scholarly journals Integrated Use of Molecular Techniques to Detect and Genetically Characterise DNA Viruses in Italian Wolves (Canis lupus italicus)

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2198
Author(s):  
Andrea Balboni ◽  
Lorenza Urbani ◽  
Mauro Delogu ◽  
Carmela Musto ◽  
Maria Cristina Fontana ◽  
...  
Keyword(s):  
Canis Lupus ◽  
Viral Genome ◽  
Molecular Techniques ◽  
Internal Organs ◽  
Vp2 Gene ◽  
Dna Viruses ◽  
Viral Genes ◽  
Pcr Assays ◽  
Free Ranging ◽  
Complete Viral Genome

In this study, internal organs (tongue, intestine, and spleen) of 23 free-ranging Italian wolves (Canis lupus italicus) found dead between 2017 and 2019 were tested for Carnivore protoparvovirus 1, Canine adenovirus (CAdV), and Canine circovirus (CanineCV) using real-time PCR assays. Genetic characterisation of the identified viruses was carried out by amplification, sequencing, and analysis of the complete viral genome or informative viral genes. All the wolves tested positive for at least one of the DNA viruses screened, and 11/23 were coinfected. Carnivore protoparvoviruses were the most frequently detected viruses (21/23), followed by CanineCV (11/23) and CAdV (4/23). From the analysis of the partial VP2 gene of 13 carnivore protoparvoviruses, 12 were canine parvovirus type 2b, closely related to the strains detected in dogs and wild carnivores from Italy, and one was a feline panleukopenia-like virus. Of the four CAdV identified, two were CAdV-1 and two were CAdV-2. The complete genome of seven CanineCVs was sequenced and related to the CanineCV identified in dogs, wolves, and foxes worldwide. Close correlations emerged between the viruses identified in wolves and those circulating in domestic dogs. Further studies are needed to investigate if these pathogens may be potentially cross-transmitted between the two species.

scholarly journals Latest Advances of Virology Research Using CRISPR/Cas9-Based Gene-Editing Technology and Its Application to Vaccine Development

Viruses ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 779
Author(s):  
Man Teng ◽  
Yongxiu Yao ◽  
Venugopal Nair ◽  
Jun Luo
Keyword(s):  
Biological Sciences ◽  
Gene Therapy ◽  
Genetic Engineering ◽  
Gene Function ◽  
Viral Genome ◽  
Gene Editing ◽  
Vaccine Development ◽  
Future Prospects ◽  
Dna Viruses ◽  
Viral Genomes

In recent years, the CRISPR/Cas9-based gene-editing techniques have been well developed and applied widely in several aspects of research in the biological sciences, in many species, including humans, animals, plants, and even in viruses. Modification of the viral genome is crucial for revealing gene function, virus pathogenesis, gene therapy, genetic engineering, and vaccine development. Herein, we have provided a brief review of the different technologies for the modification of the viral genomes. Particularly, we have focused on the recently developed CRISPR/Cas9-based gene-editing system, detailing its origin, functional principles, and touching on its latest achievements in virology research and applications in vaccine development, especially in large DNA viruses of humans and animals. Future prospects of CRISPR/Cas9-based gene-editing technology in virology research, including the potential shortcomings, are also discussed.

scholarly journals Evaluation of Sandwich Enzyme-Linked Immunosorbent Assay and Reverse Transcription Polymerase Chain Reaction for the Diagnosis of Foot-and-Mouth Disease

Intervirology ◽  
2021 ◽  
pp. 1-6
Author(s):  
Salman Khan ◽  
Syed Asad Ali Shah ◽  
Syed Muhammad Jamal
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Genome ◽  
Foot And Mouth Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Kappa Value ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Level Of Agreement

<b><i>Background:</i></b> Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. <b><i>Methods:</i></b> A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. <b><i>Results:</i></b> S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of the samples using pan-FMDV primers set, 1F/1R. Thirty-six samples were found positive and 7 negative by both the tests. The level of agreement between the tests was assessed by calculating the Kappa value, which was found to be fair (Kappa value = 0.303 and 95% CI = 0.089; 0.517) and significant (<i>p</i> = 0.009). However, 2 samples, which were found positive on S-ELISA tested negative on RT-PCR. This may be attributed to the presence of nucleotide mismatch(es) in the primer-binding sites that may have resulted in failure of amplification of the viral genome. The serotype-specific RT-PCR assays not only confirmed serotyping results of S-ELISA but were also able to establish serotype in 9 S-ELISA-negative but pan-FMDV RT-PCR-positive samples. <b><i>Conclusions:</i></b> The RT-PCR assay contributes significantly to establishing a quick, sensitive, and definitive diagnosis of FMD in resource-constrained countries. Samples giving negative results in S-ELISA should be tested in RT-PCR for the disease detection and virus typing.

