes was investigated by light and electron microscopies and molecular techniques. Two main sites of presence were individuated in the Euganean Hills, differing as to the percentage of flowering individuals and of capsules completing maturity. Fluorescence microscopy revealed an intracellular cortical colonization by hyphal pelotons. Two ITS PCR products co-amplified. Sequencing revealed for the former an identity and a high similarity (99%) with a Tulasnellaceae (Basidiomycota) fungus found within tissues of the same host in independent studies in Hungary and Estonia, suggesting an interesting case of tight specificity throughout the Eurosiberian home range. The second amplicon had 99% similarity with Tetracladium species (Ascomycota) recently demonstrated as potential endophytes. TEM revealed two different hyphal structures. Double fungal colonization appears to occur in Orchis militaris and the possible requirement of a specific fungal partner throws light on the causes of this plant's rarity and threatened status.

경북지역 돼지 유래 Pasteurella multocida 의 특성 및 항생제 내성양상 (Characteristics and antimicrobial resistance patterns of Pasteurella multocida isolated from swine in Gyeongbuk province)

  • 손준형;김영환;신성호;이은미;김순태;조민희;윤문조
    • 한국동물위생학회지
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    • 제37권3호
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    • pp.165-171
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    • 2014
  • This study was conducted to investigate the species-specific gene detection, capsular serogroup and antimicrobial resistance pattern of Pasteurella multocida isolated from pneumonic lung lesion of swine in Gyeongbuk province. P. multocida isolates were typed for capsular serogroups by polymerase chain reaction. Of the 32 strains, 28 (87.5%) were typed serotype A, 3 (9.3%) were typed serotype D, and 1 strain was unknown (3.1%), respectively. In antimicrobial agents resistance test, almost of strains were susceptible to amoxicillin (100%), enrofloxacin (96.9%), ampicillin (93.8%), florfenicol (90.6%), chloramphenicol (90.6%) and were resistant to streptomycin (71.9%), spectinomycin (56.3%). All strains were resistant to clindamycin, erythromycin and lincomycin.

A Detection Kit for Aeromonas hydrophila Using Antibody Sensitized Latex

  • Shin, En-Joo;Lee, Soon-Deuk;Lee, Kyung-Won;Lee, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • 제10권5호
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    • pp.595-598
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    • 2000
  • Aeromonas hydrophila is a pathogen to fish as well as human. It is a food-borne disease, and causes severe mortality in fish, and sometimes severe septicemia in human. In this study, a rapid detection method using latex agglutination has been developed for A. hydrophila. Polyclonal antibodies were raised against membrane and whole cells of three isolates from rainbow trout. Among these, latex particles coated with antibodies raised against whole cells of isolate No. 2 showed the best sensitivity. With latex particles coated with this antibody, we could detect $5{\times}10^4$ CFU of A. hydrophila in 5 min. The cross-reactivity with bacteria constituting the normal intestinal microflora and other pathogens for rainbow trout was insignificant. This latex agglutination assay method produced positive reaction with all clinical isolates of A. hydrophila which were identified by species-specific PCR for 16S rRNA in A. hydrophila.

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An Efficient Approach for Cloning P450 Hydroxylase Genes from Actinomycetes

  • Hyun, Chang-Gu;Kim, Jung-Mee;Hong, Soon-Kwang;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • 제8권3호
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    • pp.295-299
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    • 1998
  • Oligonucleotide primers were designed and successfully applied to amplify DNA fragments of P450 hydroxylase genes from actinomycetes which produce a large variety of medically important metabolites. Primers were designed based on several regions of strong similarities in amino acid sequence of P450 hydroxylases from a variety of actinomycetes, primarily in the regions of an oxygen binding site and a heme ligand pocket. These primers were used to amplify DNA fragments from seven different actinomycetes species producing a variety of different compounds. The deduced amino acid sequences of the isolated fragments revealed significant similarities to known P450 hydroxylase including the product of the suaC or subC genes from Streptomyces griseolus that is capable of metabolizing a number of sulfonylurea herbicides, and to the product of the $P450_{sca2}$ from S. carbophilus that produces a specific HMG-CoA reductase inhibitor. This method should help researchers in cloning the P450 hydroxylase genes involved in the biosynthesis of useful compounds.

