Isolation of Arboviruses From Cattle and Insects at 2 Sentinel Sites in Queensland, Australia, 1979-85

1990 ◽  
Vol 38 (1) ◽  
pp. 25 ◽  
Author(s):  
DH Cybinski ◽  
MJ Muller
Keyword(s):  
Cell Cultures ◽  
Virus Isolation ◽  
Hemorrhagic Disease ◽  
Tissue Cultures ◽  
Akabane Virus ◽  
Biting Midges ◽  
Bovine Ephemeral Fever ◽  
Serotype 1 ◽  
Intrathoracic Inoculation ◽  
First Time

Blood samples were collected regularly from two sentinel herds of cattle in northern and southern Queensland between 1979 and 1985. From 2660 samples, virus isolation attempts using baby hamster kidney (BHK21) and Aedes albopictus (AA) tissue cultures and suckling mice produced 308 viruses of which 243 (79%) were in the Palyam subgroup of orbiviruses. Mosquitoes and biting midges were collected at the southern sentinel herd site in January-February 1984 and processed for virus isolation in BHK2l and AA tissue cultures and by intrathoracic inoculation of Ae. aegypti mosquitoes. Totals of 14 338 midges of four species in 156 pools, and 9030 mosquitoes of 27 species in 232 pools, were processed and yielded 59 isolations. Of the 35 viruses isolated from Culicoides brevitarsis, 17 were members of the Palyam subgroup. Bovine ephemeral fever (BEF) virus was isolated once from Anopheles bancroftii, once from C. brevitarsis and 17 times from cattle. Akabane virus was isolated for the first time from C. wadai, as well as a further 10 times from C. brevitarsis and 20 times from cattle. Other viruses isolated from cattle included bluetongue serotype 1, and serotypes 5, 6 and 7 of epizootic hemorrhagic disease of deer (EHD). A new BEF group virus, tentatively called Oak Vale, was isolated nine times from Culex edwardsi mosquitoes. Of the orbiviruses, those in the Palyam subgroup were isolated almost exclusively in BHK2l tissue cultures but those in the bluetongue and EHD subgroups were isolated almost exclusively in AA cell cultures or after passage through Ae. aegypti. Of 22 rhabdovirus isolations from blood and insects (BEF, Kimberley and Tibrogargan), 16 were made only in AA cell cultures or after passage through Ae. aegypti.

scholarly journals Bovine ephemeral fever epidemics in Kingdom Saudi Arabia: clinical, epidemiological and molecular investigation

2017 ◽  
Vol 11 (11) ◽  
pp. 854-860 ◽  
Author(s):  
Ahmed A Zaghawa ◽  
Fadhel Housawi ◽  
Abdulmohsen Al-Naeem ◽  
Ahmed Elsify ◽  
Yamen Mohammed Hegazy
Keyword(s):  
Saudi Arabia ◽  
Water Buffalo ◽  
Virus Isolation ◽  
Clinical Symptoms ◽  
Rt Pcr ◽  
Serum Samples ◽  
Bovine Ephemeral Fever ◽  
Geographical Regions ◽  
Epidemiological Pattern ◽  
First Time

Introduction: Bovine ephemeral fever virus (BEFV) is an arthropod borne Rhabdovirus affects cattle and water buffalo causes acute febrile disease. Methodology: The clinical picture and epidemiological pattern of BEF were described among cattle in epidemics of 2007, 2009 and 2011 in four geographical regions of Kingdom Saudi Arabia (Eastern, Jizan, Qasim, and Riyadh). Serum samples were tested using VNT. Virus isolation and molecular characterization were carried out for the first time in KSA. Results: The main clinical symptoms were fever, stiffness, lameness, salivation and subcutaneous emphysema. The prevalence and the mortality rate of BEF have decreased from 70% and 4.6% in 2007 to 30% and 0.6% in 2011, respectively in the 4 studied areas. There was no region association with higher prevalence of BEF. The intracluster correlation (ICC) was estimated for the first time in KSA as 0.0034. BEFV had been isolated from 11 out of 20 samples (55%) and isolation was confirmed by VNT. The molecular detection of BEFV by RT-PCR and real- time RT-qPCR were found more sensitive for diagnosis of the disease than virus isolation; 80% and 90% for the former tests and 55% for the latter. Three isolates were sequenced, they showed 84.7% - 100% identities in between and shared 90.4%-96.5% sequence identity with a previously published sequence from Australia (KF679404). The generated sequences belonged to 3rd cluster of BEFV glycoprotein. Conclusions: BEF occurrence has cyclic nature and the efficacy of vaccines prepared from local strains has to be evaluated and considered in diseases control.

