scholarly journals Co-Isolation and Characterization of Two Pandoraviruses and a Mimivirus from a Riverbank in Japan

Viruses ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 1123 ◽  
Author(s):  
Motohiro Akashi ◽  
Masaharu Takemura
Keyword(s):  
Phylogenetic Analysis ◽  
Dna Analysis ◽  
Virus Isolation ◽  
Random Amplified Polymorphic Dna ◽  
Isolation And Characterization ◽  
The Family ◽  
Complete Sequences ◽  
Infected Amoeba ◽  
Giant Viruses

Giant viruses, like pandoraviruses and mimiviruses, have been discovered from diverse environments, and their broad global distribution has been established. Here, we report two new isolates of Pandoravirus spp. and one Mimivirus sp., named Pandoravirus hades, Pandoravirus persephone, and Mimivirus sp. isolate styx, co-isolated from riverbank soil in Japan. We obtained nearly complete sequences of the family B DNA polymerase gene (polB) of P. hades and P. persephone; the former carried two known intein regions, while the latter had only one. Phylogenetic analysis revealed that the two new pandoravirus isolates are closely related to Pandoravirus dulcis. Furthermore, random amplified polymorphic DNA analysis revealed that P. hades and P. persephone might harbor different genome structures. Based on phylogenetic analysis of the partial polB sequence, Mimivirus sp. isolate styx belongs to mimivirus lineage A. DNA staining suggested that the Pandoravirus spp. asynchronously replicates in amoeba cells while Mimivirus sp. replicates synchronously. We also observed that P. persephone- or Mimivirus sp. isolate styx-infected amoeba cytoplasm is extruded by the cells. To the best of our knowledge, we are the first to report the isolation of pandoraviruses in Asia. In addition, our results emphasize the importance of virus isolation from soil to reveal the ecology of giant viruses.

scholarly journals Random Amplification of Polymorphic DNA Analysis for Genetic Characterization of Two Breeds of Dogs, German Shepherd and Japanese Spitz

2013 ◽  
Vol 13 (2) ◽  
pp. 73-78
Author(s):  
Jarina Joshsi ◽  
Lumanti Manandhar ◽  
Patima Shrestha ◽  
Rani Gupta ◽  
Rojlina Manadhar ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Rapd Markers ◽  
Dna Analysis ◽  
Genetic Characterization ◽  
Random Amplified Polymorphic Dna ◽  
Neighbor Joining ◽  
German Shepherd ◽  
Polymorphic Loci ◽  
Random Amplification

Random amplified polymorphic DNA (RAPD) markers were used to study genetic diversity in dog samples belonging to populations of German Shepherd and Japanese Spitz. A total of twelve samples were typed using eight RAPD primers. Out of eight primers, three primers gave result in six individuals of dogs. The phylogenetic tree constructed by the neighbor joining method based on Nei. Original measures revealed highest genetic identity found in German Shepherd as 0.9444 and highest genetic distance as 1.2809. The analysis predicts the number of polymorphic loci as 15 and the percentage of polymorphic loci as 83.3. Nepal Journal of Science and Technology Vol. 13, No. 2 (2012) 73-78 DOI: http://dx.doi.org/10.3126/njst.v13i2.7717

scholarly journals Isolation and Characterization of Two Klebsiella pneumoniae Phages Encoding Divergent Depolymerases

2020 ◽  
Vol 21 (9) ◽  
pp. 3160 ◽  
Author(s):  
Pilar Domingo-Calap ◽  
Beatriz Beamud ◽  
Lucas Mora-Quilis ◽  
Fernando González-Candelas ◽  
Rafael Sanjuán
Keyword(s):  
Klebsiella Pneumoniae ◽  
Multidrug Resistant ◽  
Health Concern ◽  
Resistant Bacteria ◽  
Isolation And Characterization ◽  
The Family ◽  
Important Challenge ◽  
Tail Spike ◽  
Reference Strains

