scholarly journals Induction of Multiple Prophages from a Marine Bacterium: a Genomic Approach

2006 ◽  
Vol 72 (7) ◽  
pp. 4995-5001 ◽  
Author(s):  
Feng Chen ◽  
Kui Wang ◽  
Jeneen Stewart ◽  
Robert Belas
Keyword(s):  
Mitomycin C ◽  
Marine Bacterium ◽  
Pcr Amplification ◽  
Pulsed Field Gel Electrophoresis ◽  
Bacterial Genomes ◽  
Dominant Type ◽  
Transmission Electron ◽  
Viral Particles ◽  
Genomic Approach ◽  
Primer Sets

ABSTRACT Approximately 70% of sequenced bacterial genomes contain prophage-like structures, yet little effort has been made to use this information to determine the functions of these elements. The recent genomic sequencing of the marine bacterium Silicibacter sp. strain TM1040 revealed five prophage-like elements in its genome. The genomes of these prophages (named prophages 1 to 5) are approximately 74, 30, 39, 36, and 15 kb long, respectively. To understand the function of these prophages, cultures of TM1040 were treated with mitomycin C to induce the production of viral particles. A significant increase in viral counts and a decrease in bacterial counts when treated with mitomycin C suggested that prophages were induced from TM1040. Transmission electron microscopy revealed one dominant type of siphovirus, while pulsed-field gel electrophoresis demonstrated two major DNA bands, equivalent to 35 and 75 kb, in the lysate. PCR amplification with primer sets specific to each prophage detected the presence of prophages 1, 3, and 4 in the viral lysate, suggesting that these prophages are inducible, but not necessarily to the same level, while prophages 2 and 5 are likely defective or non-mitomycin C-inducible phages. The combination of traditional phage assays and modern microbial genomics provides a quick and efficient way to investigate the functions and inducibility of prophages, particularly for a host harboring multiple prophages with similar sizes and morphological features.

EXPLORING THE CHRONIC MORTALITY AFFECTING ABALONES IN TAIWAN: DIFFERENTIATION OF ABALONE HERPESVIRUS-ASSOCIATED ACUTE INFECTION FROM CHRONIC MORTALITY BY PCR AND IN SITU HYBRIDIZATION AND HISTOPATHOLOGY

2016 ◽  
Vol 42 (01) ◽  
pp. 1-9 ◽  
Author(s):  
I-Wen Chen ◽  
Pen-Heng Chang ◽  
Min-Shou Chen ◽  
Tristan Renault ◽  
Meei-Mei Chen ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Acute Infection ◽  
Extended Period ◽  
Haliotis Diversicolor Supertexta ◽  
Transmission Electron ◽  
Viral Particles ◽  
Primer Sets ◽  
Diversicolor Supertexta ◽  
Pcr In Situ

Abalone herpesvirus (AbHV) infection of cultured abalones Haliotis diversicolor supertexta induced acute high mortality in 2003. Years later, sporadic mortality was noted for an extended period of months, resulting in high cumulative mortality. Moribund abalones were analyzed using PCR, in situ hybridization, and histopathology, because thus far no viral particles have been observed by transmission electron microscopy. PCR using 20 primer sets, specifically designed from sequences of acute AbHV infection, failed to amplify any products from abalones suffering from chronic mortality. Subsequently, a 1406-bp sequence was amplified from chronic moribund abalones, and this sequence showed a 92% (553 bp/602 bp) homology with the gene of an AbHV Taiwan isolate (NCBI serial no. KF537536.1), suggestive of an AbHV pathotype. Histopathology of AbHV pathotype infection showed hemocyte infiltration in the lamina propia of the digestive tract, and hemocytes of various stages were evident, as well as the loss of seminal tubules in the gonad. In situ hybridization revealed that in AbHV infection, positive signals were restricted to the neural ganglia, while in AbHV pathotype infection, positive signals were observed only in the hemocytes. It appeared that the tropism of AbHV shifted from mainly neurotropic in AbHV infection to mainly hemocytotropic in abalone suffering from chronic mortality. Abalone shriveling syndrome-associated virus co-infection was detected in some of AbHV pathotype infection events. Further studies are needed to better understand the pathogenesis of AbHV pathotype affecting H. diversicolor in Taiwan.