scholarly journals Pathology and One Health implications of fatal Leptospira interrogans infection in an urbanized, free-ranging, black-tufted marmoset ( Callithrix penicilatta ) in Brazil

2021 ◽  
Author(s):  
Tais M. Wilson ◽  
Jana M. Ritter ◽  
Roosecelis B. Martines ◽  
Alexandra A. B. Gonçalves ◽  
Pamela Fair ◽  
...  
Keyword(s):  
One Health ◽  
Pulmonary Hemorrhage ◽  
Health Concern ◽  
Leptospira Interrogans ◽  
Public Health Concern ◽  
Wide Range ◽  
Health Related ◽  
Pcr Assays ◽  
Free Ranging ◽  
First Time

Leptospirosis is a zoonotic neglected disease of worldwide public health concern. Leptospira species can infect a wide range of wild and domestic mammals and can lead to a spectrum of disease, including severe and fatal forms. Herein, we report for the first time a fatal Leptospira interrogans infection in a free-ranging nonhuman primate (NHP), a black-tufted marmoset. Icterus, pulmonary hemorrhage, interstitial nephritis and hepatocellular dissociation were the main findings raising the suspicion of leptospirosis. Diagnostic confirmation was based on specific immunohistochemical and PCR assays for Leptospira species. Immunolocalization of leptospiral antigens and identification of pathogenic species ( L. interrogans species) were important for better understanding the pathogenesis of disease. One Health related implications of free-ranging NHPs in anthropized areas and transmission dynamics of human and animal leptospirosis are discussed.

Abstract 20433: Viral Myocarditis: Viral Genome Detection Yield on Endomyocardial Biopsy is Influenced by Methodological Factors

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Elisa Carturan ◽  
Stefania Rizzo ◽  
Gaetano Thiene ◽  
Cristina Basso
Keyword(s):  
Endomyocardial Biopsy ◽  
Viral Genome ◽  
Viral Myocarditis ◽  
Molecular Techniques ◽  
Time Interval ◽  
Tissue Fixation ◽  
Viral Genomes ◽  
Consensus Document ◽  
Significant Difference

Introduction: Myocarditis is diagnosed on endomyocardial biopsy (EMB) by histological, immune-histochemical and molecular techniques to detect viral genomes. The role of EMB for the diagnosis and its contribution to patient management has been addressed by a Consensus document of American and European Cardiovascular Pathology Societies. Hypothesis: Different methods of tissue fixation and the number of myocardial samples could impact on viral genome yield on EMB. Methods: Consecutive EMBs referred at our Institution for histology/ immunohistochemistry evaluation in the time interval 1996-2012 were enrolled. Molecular investigation by RT-PCR and PCR technique was performed in all EMBs with inflammatory cardiomyopathy diagnosis at histology and immunohistochemistry. Results: A total of 467 EMBs have been diagnosed as myocarditis: 79 in pediatric (1 month to 18 years; 47M/ 32F) and 388 adult (19 to 75 years; 256M/132 F) patients (pts.). Viral etiology was identified in 28 (36%) pediatric and 101 (26%) adult pts, and the most prevalent type of viruses were enterovirus (8/28, 36%; 26/101, 24%). In a more recent subgroup of 137 EMBs (virus positive 30/137, 22%) seasonality, type of fixation and number of EMB samples were assessed. The seasonal distribution of myocarditis was higher in winter than in other seasons (33% vs. autumn 28%, spring 20% and summer 19%) without any significant difference in terms of virus positive EMBs. The number of EMB samples per pt. was ≤3 (either formalin or RNAlater) in 94/137 (69%) and >3 in 43/137 (31%), with a lower prevalence of virus positive in the former (17/94, 18% vs. 13/43, 30%). Eighty-one EMB samples were frozen while 56 were paraffin embedded, with a higher prevalence of viral genome in the former (26/81, 32% vs. 4/56, 7%; p=0.001). Conclusions: The diagnosis of myocarditis on EMB samples requires standardized protocols including molecular techniques. Viral genomes are identifiable in more than one third of pediatric and one fourth of adult cases. Methodological factors like the type of tissue fixation and the number of samples could impact on viral genome detection on EMB.

