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Microbiology Spectrum Mar 2024Bluetongue virus (BTV) is the causative agent of the important livestock disease bluetongue (BT), which is transmitted via Culicoides bites. BT causes severe economic...
Bluetongue virus (BTV) is the causative agent of the important livestock disease bluetongue (BT), which is transmitted via Culicoides bites. BT causes severe economic losses associated with its considerable impact on health and trade of animals. By reverse genetics, we have designed and rescued reporter-expressing recombinant (r)BTV expressing NanoLuc luciferase (NLuc) or Venus fluorescent protein. To generate these viruses, we custom synthesized a modified viral segment 5 encoding NS1 protein with the reporter genes located downstream and linked by the Porcine teschovirus-1 (PTV-1) 2A autoproteolytic cleavage site. Therefore, fluorescent signal or luciferase activity is only detected after virus replication and expression of non-structural proteins. Fluorescence or luminescence signals were detected in cells infected with rBTV/Venus or rBTV/NLuc, respectively. Moreover, the marking of NS2 protein confirmed that reporter genes were only expressed in BTV-infected cells. Growth kinetics of rBTV/NLuc and rBTV/Venus in Vero cells showed replication rates similar to those of wild-type and rBTV. Infectivity studies of these recombinant viruses in IFNAR(-/-) mice showed a higher lethal dose for rBTV/NLuc and rBTV/Venus than for rBTV indicating that viruses expressing the reporter genes are attenuated . Interestingly, luciferase activity was detected in the plasma of viraemic mice infected with rBTV/NLuc. Furthermore, luciferase activity quantitatively correlated with RNAemia levels of infected mice throughout the infection. In addition, we have investigated the replication and dissemination of BTV in IFNAR (-/-) mice using BTV/NLuc and non-invasive imaging systems.IMPORTANCEThe use of replication-competent viruses that encode a traceable fluorescent or luciferase reporter protein has significantly contributed to the and study of viral infections and the development of novel therapeutic approaches. In this work, we have generated rBTV that express fluorescent or luminescence proteins to track BTV infection both and . Despite the availability of vaccines, BTV and other related orbivirus are still associated with a significant impact on animal health and have important economic consequences worldwide. Our studies may contribute to the advance in orbivirus research and pave the way for the rapid development of new treatments, including vaccines.
Topics: Chlorocebus aethiops; Animals; Mice; Bluetongue virus; Genes, Reporter; Vero Cells; Viral Proteins; Luciferases; Vaccines
PubMed: 38353566
DOI: 10.1128/spectrum.02493-23 -
American Journal of Veterinary Research Apr 2024The objective of this study was to determine the seroprevalence of reproductive and infectious diseases in tropical cattle in the Tambopata and Tahuamanu Provinces in...
OBJECTIVE
The objective of this study was to determine the seroprevalence of reproductive and infectious diseases in tropical cattle in the Tambopata and Tahuamanu Provinces in the department of Madre de Dios, Peru.
SAMPLE
156 bovines from 7 cattle farms were sampled. These farms used exclusive grazing for food and natural mating for reproduction and did not have sanitary or vaccination programs.
METHODS
The serum of blood samples was subjected to ELISA with commercial kits for the detection of antibodies against Neospora caninum, Mycobacterium avium subsp paratuberculosis (MAP), Leptospira interrogans, pestivirus bovine viral diarrhea virus-1, retrovirus bovine leukemia virus (BLV), orbivirus bluetongue virus (BTV), and herpesvirus bovine herpes virus-1 (BHV). The data were analyzed by means of association tests with χ2 (P < .05) and Spearman rank correlation (P < .05) in the SPSS v.15.0 software (IBM Corp).
RESULTS
A low prevalence of antibodies to L interrogans, N caninum, M avium subsp paratuberculosis, bovine viral diarrhea virus-1 was found, but it was high to BTV, BLV, and BHV (100%, 53.85%, and 72.44%, respectively). The presence of BLV and BHV was higher in the Las Piedras District, bovines less than 5 years old, and cattle with breed characteristics of zebu and crossbred (P < .01). In addition, there was a significant correlation between both infections, showing 83.3% of BLV positivity that were also BHV positive (P < .01).
CLINICAL RELEVANCE
The high prevalence of antibodies to BTV, BHV, and BLV could be due to livestock management practices, direct contact with infected animals, and variation of the presence of vectors and natural reservoirs in the context of climate change in the tropics.