Marinomonas mediterranea is a lysogenic bacterium that synthesizes R-bodies

Microbiology ◽  
2003 ◽  
Vol 149 (9) ◽  
pp. 2679-2686 ◽  
Author(s):  
Diana Hernández-Romero ◽  
Patricia Lucas-Elío ◽  
Daniel López-Serrano ◽  
Francisco Solano ◽  
Antonio Sanchez-Amat
Keyword(s):  
Mitomycin C ◽  
Marine Bacterium ◽  
Sea Water ◽  
Free Living ◽  
Gene Encoding ◽  
Transmission Electron ◽  
Extrachromosomal Elements ◽  
Lysogenic Bacterium ◽  
Resistant Strains ◽  
Micro Organisms

The melanogenic marine bacterium Marinomonas mediterranea synthesizes R-bodies as revealed by transmission electron microscopy. These structures were previously described in some obligate symbionts of paramecia and some free-living bacteria, none of which was isolated from sea water. In other micro-organisms, the synthesis of R-bodies has been related to extrachromosomal elements. Accordingly, M. mediterranea induction by mitomycin C or UV radiation resulted in the production of defective phages resembling bacteriocins, indicating that it is a lysogenic bacterium. Two mitomycin-C-resistant strains defective in prophage replication have been isolated. These mutants, and the previously obtained strains ngC1, T102 and T103, the latter mutated in the ppoS gene encoding a sensor histidine kinase, are affected not only in phage replication but also in polyphenol oxidase activities and melanin synthesis, suggesting a relationship between the control of all these processes.

A combined pseudoreplica-immunocytochemical technique for research and diagnostic virology

1990 ◽  
Vol 48 (3) ◽  
pp. 900-901
Author(s):  
Blayne Fritz ◽  
Stanley J. Naides ◽  
Kenneth Moore
Keyword(s):  
Phosphotungstic Acid ◽  
Immunogold Labelling ◽  
Uranyl Acetate ◽  
Distilled Water ◽  
Clinical Specimens ◽  
Tissue Sections ◽  
Specimen Retrieval ◽  
Transmission Electron ◽  
Viral Particles ◽  
Diagnostic Virology

The pseudoreplica method of staining viral particles for visualization by transmission electron microscopy is a very popular technique. The ability to concentrate clinical specimens while semi-embedding viral particles makes it especially well suited for morphologic and diagnostic virology. Immunolabelling viral particles with colloidal gold is a technique frequently employed by both research and diagnostic virologists. We have characterized a procedure which provides the advantage of both by modifying and combining pseudoreplica staining and immunogold labelling.Modification of specimen retrieval and delay of staining allows us to utilize pseudoreplica processed specimens within our standard immunogold labelling protocol. In brief, we absorbed samples onto 2% agarose, added.25% Formvar and wicked dry. We then floated the Formvar-virus film onto double distilled water, added grids and retrieved with parafilm. The Formvar-virus specimens were then treated as thin tissue sections within our standard two stage immunolabelling protocol. Following completion of immunogold labelling; each grid was negatively stained with phosphotungstic acid or uranyl acetate contrast stains.

Are there Viruses in Industrial Bioleaching Econiches?

2017 ◽  
Vol 262 ◽  
pp. 521-525 ◽  
Author(s):  
Paulo C. Covarrubias ◽  
Rodrigo Muñoz ◽  
Roberto A. Bobadilla-Fazzini ◽  
Patricio Martinez ◽  
Raquel Quatrini
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Microbial Ecology ◽  
Mineral Processing ◽  
Microbial Consortia ◽  
Basic Question ◽  
Host Interactions ◽  
Transmission Electron ◽  
Viral Particles ◽  
Stability Issues