scholarly journals Complete Coding Sequence of a Chikungunya Virus Strain Imported into Slovenia from Thailand in Late 2018

2019 ◽  
Vol 8 (37) ◽  
Author(s):  
Samo Zakotnik ◽  
Misa Korva ◽  
Nataša Knap ◽  
Barbara Robnik ◽  
Nina Gorišek Miksić ◽  
...  
Keyword(s):  
Cell Culture ◽  
Phylogenetic Analysis ◽  
Envelope Protein ◽  
Viral Genome ◽  
Chikungunya Virus ◽  
Whole Genome ◽  
Reverse Transcription Pcr ◽  
The Indian Ocean ◽  
Complete Viral Genome ◽  
A226v Mutation

A case of chikungunya virus infection was imported from Thailand into Slovenia in late 2018. The infection was diagnosed using real-time reverse transcription-PCR, the virus was isolated in cell culture, and the whole genome was sequenced. Phylogenetic analysis of the nearly complete viral genome indicated that the virus belongs to the Indian Ocean lineage but does not possess the A226V mutation in the envelope protein E1.

scholarly journals DISEASES AND MORTALITY IN FREE-RANGING BROWN BEAR (URSUS ARCTOS), GRAY WOLF (CANIS LUPUS), AND WOLVERINE (GULO GULO) IN SWEDEN

2005 ◽  
Vol 41 (2) ◽  
pp. 298-303 ◽  
Author(s):  
Torsten Mörner ◽  
Hanna Eriksson ◽  
Caroline Bröjer ◽  
Kristina Nilsson ◽  
Henrik Uhlhorn ◽  
...  
Keyword(s):  
Canis Lupus ◽  
Ursus Arctos ◽  
Brown Bear ◽  
Gray Wolf ◽  
Gulo Gulo ◽  
Free Ranging

scholarly journals Treatment of a forelimb fracture and rehabilitation of a free-ranging Iberian Wolf (Canis lupus signatus)

2016 ◽  
Vol 36 (5) ◽  
pp. 412-416 ◽  
Author(s):  
Filipe C. Silva ◽  
Roberto F. Sargo ◽  
Luís C. Sousa ◽  
Helena Rio-Maior ◽  
Ricardo Brandão ◽  
...  
Keyword(s):  
Canis Lupus ◽  
Wound Management ◽  
Long Bone ◽  
Operative Care ◽  
Wildlife Tracking ◽  
Osteosynthesis Plate ◽  
The Right ◽  
Free Ranging ◽  
Canis Lupus Signatus ◽  
Iberian Wolf

Abstract: The surgical treatment of an exposed compounded comminuted fracture of the right radius and ulna in a free-ranging adult female Iberian Wolf (Canis lupus signatus) with an osteosynthesis plate and screws and subsequent post-operative care are described. The evolution of the fracture healing was very similar to those expected in a dog of the same size. The prompt surgical intervention and a proper housing, feeding and wound management adapted to a free-ranging wolf, in view to reduce manipulation and post-operative complications, allowed the subsequent rehabilitation and release of the animal. After 10th post-operative weeks the wolf was fitted with a Global Positioning System (GPS) for wildlife tracking collar and released in the same area where it has been caught. GPS telemetry data showed that the animal covered increasingly large distances confirming a complete functionality of the right thoracic limb and its successfully return to the wild. This report could constitute the first detailed report of a long bone fracture treatment in a free-ranging wolf and its successfully rehabilitation, release and adaptation to the wild.

scholarly journals Hitchhiking of Viral Genomes on Cellular Chromosomes

2019 ◽  
Vol 6 (1) ◽  
pp. 275-296 ◽  
Author(s):  
Tami L. Coursey ◽  
Alison A. McBride
Keyword(s):  
Viral Genome ◽  
Viral Infections ◽  
Genome Replication ◽  
Mitotic Chromosomes ◽  
Dna Viruses ◽  
Viral Genomes ◽  
Barr Virus ◽  
Dividing Cells ◽  
Viral Genome Replication ◽  
Epstein Barr

Persistent viral infections require a host cell reservoir that maintains functional copies of the viral genome. To this end, several DNA viruses maintain their genomes as extrachromosomal DNA minichromosomes in actively dividing cells. These viruses typically encode a viral protein that binds specifically to viral DNA genomes and tethers them to host mitotic chromosomes, thus enabling the viral genomes to hitchhike or piggyback into daughter cells. Viruses that use this tethering mechanism include papillomaviruses and the gammaherpesviruses Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus. This review describes the advantages and consequences of persistent extrachromosomal viral genome replication.

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