Detection and isolation of Duck Plague virus from field outbreaks in Assam, India

2018 ◽  
Author(s):  
Samsun Neher ◽  
Nagendra Nath Barman ◽  
Durlav Prasad Bora ◽  
Dipak Deka ◽  
Shantanu Tamuly ◽  
...  
Keyword(s):  
Cell Culture ◽  
Dna Polymerase ◽  
Primary Cell Culture ◽  
Present Report ◽  
Sandwich Elisa ◽  
Post Mortem ◽  
Polymerase Gene ◽  
Dna Virus ◽  
Internal Organs ◽  
Duck Plague Virus

Duck plague is an acute, contagious and lethal disease of ducks caused by an enveloped DNA virus, belonging to Anatid Herpes Virus 1 under genus Mardivirus, subfamily Alphaherpesvirinae, family Herpesviridae. The disease is characterized by sudden death, high mortality, hemorrhages and necrosis in the internal organs. Duck farming plays an important role in the livelihood of the farmers of Assam. Duck Plague being the major killer disease among duck population of Assam, posing a threat to the growth of duck farming in the state. The present report investigates various outbreaks of Duck plague in Assam, detection and isolation of the virus. A total of 29 outbreaks suspected for Duck Plague were attended in different districts of Assam and 380 numbers of samples comprising of both clinical (n=107) as well as post mortem (n=273) were collected. Presence of DPV in samples was detected by Sandwich-ELISA (S-ELISA) and amplification of UL 44.5 as well as DNA polymerase gene of DPV by PCR. Out of 380 samples collected from 29 outbreaks, 213 (56.05%) and 299 (78.68%) samples were found positive for presence of DPV by S-ELISA and PCR, respectively. Pooled positive samples from various outbreaks were processed for isolation of duck plague virus in DEF primary cell culture. DPV field strain produced CPE comprising of rounding of cells, formation of syncytia, marked cytoplasmic granulations and intracellular vacuoles.

scholarly journals Molecular Characterization of Duck Plague Virus for Determination of TCID50

2021 ◽  
Vol 8 (1) ◽  
pp. 125-133
Author(s):  
Md Salim Jahan ◽  
Md Mizanur Rahman ◽  
Rony Ahmed ◽  
Ajran Kabir ◽  
ShamsulKaunain Oli ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Molecular Characterization ◽  
Dna Polymerase ◽  
Molecular Detection ◽  
Chorioallantoic Membrane ◽  
Post Mortem ◽  
Duck Embryo Fibroblast ◽  
Post Mortem Examination ◽  
Duck Embryo ◽  
Duck Plague Virus

Duck plague is an enveloped DNA virus that belongs to the Anatid Herpes Virus; the Herpesviridae family is an acute and highly infectious duck, geese, and swan disease that causes tremendous economic losses of duck rearing in Bangladesh and other duck rearing countries. Therefore, we decided to isolate duck plague virus from recent fields’ outbreaks area and performed molecular detection and phylogenetic analysis to find out the similarities between our findings and other isolates around the world. Visceral organs of 13 suspected ducks from recent outbreaks area were collected by post-mortem examination for inoculum preparation. Several passages were performed to harvest into 9-11 old embryonated eggs Chorioallantoic membrane (CAM) route and duck embryo fibroblast (DEF) primary cell culture. DNA polymerase (446bp) and DNA polymerase (UL, 602bp) genes were used for molecular detection by Polymerase chain reaction (PCR). Pathogenicity was done with duckling and TCID50 on DEF. Molecular characterization was performed from extracted DNA of duckling and 2 Positive PCR products were partially sequenced for phylogenetic analysis of their origin and nucleotide variations. Sequenced data was analyzed to reveal genetic relationships among constructed phylogenetic tree for understanding potential transmission with origin of virus and data was then submitted to gene bank and got accession number for DPV-BR1-MN937272 and DPV-BR-2-MN937273. Among 13 samples, 4(30.77%) were found positive by PCR using DNA polymerase at 446 bp and UL at 602 bp gene. Chorioallantoic membrane (CAM) was observed hemorrhagic after 72 days and duck embryo fibroblast (DEF) become round as showed cytopathic characteristics after 48h of infection .Duckling showed that isolated virus was highly pathogenic as characteristics signs of post-mortem examination. Therefore, this has found that recent isolates have similarity with Bangladesh, India and China isolates. Moreover, TCID50 has confirmed the isolates have accepted titer to be a vaccine strain. Res. Agric., Livest. Fish.8(1): 125-133, April 2021

The role of ER stress for the pathogenesis of SCA3 in primary cell culture experiment

2007 ◽  
Vol 34 (S 2) ◽  
Author(s):  
C Funke ◽  
J Hübener ◽  
H Wolburg ◽  
T Schmidt ◽  
H Toresson ◽  
...  
Keyword(s):  
Cell Culture ◽  
Er Stress ◽  
Primary Cell Culture ◽  
Primary Cell ◽  
Culture Experiment ◽  
Cell Culture Experiment

scholarly journals Genetic evidence for two protein domains and a potential new activity in bacteriophage T4 DNA polymerase.

Genetics ◽  
1990 ◽  
Vol 124 (2) ◽  
pp. 213-220 ◽  
Author(s):  
L J Reha-Krantz
Keyword(s):  
Dna Polymerase ◽  
Bacteriophage T4 ◽  
Rnase H ◽  
Ribonuclease H ◽  
Temperature Sensitive ◽  
Polymerase Gene ◽  
Dna Polymerase I ◽  
E Coli ◽  
Intragenic Complementation ◽  
Polymerase I

Abstract Intragenic complementation was detected within the bacteriophage T4 DNA polymerase gene. Complementation was observed between specific amino (N)-terminal, temperature-sensitive (ts) mutator mutants and more carboxy (C)-terminal mutants lacking DNA polymerase polymerizing functions. Protein sequences surrounding N-terminal mutation sites are similar to sequences found in Escherichia coli ribonuclease H (RNase H) and in the 5'----3' exonuclease domain of E. coli DNA polymerase I. These observations suggest that T4 DNA polymerase, like E. coli DNA polymerase I, contains a discrete N-terminal domain.

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