Detailed descriptions of the consortia present in commercial mineral processing operations have emerged in recent years, improving our understanding of the biology and the ecology of bioleaching. In spite of this progress, one of the aspects of biomining microbial ecology that remains un-tackled is that of virus-host interactions. The effects of viruses on the dynamics of the bioleaching microbial consortia and their impact in metal recovery is presently unknown. To begin addressing this issue we asked a basic question: ¿Are there viruses in industrial bioleaching econiches? In this work, we answer that question experimentally, assessing the number and types of viral particles recovered in the leachates from different industrial settings, using epifluorescence and transmission electron microscopy. Findings emerging from this work point to an almost null presence of viral particles in the leachates from mineral processing operations, possibly due to structural stability issues of the particles in the extreme acidic and highly oxidant conditions favoured by their potential microbial hosts. In turn, DNA-loaded viral-size vesicles of presently unknown function are frequent and abundant in all samples analysed.

scholarly journals Structure and molecular analysis of RGR1, a gene required for glucose repression of Saccharomyces cerevisiae.

1990 ◽  
Vol 10 (8) ◽  
pp. 4130-4138 ◽  
Author(s):  
A Sakai ◽  
Y Shimizu ◽  
S Kondou ◽  
T Chibazakura ◽  
F Hishinuma
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gel Electrophoresis ◽  
Daughter Cell ◽  
Glucose Repression ◽  
Pulsed Field Gel Electrophoresis ◽  
Null Mutation ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Transmission Electron Microscopic ◽  
Carboxy Terminal

An RGR1 gene product is required to repress expression of glucose-regulated genes in Saccharomyces cerevisiae. The abnormal morphology of rgr1 cells was studied. Scanning and transmission electron microscopic observations revealed that the cell wall of the daughter cell remained attached to that of mother cell. We cloned the RGR1 gene by complementation and showed that the cloned DNA was tightly linked to the chromosomal RGR1 locus. The cloned RGR1 gene suppressed all of the phenotypes caused by the mutation and encoded a 3.6-kilobase poly(A)+ RNA. The RGR1 gene is located on chromosome XII, as determined by pulsed-field gel electrophoresis, and we mapped rgr1 between gal2 and pep3 by genetic analysis. rgr1 was shown to be a new locus. We also determined the nucleotide sequence of RGR1, which was predicted to encode a 123-kilodalton protein. The null mutation resulted in lethality, indicating that the RGR1 gene is essential for growth. On the other hand, a carboxy-terminal deletion of the gene caused phenotypes similar to but more severe than those caused by the original mutation. The amount of reserve carbohydrates was reduced in rgr1 cells. Possible functions of the RGR1 product are discussed.

scholarly journals Mitomycin C Inhibits Esophageal Fibrosis by Regulating Cell Apoptosis and Autophagy via lncRNA-ATB and miR-200b

2021 ◽  
Vol 8 ◽  
Author(s):  
Yin Zhang ◽  
Qinge Wang ◽  
Yuping Xu ◽  
Jing Sun ◽  
Yanbo Ding ◽  
...  
Keyword(s):  
Electron Microscope ◽  
Mitomycin C ◽  
Cell Apoptosis ◽  
Optimal Dose ◽  
The Other ◽  
Fibroblast Cells ◽  
Transmission Electron ◽  
Esophageal Strictures ◽  
Related Proteins

Benign esophageal strictures (BESs) frequently results from esophageal fibrosis. The transformation of fibroblasts into fibrocyte is an important cause of fibrosis. The treatment of fibrosis is challenging. Some previous studies have indicated the antifibrotic effect of mitomycin C (MMC). However, the mechanism of action of MMC and its optimal dose for treatment remains unclear. In the present study, the role of MMC in fighting fibrosis and its mechanism was investigated. Human esophageal fibroblast cells (HEFs)were treated without or with MMC, at 2, 5, 10 μg/ml, combining with mimic lncRNA-ATB, miR-200b inhibitor, rapamycin (RAPA), and 3-Methyladenine (3-MA). The cell viability, and cell apoptosis were evaluated. In addition, expression of apoptosis related proteins (caspase8 and caspase3), autophagy related proteins (LC3II and ATG5) and fibrosis related proteins (α-SMA collagen-1 and TGF-β) were also evaluated. Furthermore, autophagosome was observed by transmission electron microscope. Results showed that the expression of lncRNA-ATB was down-regulated and miR-200b was up-regulated after treated with MMC. And MMC induced cell apoptosis and inhibited cell autophagy. On the other hand, RAPA, mimic lncRNA-ATB and miR-200b inhibitor reduced fibrogenic effect of MMC on HEFs. Collectively, this study suggests that MMC inhibited esophageal fibrosis by regulating cell apoptosis and autophagy via downregulating lncRNA-ATB and upregulating miR-200b.