scholarly journals A Study on the Identification of Five Arboviruses from Hematophagous Mosquitoes and Midges Captured in Some Parts of Northern Turkey

2019 ◽  
Author(s):  
Emre Ozan ◽  
Harun Albayrak ◽  
Semra Gumusova ◽  
Cenk S. Bolukbas ◽  
Mithat Kurt ◽  
...  
Keyword(s):  
Climate Changes ◽  
The Black Sea ◽  
Morphological Identification ◽  
Hemorrhagic Disease ◽  
Bovine Ephemeral Fever Virus ◽  
Biting Midges ◽  
Black Sea Region ◽  
Blood Sucking ◽  
Bovine Ephemeral Fever ◽  
Natural Streams

Background: Whether zoonotic or not, arboviral infections are continuing to be a major threat to human health as well as the livestock industry all around the world. This project presented the results of the identification study on five arboviruses, including West Nile virus (WNV), Bovine ephemeral fever virus, Akabane virus, Bluetongue virus, and Epizootic hemorrhagic disease virus, in mosquitos and midges from eight provinces of the Black Sea Region. Methods: During 2011 and 2012, 3193 mosquitoes were captured around natural streams, rivers, lakes, and ponds using dry-baited miniature light-traps. Identification studies were concluded by employing molecular methods. Results: According to the morphological identification, blood-sucking mosquitoes and biting-midges belonged to Aedes (44.69%), Anopheles (28.34%), Culex (22.14%) and Culicoides (4.83%) species. Overall, 146 pools were made up of captured mosquitos and midges. None of the five viruses were directly identified by mosquitoes. Conclusion: Mosquitoes and midges have got a crucial role in the transmission of arboviruses. The risk of occurrence for the investigated arboviruses will continue depending upon many factors including the presence of these viruses in Turkey and its neighboring countries, uncontrolled livestock movements, global warming and climate changes.

scholarly journals Susceptibility of Midge and Mosquito Vectors to SARS-CoV-2

2021 ◽  
Author(s):  
Velmurugan Balaraman ◽  
Barbara S Drolet ◽  
Natasha N Gaudreault ◽  
William C Wilson ◽  
Jeana Owens ◽  
...  
Keyword(s):  
Cell Lines ◽  
Potential Role ◽  
Animal Origin ◽  
Culex Tarsalis ◽  
Biting Midges ◽  
Bacterial Diseases ◽  
Culicoides Sonorensis ◽  
Insect Cell Lines ◽  
Intrathoracic Inoculation

Abstract SARS-CoV-2 is a recently emerged, highly contagious virus and the cause of the current COVID-19 pandemic. It is a zoonotic virus, although its animal origin is not clear yet. Person-to-person transmission occurs by inhalation of infected droplets and aerosols, or by direct contact with contaminated fomites. Arthropods transmit numerous viral, parasitic, and bacterial diseases; however, the potential role of arthropods in SARS-CoV-2 transmission is not fully understood. Thus far, a few studies have demonstrated that SARS-CoV-2 replication is not supported in cells from certain insect species nor in certain species of mosquitoes after intrathoracic inoculation. In this study, we expanded the work of SARS-CoV-2 susceptibility to biting insects after ingesting a SARS-CoV-2-infected bloodmeal. Species tested included Culicoides sonorensis (Wirth & Jones) (Diptera: Ceratopogonidae) biting midges, as well as Culex tarsalis (Coquillett) and Culex quinquefasciatus (Say) mosquitoes (Diptera: Culicidae), all known biological vectors for numerous RNA viruses. Arthropods were allowed to feed on SARS-CoV-2-spiked blood and at a time point postinfection analyzed for the presence of viral RNA and infectious virus. Additionally, cell lines derived from C. sonorensis (W8a), Aedes aegypti (Linnaeus) (Diptera: Culicidae) (C6/36), Cx. quinquefasciatus (HSU), and Cx. tarsalis (CxTrR2) were tested for SARS-CoV-2 susceptibility. Our results indicate that none of the biting insects, nor the insect cell lines evaluated support SARS-CoV-2 replication, suggesting that these species are unable to be biological vectors of SARS-CoV-2.