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돌돔 ferritin H 유전자의 클로닝과 발현 분석 (Molecular cloning and expression analysis of a ferritin H subunit from rock bream, Oplegnathus fasciatus)

  • 권문경;정지민;김주원;박찬일
    • 한국어병학회지
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    • 제26권3호
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    • pp.295-301
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    • 2013
  • Ferritin is an evolutionarily conserved protein that plays an important role in iron storage and detoxification. In this study, the gene encoding a ferritin H subunit homologue (RbFH) was cloned from rock bream (Oplegnathus fasciatus) and analyzed at the expression. The full-length ferritin H cDNA was 1162 bp long and contained an open reading frame (ORF) of 531 bp that encoded 177 amino acid residues with a predicted molecular mass of 20.8 kDa. The 5' UTR was 297 bp in length, and the 3' UTR 298 bp, and preceded by a 5'-untranslated region that contains a putative Iron Regulatory Element (IRE). The deduced amino acid sequence of RbFH shares extensive sequence identities with the H ferritins of a number of fish species and contains the ferroxidase center that is preserved in ferritin H subunits. Examination of tissue specific expression indicated that RbFH expression was most abundant in PBLs, RBC, liver and muscle.

Serological evidence of West Nile viral infection in archived swine serum samples from Peninsular Malaysia

  • Mohammed, Mohammed Nma;Yasmin, Abd Rahaman;Noraniza, Mohd Adzahan;Ramanoon, Siti Zubaidah;Arshad, Siti Suri;Bande, Faruku;Mohammed, Hussni O.
    • Journal of Veterinary Science
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    • 제22권3호
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    • pp.29.1-29.6
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    • 2021
  • West Nile virus (WNV), a neurotropic arbovirus, has been detected in mosquitos, birds, wildlife, horses, and humans in Malaysia, but limited information is available on WNV infection in Malaysian pigs. We tested 80 archived swine serum samples for the presence of WNV antibody and West Nile (WN) viral RNA using ID Screen West Nile Competition Multi-species enzyme-linked immunosorbent assay kits and WNV-specific primers in reverse transcription polymerase chain reaction assays, respectively. A WNV seroprevalence of 62.5% (50/80) at 95% confidence interval (51.6%-72.3%) was recorded, with a significantly higher seroprevalence among young pigs (weaner and grower) and pigs from south Malaysia. One sample was positive for Japanese encephalitis virus antibodies; WN viral RNA was not detected in any of the serum samples.

Microbial population dynamics in constructed wetlands: Review of recent advancements for wastewater treatment

  • Rajan, Rajitha J.;Sudarsan, J.S.;Nithiyanantham, S.
    • Environmental Engineering Research
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    • 제24권2호
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    • pp.181-190
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    • 2019
  • Constructed wetlands are improvised man-made systems, designed for adopting the principle of natural wetlands for purifying wastewater - the elixir of life. They are used widely as a cost-effective and energy-efficient solution for treating greywater generated from different tertiary treatment sources. It provides an elaborate platform for research activities in an attempt to recycle earth's natural resources. Among the several organic impurities removal mechanisms existing in constructed wetland systems, the earth's active microbial population plays a vital role. This review deals with the recent advancements in constructed wetland systems from a microbiological perspective to (effect/ devise/ formulate) chemical and physical treatment for water impurities. It focuses on microbial diversity studies in constructed wetlands, influence of wetland media on microbial diversity and wetland performance, role of specific microbes in water reuse, removal of trace elements, some heavy metals and antibiotics in constructed wetlands. The impurities removal processes in constructed wetlands is achieved by combined interactive systems such as selected plant species, nature of substrate used for microbial diversity and several biogeochemical effected reaction cycles in wetland systems. Therefore, the correlation studies that have been conducted by earlier researchers in microbial diversity in wetlands are addressed herewith.

Lucerne transient streak virus; a Recently Detected Virus Infecting Alfafa (Medicago sativa) in Central Saudi Arabia

  • Raza, Ahmed;Al-Shahwan, Ibrahim M.;Abdalla, Omer A.;Al-Saleh, Mohammed A.;Amer, Mahmoud A.
    • The Plant Pathology Journal
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    • 제33권1호
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    • pp.43-52
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    • 2017
  • A survey was conducted to determine the status of Lucerne transient streak virus (LTSV) in three high-yielding alfalfa regions in central Saudi Arabia (Riyadh, Qassim, and Hail) during 2014. Three hundred and eight symptomatic alfalfa, and seven Sonchus oleraceus samples were collected. DAS-ELISA indicated that 59 of these samples were positive to LTSV. Two isolates of LTSV from each region were selected for molecular studies. RT-PCR confirmed the presence of LTSV in the selected samples using a specific primer pair. Percentage identity and homology tree comparisons revealed that all Saudi isolates were more closely related to each other but also closely related to the Canadian isolate-JQ782213 (97.1-97.6%) and the New Zealand isolate-U31286 (95.8-97.1%). Comparing Saudi isolates of LTSV with ten other sobemoviruses based on the coat protein gene sequences confirmed the distant relationship between them. Eleven out of fourteen plant species used in host range study were positive to LTSV. This is the first time to document that Trifolium alexandrinum, Nicotiana occidentalis, Chenopodium glaucum, and Lathyrus sativus are new host plant species for LTSV and that N. occidentalis being a good propagative host for it.