scholarly journals Cellular tropism of SARS-CoV-2 in the respiratory tract of Syrian hamsters and B6.Cg-Tg(K18-ACE2)2Prlmn/J transgenic mice

2021 ◽  
pp. 030098582110430
Author(s):  
Hui-Ling Yen ◽  
Sophie Valkenburg ◽  
Sin Fun Sia ◽  
Ka Tim Choy ◽  
J. S. Malik Peiris ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Lung Parenchyma ◽  
Productive Infection ◽  
Type I ◽  
Syrian Hamsters ◽  
Transmission Electron ◽  
Type Ii Pneumocytes ◽  
Viral Particles ◽  
Cellular Tropism ◽  
Conducting Airways

Several animal models have been developed to study the pathophysiology of SARS-CoV-2 infection and to evaluate vaccines and therapeutic agents for this emerging disease. Similar to infection with SARS-CoV-1, infection of Syrian hamsters with SARS-CoV-2 results in moderate respiratory disease involving the airways and lung parenchyma but does not lead to increased mortality. Using a combination of immunohistochemistry and transmission electron microscopy, we showed that the epithelium of the conducting airways of hamsters was the primary target for viral infection within the first 5 days of infection, with little evidence of productive infection of pneumocytes. At 6 days postinfection, antigen was cleared but parenchymal damage persisted, and the major pathological changes resolved by day 14. These findings are similar to those previously reported for hamsters with SARS-CoV-1 infection. In contrast, infection of K18-hACE2 transgenic mice resulted in pneumocyte damage, with viral particles and replication complexes in both type I and type II pneumocytes together with the presence of convoluted or cubic membranes; however, there was no evidence of virus replication in the conducting airways. The Syrian hamster is a useful model for the study of SARS-CoV-2 transmission and vaccination strategies, whereas infection of the K18-hCE2 transgenic mouse results in lethal disease with fatal neuroinvasion but with sparing of conducting airways.

scholarly journals Canine bocavirus-2 infection and its possible association with encephalopathy in domestic dogs

PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0255425
Author(s):  
Chutchai Piewbang ◽  
Sabrina Wahyu Wardhani ◽  
Wichan Dankaona ◽  
Sitthichok Lacharoje ◽  
Poowadon Chai-in ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Control Group ◽  
Normal Brain ◽  
Evolutionary Analysis ◽  
Histological Changes ◽  
Brain Infection ◽  
Transmission Electron ◽  
Viral Particles ◽  
Cellular Tropism

Canine bocaviruses (CBoVs) have been recognized as pathogens associated with intestinal diseases. Hematogenous spreading caused by CBoV has been documented and may potentiate the virus entry across the blood-brain barrier to initiate a brain infection. This study focused attention on CBoV detection in cases of encepahlopathy and attempted to determine its viral localization. A total of 107 dog brains that histologically exhibited encephalopathy (ED) were investigated for the presence of CBoVs using polymerase chain reaction (PCR). Thirty-three histologically normal brain samples from dogs were used as a control group (CD). CBoV-2 was detected in 15 ED dogs (14.02%) but not in CD dogs (p = 0.02), while no CBoV-1 and -3 were detected. Among the CBoV-2 positive dogs, brain histological changes were characterized by nonsuppurative encephalitis, with inclusion body-like materials in some brains. In situ hybridization (ISH) and transmission electron microscopy (TEM) confirmed the presence of CBoV-2 viral particles in glial cells, supporting neurotropism of this virus. ISH signals were also detected in the intestines, lymphoid organs, and the heart, suggesting both enteral and parenteral infections of this virus. Whole genome characterization and evolutionary analysis revealed genetic diversity of CBoV-2 sequences and it was varying among the different countries where the virus was detected. This study points to a possible association of CBoV-2 with encephalopathy in dogs. It also highlights the genetic diversity and cellular tropism of this virus.

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