The emergence of multidrug-resistant bacteria is a major global health concern. The search for new therapies has brought bacteriophages into the spotlight, and new phages are being described as possible therapeutic agents. Among the bacteria that are most extensively resistant to current antibiotics is Klebsiella pneumoniae, whose hypervariable extracellular capsule makes treatment particularly difficult. Here, we describe two new K. pneumoniae phages, πVLC5 and πVLC6, isolated from environmental samples. These phages belong to the genus Drulisvirus within the family Podoviridae. Both phages encode a similar tail spike protein with putative depolymerase activity, which is shared among other related phages and probably determines their ability to specifically infect K. pneumoniae capsular types K22 and K37. In addition, we found that phage πVLC6 also infects capsular type K13 and is capable of striping the capsules of K. pneumoniae KL2 and KL3, although the phage was not infectious in these two strains. Genome sequence analysis suggested that the extended tropism of phage πVLC6 is conferred by a second, divergent depolymerase. Phage πVLC5 encodes yet another putative depolymerase, but we found no activity of this phage against capsular types other than K22 and K37, after testing a panel of 77 reference strains. Overall, our results confirm that most phages productively infected one or few Klebsiella capsular types. This constitutes an important challenge for clinical applications.

Isolation and Characterization of Soil Myxobacteria from Nepal

2019 ◽  
Vol 24 (2) ◽  
pp. 7-16
Author(s):  
Nabin Rana ◽  
Saraswoti Khadka ◽  
Bishnu Prasad Marasini ◽  
Bishnu Joshi ◽  
Pramod Poudel ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Antimicrobial Properties ◽  
Antimicrobial Activities ◽  
Rrna Gene ◽  
Isolation And Characterization ◽  
The Family ◽  
Global Concern ◽  
Bacteria And Fungi ◽  
Antimicrobial Metabolites

 Realizing myxobacteria as a potential source of antimicrobial metabolites, we pursued research to isolate myxobacteria showing antimicrobial properties. We have successfully isolated three strains (NR-1, NR-2, NR-3) using the Escherichia coli baiting technique. These isolates showed typical myxobacterial growth characteristics. Phylogenetic analysis showed that all the strains (NR-1, NR-2, NR-3) belong to the family Archangiaceae, suborder Cystobacterineae, and order Myxococcales. Furthermore, 16S rRNA gene sequence similarity searched through BLAST revealed that strain NR-1 showed the closest similarity (91.8 %) to the type strain Vitiosangium cumulatum (NR-156939), NR-2 showed (98.8 %) to the type of Cystobacter badius (NR-043940), and NR-3 showed the closest similarity (83.5 %) to the type of strain Cystobacter fuscus (KP-306730). All isolates showed better growth in 0.5-1 % NaCl and pH around 7.0, whereas no growth was observed at pH 9.0 and below 5.0. All strains showed better growth at 32° C and hydrolyzed starch, whereas casein was efficiently hydrolyzed by NR-1 and NR-2. Besides, preliminary antimicrobial tests from crude extracts showed activities against Gram-positive, Gram-negative bacteria, and fungi. Our findings suggest that the arcane soil habitats of Nepal harbor myxobacteria with the capability to produce diverse antimicrobial activities that may be explored to overcome the rapidly rising global concern about antibiotic resistance.

Isolation and characterization of rad51 orthologs from Coprinus cinereus and Lycopersicon esculentum, and phylogenetic analysis of eukaryotic recA homologs

1997 ◽  
Vol 31 (2) ◽  
pp. 144-157 ◽  
Author(s):  
Natalie Yeager Stassen ◽  
J. M. Logsdon Jr. ◽  
G. J. Vora ◽  
Hildo H. Offenberg ◽  
Jeffrey D. Palmer ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Lycopersicon Esculentum ◽  
Coprinus Cinereus ◽  
Isolation And Characterization

Cloning and expression of DiT33 from Dirofilaria immitis: a specific and early marker of heartworm infection