scholarly journals Isolation of Porcine Circovirus-like Viruses from Pigs with a Wasting Disease in the USA and Europe

1998 ◽  
Vol 10 (1) ◽  
pp. 3-10 ◽  
Author(s):  
G. M. Allan ◽  
F. McNeilly ◽  
S. Kennedy ◽  
B. Daft ◽  
J. A. Ellis ◽  
...  
Keyword(s):  
Cell Culture ◽  
Lymph Node ◽  
Nucleic Acid ◽  
Cell Cultures ◽  
Virus Isolation ◽  
Chicken Anemia Virus ◽  
Porcine Circovirus ◽  
Tissue Homogenate ◽  
Mesenteric Lymph ◽  
Viral Particles

Samples of lung, liver, kidney, pancreas, spleen, and lymph node from pigs with postweaning multisystemic wasting syndrome from California (USA) and samples of mesenteric lymph nodes from similarly diseased pigs from Brittany (France) were examined by light microscopy, in situ hybridization (ISH), and/or virus isolation. Whole genomic probes for porcine circovirus (PCV) and chicken anemia virus (CAV) were used for ISH. Tissue homogenate supernatants were inoculated onto PK/15 cells for virus isolation, and the presence of viral antigen and viral particles was verified by indirect immunofluorescence, ISH, and electron microscopy. Histologic examination of lung from pigs from California revealed interstitial pneumonia, alveolar epithelial hyperplasia, and basophilic nuclear and cytoplasmic inclusions in mononuclear cell infiltrates and various pulmonary epithelial cells. Granulomatous lymphadenitis with syncytial cells typified the lesions seen in the pigs from France. PCV-like nucleic acid was detected by ISH in lung, pancreas, lymph node, kidney, and liver in pigs from California. Positive signal was also obtained in lymph node sections from pigs from France. Probes for CAV were consistently negative. PK/15 cell cultures inoculated with lung preparations from diseased California pigs and mesenteric lymph node preparations from pigs from France had positive fluorescence by indirect staining for PCV using pooled polyclonal pig sera and hyperimmune rabbit serum and had variable staining with a panel of 7 monoclonal antibodies specific for cell culture contaminant PCV. PCV-like nucleic acid was also detected by ISH in cell cultures. Cytopathic effect was not observed Electron microscopic examination of inoculated cell cultures revealed 17-nm viral particles morphologically consistent with PCV No other virus particles were observed. Although genomic analysis for the definitive identification of these viral isolates remains to be done, the evidence provided strongly suggests that these tissue isolates are closely related to, although antigenically distinct from, the original PCV cell culture contaminant.

scholarly journals RS virus diagnosis: comparison of isolation, immunofluorescence and enzyme immunoassay

1986 ◽  
Vol 81 (2) ◽  
pp. 225-232 ◽  
Author(s):  
Marilda M. Siqueira ◽  
Vanja Ferreira ◽  
Jussara P. Nascimento
Keyword(s):  
Tissue Culture ◽  
Respiratory Syncytial Virus ◽  
Enzyme Immunoassay ◽  
Bacterial Contamination ◽  
Virus Isolation ◽  
Rapid Diagnosis ◽  
Tissue Cultures ◽  
Virus Diagnosis ◽  
Under Five ◽  
Syncytial Virus

Two techniques for rapid diagnosis, immunofluorescence (IFAT) and enzyme immunoassay (EIA), have been compared with virus isolaion in tissue culture for the detection of respiratory syncytial virus (RSV) in specimens of nasopharyngeal secretions. The specimens were obtained from children under five years of age suffering from acute respiratory iliness, during a period of six months from January to June 1982. Of 471 specimens examined 54 (11.5%) were positive by virus isolation and 180 (38.2%) were positive by immunofluorescence. The bacterial contamination of inoculated tissue cultures unfortunately prevented the isolation of virus from many samples. Specimens from 216 children were tested to compare enzyme immunoassay and immunofluorescence. Of these 60 (27%) were positive by EIA and 121 (56%) were positive by IFAT. Our results suggest that the EIA technique although highly specific is rather insensitive. This may be because by the time these tests were done the originl nasopharyngeal secretions were considerably diluted and contained more mucus fragments than the call suspension used for IFAT. Of the three techniques, IFAT gives the best results although EIA may be useful where IFAT is not possible.