Topics: Cattle; Animals; Paratuberculosis; Cattle Diseases; Enzootic Bovine Leukosis; Bovine Virus Diarrhea-Mucosal Disease; Peru; Seroepidemiologic Studies; Diarrhea Viruses, Bovine Viral; Antibodies, Viral; Antibodies, Bacterial; Communicable Diseases; Reproduction; Diarrhea Virus 1, Bovine Viral; Leukemia Virus, Bovine; Diarrhea
PubMed: 38335721
DOI: 10.2460/ajvr.23.08.0177 -
Journal of Medical Entomology Mar 2024Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) are arthropod-borne viruses that are transmitted by biting midges in the genus Culicoides (Diptera:...
Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) are arthropod-borne viruses that are transmitted by biting midges in the genus Culicoides (Diptera: Ceratopogonidae) and can cause hemorrhagic disease in certain ruminants. The objectives of this study were to measure the incidence of BTV and EHDV infections in captive white-tailed deer herd as well as tissues and corresponding presence of Culicoides midges at a location near Clinton, LA. During a 7-yr study with yearly outbreaks of hemorrhagic disease in the deer herd, 15 species of Culicoides were captured using Centers for Disease Control (CDC) black light traps. Reverse transcriptase quantitative polymerase chain reaction (PCR) was performed to screen for BTV and EHDV in pools of midges and tissues of deer. From 2012 to 2018, 1,711 pools of midges representing 24,859 specimens were tested, and specimens from 5 of the 15 collected species (Culicoides debilipalpis, Culicoides stellifer, Culicoides venustus, Culicoides haematopotus, and Culicoides crepuscularis) were found to be PCR positive for BTV and EHDV. Most of the BTV-positive pools of biting midges were from specimens of C. debilipalpis and C. stellifer, and most of the EHDV-positive pools were from specimens of C. venustus and C. stellifer. During the 7-yr period, 112 white-tailed deer that died at the study location were PCR positive for BTV or EHDV: detected BTV serotypes were 10 and 12 and EHDV serotypes were 1, 2, and 6. There was a significant increase in BTV/EHDV antibody prevalence in white-tailed deer during the study; antibody-positive rates increased from 15% to 78% in the deer herd of approximately 100 animals.
Topics: Animals; Sheep; Ceratopogonidae; Reoviridae Infections; Deer; Prospective Studies; Incidence; Insect Vectors; Ruminants; Bluetongue; Virus Diseases; Hemorrhagic Disease Virus, Epizootic; Bluetongue virus; Sheep Diseases
PubMed: 38297491
DOI: 10.1093/jme/tjae006 -
Viruses Jan 2024In Cuba, despite a high sero-prevalence of bluetongue virus (BTV), circulating serotypes remain unknown. The aim of this study was to identify circulating BTV serotypes...
In Cuba, despite a high sero-prevalence of bluetongue virus (BTV), circulating serotypes remain unknown. The aim of this study was to identify circulating BTV serotypes in farms throughout the western region of Cuba. Blood samples were collected from 200 young cattle and sheep between May and July 2022 for virological analyses (PCR, viral isolation and virus neutralization) and genome sequencing. The results confirmed viral circulation, with viro-prevalence of 25% for BTV. The virus was isolated from 18 blood samples and twelve BTV serotypes were identified by sequencing RT-PCR products targeting the segment 2 of the BTV genome (BTV-1, 2, 3, 6, 10, 12, 13, 17, 18, 19, 22 and 24). Finally, the full genome sequences of 17 Cuban BTV isolates were recovered using a Sequence Independent Single Primer Amplification (SISPA) approach combined to MinION Oxford Nanopore sequencing technology. All together, these results highlight the co-circulation of a wide diversity of BTV serotypes in a quite restricted area and emphasize the need for entomological and livestock surveillance, particularly in light of recent changes in the global distribution and nature of BTV infections.
Topics: Sheep; Animals; Cattle; Serogroup; Bluetongue; Cuba; Base Sequence; Bluetongue virus
PubMed: 38275974
DOI: 10.3390/v16010164 -
Frontiers in Cellular and Infection... 2023plays a crucial role as an insect vector in the field of veterinary medicine. The transmission of significant viruses such as bluetongue virus (BTV) and African horse...
INTRODUCTION
plays a crucial role as an insect vector in the field of veterinary medicine. The transmission of significant viruses such as bluetongue virus (BTV) and African horse sickness virus (AHSV) by this insect poses a substantial threat, leading to the development of severe diseases in domestic animals. This study aimed to explore the species, identify their blood-meal sources, and assess the presence of BTV and AHSV carried by in Yuanyang County, Yunnan Province. The aim was to gain insights into the potential vectors of these two viruses and elucidate their potential roles in the transmission of pathogens.