Parasitology ◽  
1996 ◽  
Vol 112 (3) ◽  
pp. 331-338 ◽  
Author(s):  
X. Q. Hong ◽  
J. Santiago Mejia ◽  
S. Kumar ◽  
F. B. Perler ◽  
C. K. S. Carlow
Keyword(s):  
Dirofilaria Immitis ◽  
Cloning And Expression ◽  
E Coli ◽  
Spliced Leader ◽  
Isolation And Characterization ◽  
The Family ◽  
Heartworm Infection ◽  
High Level ◽  
Control And Management

SUMMARYDirofilaria immitis is an important filarial parasite of dogs and cats, and a useful model for human filariasis. Current diagnostic tests for heartworm infection in animals rely on the presence of fecund female worms (usually found 6·5 months post-infection or later) and therefore fail to detect pre-patent infections. Putative pepsin inhibitors from 2 filarial parasites of humans namely Onchocerca volvulus (Ov33, Oc3.6, OvDSB) and Brugia malayi (Bm33), have been shown to be useful in diagnosis of onchocerciasis and lymphatic filariasis, respectively. Previous studies have suggested that a homologue exists in D. immitis (DiT33), which may have potential in diagnosis of heartworm infection. In this study, the isolation and characterization of a cDNA clone encoding DiT33 is described.‡ This cDNA contains 12 bases of the nematode-specific 22 nucleotide spliced leader sequence and encodes a 26·4 kDa-protein with a high level of similarity (87–89%) to other filarial members of the family. DJT33 was over-expressed in E. coli as a fusion with the maltose-binding protein and serological analysis was performed using a panel of clinically defined dog sera. The findings of this study indicate that DiT33 is a promising antigen for the early detection of D. immitis and may be a valuable tool in the control and management of heartworm infection.

scholarly journals Isolation and characterization of hantavirus carried by Apodemus peninsulae in Jilin, China

2007 ◽  
Vol 88 (4) ◽  
pp. 1295-1301 ◽  
Author(s):  
Yong-Zhen Zhang ◽  
Yang Zou ◽  
Lai-Shun Yao ◽  
Guang-Wei Hu ◽  
Zhan-Shen Du ◽  
...  
Keyword(s):  
Cell Culture ◽  
Phylogenetic Analysis ◽  
Eastern China ◽  
Immunofluorescence Assay ◽  
Haemorrhagic Fever ◽  
Apodemus Peninsulae ◽  
Isolation And Characterization ◽  
North Eastern ◽  
Field Mice

To provide a better understanding of hantavirus epidemiology in China, Korean field mice (Apodemus peninsulae) and striped field mice (Apodemus agrarius) were captured in Jilin province, China, where haemorrhagic fever with renal syndrome (HFRS) is endemic. Hantavirus antigens were detected in eight of the 130 A. peninsulae individuals and in four of the 193 A. agrarius individuals by using an immunofluorescence assay. Partial S and M segments were amplified from all of the antigen-positive samples. Furthermore, two hantaviruses (CJAp89 and CJAp93) were isolated successfully in cell culture and the entire S and M segments were amplified from one of them (CJAp93). Phylogenetic analysis of these sequences (partial or complete) showed that hantaviruses carried by A. peninsulae and A. agrarius form two distinct lineages, although viruses carried by A. peninsulae are similar to those isolated previously from A. agrarius in China and from HFRS patients in Russia. However, the viruses detected in A. peninsulae in China are genetically different from those detected in A. peninsulae in other countries. These data suggest that A. peninsulae is also a natural host for HTNV in north-eastern China.

scholarly journals Isolation and Characterization of Influenza C Viruses in the Philippines and Japan

2014 ◽  
Vol 53 (3) ◽  
pp. 847-858 ◽  
Author(s):  
Takashi Odagiri ◽  
Yoko Matsuzaki ◽  
Michiko Okamoto ◽  
Akira Suzuki ◽  
Mariko Saito ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Mdck Cells ◽  
Severe Pneumonia ◽  
Respiratory Illness ◽  
The Philippines ◽  
Antigenic Analysis ◽  
Isolation And Characterization ◽  
Gene Sequence Analysis ◽  
Influenza C Virus