scholarly journals Recombinant cDNA Probe from Bluetongue Virus Genome Segment 10 for Identification of Bluetongue Virus

1989 ◽  
Vol 1 (3) ◽  
pp. 237-241 ◽  
Author(s):  
Carlos Alberto de Mattos ◽  
Cecilia Cristina de Mattos ◽  
Bennie Irve Osburn
Keyword(s):  
Bluetongue Virus ◽  
Cdna Probe ◽  
Prototype Strain ◽  
Virus Genome ◽  
Genome Segment ◽  
Hemorrhagic Disease ◽  
Cross Hybridization ◽  
Double Stranded Rna ◽  
Field Isolates ◽  
Serotype 1

Genome segment 10 of bluetongue virus (BTV) serotype 11 UC8 strain was cloned and subsequently hybridized to viral double-stranded RNA extracted from 90 field isolates of BTV serotypes 10, 11, 13, and 17; the prototype strains of BTV 2, 10, 11, 13, and 17; the prototype strain epizootic hemorrhagic disease virus (EHDV) serotype 1; and 4 field isolates of EHDV serotype 2. The 90 field isolates were obtained from different counties in California, Louisiana, and Idaho during the years 1979, 1980, and 1981. The cloned genetic probe hybridized with all the BTV samples tested, showing different degrees of cross-hybridization at the stringency conditions used in this study. This indicated that BTV genome segment 10 has conserved nucleotide sequences among the BTV serotypes 2, 10, 11, 13, and 17. No cross-hybridization signals were detected between the cloned genome segment 10 of BTV 11 UC8 strain and the prototype strain of EHDV serotype 1 and the field isolates of serotype 2. This probe recognized a wide variety of BTV isolates.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Antibody to Epizootic Hemorrhagic Disease of Deer Virus

2000 ◽  
Vol 12 (2) ◽  
pp. 142-145 ◽  
Author(s):  
James O. Mecham ◽  
Michael M. Jochim
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hemorrhagic Disease ◽  
Structural Protein ◽  
Serum Samples ◽  
Epizootic Hemorrhagic Disease ◽  
Diagnostic Potential ◽  
The Us ◽  
Serotype 1 ◽  
Infected Cells

An enzyme-linked immunosorbent assay has been developed to detect antibodies to epizootic hemorrhagic disease of deer virus (EHDV). The assay incorporates a monoclonal antibody to EHDV serotype 2 (EHDV-2) that demonstrates specificity for the viral structural protein, VP7. The assay was evaluated with sequential sera collected from cattle experimentally infected with EHDV serotype 1 (EHDV-1) and EHDV-2, as well as the four serotypes of bluetongue virus (BTV), BTV-10, BTV-11, BTV-13, and BTV-17, that currently circulate in the US. A competitive and a blocking format as well as the use of antigen produced from both EHDV-1-and EHDV-2-infected cells were evaluated. The assay was able to detect specific antibody as early as 7 days after infection and could differentiate animals experimentally infected with EHDV from those experimentally infected with BTV. The diagnostic potential of this assay was demonstrated with field-collected serum samples from cattle, deer, and buffalo.

scholarly journals Retention of Lumpy Skin Disease Virus in Stomoxys Spp (Stomoxys Calsitrans, Stomoxys Sitiens, Stomoxys Indica) following intrathoracic inoculation, Diptera: Muscidae

2020 ◽  
Author(s):  
Arman Issimov ◽  
Lespek Kutumbetov ◽  
Assylbek Zhanabayev ◽  
Nurlybay Kazhgaliyev ◽  
Birzhan Nurgaliyev ◽  
...  
Keyword(s):  
Disease Virus ◽  
Skin Disease ◽  
Virus Isolation ◽  
Emerging Disease ◽  
Lumpy Skin Disease ◽  
Lumpy Skin Disease Virus ◽  
Laboratory Conditions ◽  
Means Of Transmission ◽  
Intrathoracic Inoculation