METHODS
The midges were collected from cattle (), pig (), and goat () pens in Yuanyang County, Yunnan Province in June 2020. Initial identification of midges was conducted through morphological characteristics, followed by molecular identification using the cytochrome C oxidase subunit I (COI) gene. The determination of blood-meal sources was accomplished using specific primers targeting the cytochrome (Cyt ) gene from potential hosts. BTV and AHSV RNA were identified in pools through the application of reverse transcriptase PCR and quantitative real-time PCR. Nucleotide homology and phylogenetic analysis were performed using MegAlign (DNAStar) and Mega 6.0 software.
RESULTS
A total of 6,300 , consisting of , and , were collected from cattle, pigs, and goat pens. The engorgement rates for these species were 30.2%, 54.6%, 75%, and 66.7%, respectively. In the cattle pen, the prevailing species is (100%). In the pig pen, dominates (70%), with following at 30%. In the goat pen, holds the majority (45.45%), trailed by (25%), (20.45%), and (9.09%). These species were identified as feeding on cattle, pigs, goats, chickens (), and humans (). The positivity rates for BTV were 20.00% and 11.54% in blood-fed specimens of and , respectively. Conversely, the positivity rates for BTV in non-blood-fed specimens were 0.00% and 6.67% for and , respectively. BTV was not detected in and . The specimens (YY86) from that tested positive for BTV had the closest genetic relationship to YTS-4 isolated from Mangshi, Yunnan Province in 1996. All test results for the nucleic acid of AHSV were negative.
CONCLUSION
The study reveals variations in the species distribution, community composition, blood sucking rate, and blood-feeding sources of across different habitats. Notably, and emerge as potential vectors for the transmission of BTV in local animals. Accordingly, this investigation provides crucial insights that can serve as a valuable reference for the prevention and control of BTV in local animals, particularly from the perspective of vector management.
Topics: Sheep; Humans; Cattle; Animals; Ceratopogonidae; Bluetongue virus; Phylogeny; Bluetongue; China; Chickens; Goats
PubMed: 38274733
DOI: 10.3389/fcimb.2023.1283216 -
Journal of Thermal Biology Jan 2024Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are hematophagous insects, and some species can transmit a plethora of pathogens, e.g., bluetongue virus...
Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are hematophagous insects, and some species can transmit a plethora of pathogens, e.g., bluetongue virus and African horse sickness virus, that mainly affect animals. The transmission of vector-borne pathogens is strongly temperature dependent, and recent studies pointed to the importance of including microclimatic data when modelling disease spread. However, little is known about the preferred temperature of biting midges. The present study addressed the thermal selection of field-caught Culicoides with two experiments. In a laboratory setup, sugar-fed or blood-fed midges were video tracked for 15 min while moving inside a 60 × 30 × 4 cm setup with a 15-25 °C temperature gradient. Culicoides spent over double the time in the coldest zone of the setup compared to the warmest one. This cold selection was significantly stronger for sugar-fed individuals. Calculated preferred temperatures were 18.3 °C and 18.9 °C for sugar-fed and blood-fed Culicoides, respectively. The effect of temperature on walking speed was significant but weak, indicating that their skewed distribution results from preference and not cold trapping. A second experiment consisted of a two-way-choice-setup, performed in a 90 × 45 × 45 cm net cage, placed outdoors in a sheltered environment. Two UV LED CDC traps were placed inside the setup, and a mean temperature difference of 2.2 °C was created between the two traps. Hundred-fifty Culicoides were released per experiment. Recapture rates were negatively correlated with ambient temperature and were on average three times higher in the cooled trap. The higher prevalence of biting midges in cooler environments influences fitness and ability to transmit pathogens and should be considered in models that predict Culicoides disease transmission.
Topics: Humans; Animals; Ceratopogonidae; Insect Vectors; African Horse Sickness Virus; Environment; Sugars
PubMed: 38244238
DOI: 10.1016/j.jtherbio.2024.103783 -
Virus Genes Feb 2024Bluetongue disease is a reportable animal disease that affects wild and farmed ruminants, including white-tailed deer (WTD). This report documents the clinical findings,...
A novel bluetongue virus serotype 2 strain isolated from a farmed Florida white-tailed deer (Odocoileus virginianus) arose from reassortment of gene segments derived from co-circulating serotypes in the Southeastern USA.