From November 2009 to December 2013 in the Philippines, 15 influenza C viruses were isolated, using MDCK cells, from specimens obtained from children with severe pneumonia and influenza-like illness (ILI). This is the first report of influenza C virus isolation in the Philippines. In addition, from January 2008 to December 2013, 7 influenza C viruses were isolated from specimens that were obtained from children with acute respiratory illness (ARI) in Sendai city, Japan. Antigenic analysis with monoclonal antibodies to the hemagglutinin-esterase (HE) glycoprotein showed that 19 strains (12 from the Philippines and 7 from Japan) were similar to the influenza C virus reference strain C/Sao Paulo/378/82 (SP82). Phylogenetic analysis of the HE gene showed that the strains from the Philippines and Japan formed distinct clusters within an SP82-related lineage. The clusters that included the Philippine and Japanese strains were shown to have diverged from a common ancestor around 1993. In addition, phylogenetic analysis of the internal genes showed that all strains isolated in the Philippines and Japan had emerged through reassortment events. The composition of the internal genes of the Philippine strains was different from that of the Japanese strains, although all strains were classified into an SP82-related lineage by HE gene sequence analysis. These observations suggest that the influenza C viruses analyzed here had emerged through different reassortment events; however, the time and place at which the reassortment events occurred were not determined.

Isolation and characterization of microsatellites inNeogobius kessleri(Perciformes, Gobiidae) and cross-species amplification within the family Gobiidae

2007 ◽  
Vol 7 (4) ◽  
pp. 701-704 ◽  
Author(s):  
MARTINA VYSKOČILOVÁ ◽  
MARKÉTA ONDRAČKOVÁ ◽  
ANDREA ŠIMKOVÁ ◽  
JEAN-FRANÇOIS MARTIN
Keyword(s):  
Isolation And Characterization ◽  
The Family

Isolation and characterization of bacteriophages specific to hydrogen-sulfide-producing bacteria

2013 ◽  
Vol 59 (1) ◽  
pp. 39-45 ◽  
Author(s):  
Chao Gong ◽  
Spencer Heringa ◽  
Randhir Singh ◽  
Jinkyung Kim ◽  
Xiuping Jiang
Keyword(s):  
Hydrogen Sulfide ◽  
Lag Phase ◽  
Restriction Enzyme Digestion ◽  
Isolation And Characterization ◽  
The Family ◽  
Processing Plants ◽  
Specific Bacteriophage ◽  
Myoviridae Family ◽  
Restriction Enzyme Digestion Analysis

The objectives of this study were to isolate and characterize bacteriophages specific to hydrogen-sulfide-producing bacteria (SPB) from raw animal materials, and to develop a SPB-specific bacteriophage cocktail for rendering application. Meat, chicken offal, and feather samples collected from local supermarkets and rendering processing plants were used to isolate SPB (n = 142). Bacteriophages (n = 52) specific to SPB were isolated and purified from the above samples using 18 of those isolated SPB strains as hosts. The host ranges of bacteriophages against 5 selected SPB strains (Escherichia coli, Citrobacter freundii, and Hafnia alvei) were determined. Electron microscopy observation of 9 phages selected for the phage cocktail revealed that 6 phages belonged to the family of Siphoviridae and 3 belonged to the Myoviridae family. Restriction enzyme digestion analysis with endonuclease DraI detected 6 distinguished patterns among the 9 phages. Phage treatment prevented the growth of SPB for up to 10 h with multiplicity of infection ratios of 1, 10, 100, and 1000 in tryptic soy broth at 30 °C, and extended the lag phase of SPB growth for 2 h at 22 °C with multiplicities of infection of 10, 100, and 1000. These results suggest that the selected bacteriophage cocktail has a high potential for phage application to control SPB in raw animal materials destined for the rendering process.

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