AbstractLumpy skin disease (LSD) is an emerging disease in cattle in Kazakhstan and the means of transmission remains uncertain. In the current study, acquisition of Lumpy Skin Disease Virus (LSDV) by Stomoxys species following intrathoracic inoculation was demonstrated under laboratory conditions. Flies were injected with a virulent LSDV strain into the thorax region to bypass the midgut barrier. The fate of pathogen in the hemolymph of the flies was further examined using PCR and Virus isolation tests. LSDV was isolated from all three Stomoxys species immediately and up to 24h post intrathoracic inoculation while virus DNA was detectable up to 7d post intrathoracic inoculation.

scholarly journals Capripoxviruses: Exploring the genetic relatedness between field and vaccine strains from Egypt

2019 ◽  
Vol 12 (12) ◽  
pp. 1924-1930
Author(s):  
Sherin Reda Rouby ◽  
Abdel-Hamid Bazid ◽  
Momtaz Wasfy ◽  
Magdy El-Sayed
Keyword(s):  
Cell Cultures ◽  
Vaccine Strain ◽  
Virus Isolation ◽  
Genetic Relatedness ◽  
Phylogenetic Analyses ◽  
Heart Cell ◽  
Fetal Kidney ◽  
Field Isolates ◽  
Gpcr Gene ◽  
Ovine Fetal

Background and Aim: Lumpy skin disease (LSD) and sheep pox are economically important Capripoxvirus-induced diseases of cattle and sheep, respectively. Despite the extensive vaccination program adopted by Egyptian veterinary authorities, LSD and sheep pox are still prevalent and spread throughout the whole country. The current study was designed for molecular characterization and phylogenetic analysis of LSD virus (LSDV) and Sheep pox virus (SPPV) recovered from field cases in Egypt along with vaccinal strains to assess their genetic relatedness. Materials and Methods: Skin biopsies were collected from naturally infected cases of LSD in Ismailia (n=3 farms) and Beni-Suef (n=2 farms) Governorates and sheep pox in Beni-Suef (n=1 flock). Virus isolation was carried out on primary ovine fetal kidney and heart cell cultures. DNA was extracted from infected materials (skin lesions, infected cell cultures) as well as LSDV Neethling vaccine strain and Romanian SPPV vaccine strain. Polymerase chain reaction was performed using oligonucleotide primers targeting the entire open reading frame of G protein-coupled receptors (GPCR) gene and gene sequences were analyzed. Results: Virus isolation on primary ovine fetal kidney and heart cell culture revealed a cytopathic effect at the third passage characterized by rounding of infected cells and margination of nuclear chromatin. Comparative sequence analysis of GPCR gene revealed that Egyptian LSDV isolated from Ismailia and Beni-Suef shared 99:100% nucleotide and amino acid (AA) identities with each other. In comparison to the vaccinal strains, Egyptian LSDV isolates shared 98:99 nucleotide and AA identities with LSDV Neethling vaccine strain and 93:94% with SPPV Romanian vaccine strain. No differences at the nucleotide or AAs were observed between the SPPV vaccine and virulent strains (100% identity). Phylogenetic analyses revealed that LSDV Neethling vaccine strain is more related to field Egyptian LSDV and clustered within the LSDV group while Romanian SPPV vaccine strain clustered in a separate clade with SPPV field isolates. Conclusion: Comparative sequencing and phylogenetic analyses of the GPCR gene reveal a minimal genetic variation between LSDV field isolates from different locations and a close relationship between virulent field strains and homologous vaccines.

New chironomids from Eocene Sakhalinian amber (Diptera; Chironomidae; Orthocladiinae)

2013 ◽  
Vol 6 (1-2) ◽  
pp. 61-69 ◽  
Author(s):  
Viktor Baranov ◽  
Evgeny E. Perkovsky
Keyword(s):  
Baltic Amber ◽  
Biting Midges ◽  
Extant Genus ◽  
The Baltic ◽  
First Time ◽  
Fossil Resins

Non-biting midges (Diptera: Chironomidae) are recorded in the Sakhalinian amber (Russia) for the first time.Pseudorthocladius zherikhinisp. n. is described in an extant genus of Orthocladiinae also known from the Baltic amber.Antillocladussp. (Orthocladiinae) is the first representative of this genus recorded from fossil resins.

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