Bluetongue disease is a reportable animal disease that affects wild and farmed ruminants, including white-tailed deer (WTD). This report documents the clinical findings, ancillary diagnostics, and genomic characterization of a novel reassortant bluetongue virus serotype 2 (BTV-2) strain isolated from a dead Florida farmed WTD in 2022. Our analyses support that this BTV-2 strain likely stemmed from the acquisition of genome segments from co-circulating BTV strains in Florida and Louisiana. In addition, our analyses also indicate that genetically uncharacterized BTV strains may be circulating in the Southeastern USA; however, the identity and reassortant status of these BTV strains cannot be determined based on the VP2 and VP5 genome sequences. Hence, continued surveillance based on complete genome characterization is needed to understand the genetic diversity of BTV strains in this region and the potential threat they may pose to the health of deer and other ruminants.
Topics: Animals; Florida; Deer; Bluetongue virus; Serogroup
PubMed: 38182930
DOI: 10.1007/s11262-023-02047-2 -
Veterinary Microbiology Feb 2024We report the discovery of two bluetongue virus serotype 6 (BTV-6) reassortants recovered from a domestic sheep and a free-ranging mule deer in northern Colorado. At the...
We report the discovery of two bluetongue virus serotype 6 (BTV-6) reassortants recovered from a domestic sheep and a free-ranging mule deer in northern Colorado. At the time of this publication, whole-genome sequencing of BTV-6 isolates in the Western U.S. have not been undertaken. These findings reflect the incursive movement of geographically distinct BTV serotypes into important agricultural areas of the U.S. and demonstrate reassortment with regionally circulating serotypes.
Topics: Sheep; Animals; Sheep, Domestic; Deer; Bluetongue; Serogroup; Bluetongue virus; Colorado; Equidae; Sheep Diseases
PubMed: 38141398
DOI: 10.1016/j.vetmic.2023.109944 -
Veterinaria Italiana Dec 2023Epizootic hemorrhagic disease virus serotype 8 (EHDV-8) emerged in Europe for the first time in late 2022. In this study, we investigated the kinetics of EHDV-8...
Epizootic hemorrhagic disease virus serotype 8 (EHDV-8) emerged in Europe for the first time in late 2022. In this study, we investigated the kinetics of EHDV-8 infection in cattle, sheep, and goats. Following experimental infection with EHDV-8, four out of five calves displayed fever, while another calf exhibited ulcerative and crusty lesions of the muzzle. RNAemia peaked at day 7 post infection in all calves and remained relatively stable till the end of the study, at 78 days post infection. Infectious virus was isolated up to 21 days post infection in one calf. As far as small ruminants are concerned, one sheep experienced fever and two out of five had consistent RNAemia that lasted until the end of the study. Remarkably, infectious virus was evidenced at day 7 post infection in one sheep. In goats, no RNA was observed. All infected animals seroconverted, and a neutralizing immune response was observed in all species, with calves exhibiting a more robust response than sheep and goats. Our study provides insights into the kinetics of EHDV-8 infection and the host immune responses. We also highlight that sheep may also play a role in EHDV-8 epidemiology. Altogether, the data gathered in this study could have important implications for disease control and prevention strategies, providing crucial information to policy makers to mitigate the impact of this viral disease on livestock.
Topics: Sheep; Cattle; Animals; Reoviridae Infections; Goats; Serogroup; Hemorrhagic Disease Virus, Epizootic; Cattle Diseases; Ruminants; Goat Diseases; Sheep Diseases
PubMed: 38117055
DOI: 10.12834/VetIt.3433.23112.1 -
Virology Feb 2024The Ibaraki virus (IBAV) causes Ibaraki disease in cattle. Our previous studies have shown that IBAV uses macropinocytosis to enter the host cell and exit from the...
The Ibaraki virus (IBAV) causes Ibaraki disease in cattle. Our previous studies have shown that IBAV uses macropinocytosis to enter the host cell and exit from the endosome to the cytosol in response to endosomal acidification. To further explore the mechanism of IBAV infection and replication, we examined the effect of inhibitors of mitochondrial oxidative phosphorylation, carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and antimycin A, on IBAV propagation. These inhibitors significantly suppressed IBAV propagation, with reduced cellular ATP levels resulting from suppression of ATP synthesis. Furthermore, we identified AMP-activated protein kinase (AMPK), which is activated by CCCP or antimycin A, as a key signaling molecule in IBAV suppression. We also observed that IBAV infection induces ATP depletion and increases AMPK activity. Our findings suggest that AMPK is a potential target in Ibaraki disease.
Topics: Animals; Cattle; AMP-Activated Protein Kinases; Antimycin A; Carbonyl Cyanide m-Chlorophenyl Hydrazone; Mitochondria; Adenosine Triphosphate
PubMed: 38103268
DOI: 10.1016/j.virol.2023.109943