scholarly journals A Reverse Genetics System for the Israeli Acute Paralysis Virus and Chronic Bee Paralysis Virus

2020 ◽  
Vol 21 (5) ◽  
pp. 1742 ◽  
Author(s):  
Sa Yang ◽  
Hongxia Zhao ◽  
Yanchun Deng ◽  
Shuai Deng ◽  
Xinling Wang ◽  
...  

Honey bee viruses are associated with honey bee colony decline. Israeli acute paralysis virus (IAPV) is considered to have a strong impact on honey bee survival. Phylogenetic analysis of the viral genomes from several regions of the world showed that various IAPV lineages had substantial differences in virulence. Chronic bee paralysis virus (CBPV), another important honey bee virus, can induce two significantly different symptoms. However, the infection characteristics and pathogenesis of IAPV and CBPV have not been completely elucidated. Here, we constructed infectious clones of IAPV and CBPV using a universal vector to provide a basis for studying their replication and pathogenesis. Infectious IAPV and CBPV were rescued from molecular clones of IAPV and CBPV genomes, respectively, that induced typical paralysis symptoms. The replication levels and expression proteins of IAPV and CBPV in progeny virus production were confirmed by qPCR and Western blot. Our results will allow further dissection of the role of each gene in the context of viral infection while helping to study viral pathogenesis and develop antiviral drugs using reverse genetics systems.

Insects ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 9 ◽  
Author(s):  
Esmaeil Amiri ◽  
Gregory Seddon ◽  
Wendy Zuluaga Smith ◽  
Micheline K. Strand ◽  
David R. Tarpy ◽  
...  

Queen loss or failure is an important cause of honey bee colony loss. A functional queen is essential to a colony, and the queen is predicted to be well protected by worker bees and other mechanisms of social immunity. Nevertheless, several honey bee pathogens (including viruses) can infect queens. Here, we report a series of experiments to test how virus infection influences queen–worker interactions and the consequences for virus transmission. We used Israeli acute paralysis virus (IAPV) as an experimental pathogen because it is relevant to bee health but is not omnipresent. Queens were observed spending 50% of their time with healthy workers, 32% with infected workers, and 18% without interaction. However, the overall bias toward healthy workers was not statistically significant, and there was considerable individual to individual variability. We found that physical contact between infected workers and queens leads to high queen infection in some cases, suggesting that IAPV infections also spread through close bodily contact. Across experiments, queens exhibited lower IAPV titers than surrounding workers. Thus, our results indicate that honey bee queens are better protected by individual and social immunity, but this protection is insufficient to prevent IAPV infections completely.


Insects ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 60
Author(s):  
Shilpi Bhatia ◽  
Saman S. Baral ◽  
Carlos Vega Melendez ◽  
Esmaeil Amiri ◽  
Olav Rueppell

Among numerous viruses that infect honey bees (Apis mellifera), Israeli acute paralysis virus (IAPV) can be linked to severe honey bee health problems. Breeding for virus resistance may improve honey bee health. To evaluate the potential for this approach, we compared the survival of IAPV infection among stocks from the U.S. We complemented the survival analysis with a survey of existing viruses in these stocks and assessing constitutive and induced expression of immune genes. Worker offspring from selected queens in a common apiary were inoculated with IAPV by topical applications after emergence to assess subsequent survival. Differences among stocks were small compared to variation within stocks, indicating the potential for improving honey bee survival of virus infections in all stocks. A positive relation between worker survival and virus load among stocks further suggested that honey bees may be able to adapt to better cope with viruses, while our molecular studies indicate that toll-6 may be related to survival differences among virus-infected worker bees. Together, these findings highlight the importance of viruses in queen breeding operations and provide a promising starting point for the quest to improve honey bee health by selectively breeding stock to be better able to survive virus infections.


Viruses ◽  
2014 ◽  
Vol 6 (5) ◽  
pp. 2012-2027 ◽  
Author(s):  
Chunsheng Hou ◽  
Hadassah Rivkin ◽  
Yossi Slabezki ◽  
Nor Chejanovsky

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Kerstin Seitz ◽  
Katharina Buczolich ◽  
Alžbeta Dikunová ◽  
Pavel Plevka ◽  
Karen Power ◽  
...  

Abstract Among the many diseases compromising the well-being of the honey bee (Apis mellifera) the chronic paralysis syndrome of adult honey bees is one of the best described. The causative agent, chronic bee paralysis virus (CBPV), is a positive sense, single-stranded RNA virus with a segmented genome. Segment 1 encodes three putative open reading frames (ORFs), including the RNA-dependent RNA polymerase and other non-structural protein coding regions. Segment 2 encodes four putative ORFs, which contain the genes of supposed structural proteins. In this study, we established a reverse genetic system for CBPV by molecular cloning of DNA copies of both genome segments. CBPV rescue was studied in imago and honey bee pupae infection models. Virus replication and progeny virus production was only initiated when capped RNAs of both genome segments were injected in honey bees. As injection of these clonal RNAs caused clinical symptoms similar to wild-type CBPV infection, we conclude that the novel molecular clone fulfilled Koch’s postulates. Our virus clone will enable in-depth analysis of CBPV pathogenesis and help to increase knowledge about this important honey bee disease.


Insects ◽  
2020 ◽  
Vol 11 (6) ◽  
pp. 382 ◽  
Author(s):  
Jessica L. Kevill ◽  
Katie Lee ◽  
Michael Goblirsch ◽  
Erin McDermott ◽  
David R. Tarpy ◽  
...  

Throughout a honey bee queen’s lifetime, she is tended to by her worker daughters, who feed and groom her. Such interactions provide possible horizontal transmission routes for pathogens from the workers to the queen, and as such a queen’s pathogen profile may be representative of the workers within a colony. To explore this further, we investigated known honey bee pathogen co-occurrence, as well as pathogen transmission from workers to queens. Queens from 42 colonies were removed from their source hives and exchanged into a second, unrelated foster colony. Worker samples were taken from the source colony on the day of queen exchange and the queens were collected 24 days after introduction. All samples were screened for Nosema spp., Trypanosome spp., acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), Israeli acute paralysis virus (IAPV), Lake Sinai virus (LSV), and deformed wing virus master variants (DWV-A, B, and C) using RT-qPCR. The data show that LSV, Nosema, and DWV-B were the most abundant pathogens in colonies. All workers (n = 42) were LSV-positive, 88% were Nosema-positive, whilst pathogen loads were low (<1 × 106 genome equivalents per pooled worker sample). All queens (n = 39) were negative for both LSV and Nosema. We found no evidence of DWV transmission occurring from worker to queen when comparing queens to foster colonies, despite DWV being present in both queens and workers. Honey bee pathogen presence and diversity in queens cannot be revealed from screening workers, nor were pathogens successfully transmitted to the queen.


Insects ◽  
2019 ◽  
Vol 10 (5) ◽  
pp. 123
Author(s):  
Esmaeil Amiri ◽  
Gregory Seddon ◽  
Wendy Zuluaga Smith ◽  
Micheline K. Strand ◽  
David R. Tarpy ◽  
...  

It has been brought to our attention that one note was missing in the Funding section of our published paper [...]


2015 ◽  
Vol 93 (9) ◽  
pp. 711-720 ◽  
Author(s):  
Sarah Michaud ◽  
Humberto F. Boncristiani ◽  
Joost W. Gouw ◽  
Micheline K. Strand ◽  
Jeffrey Pettis ◽  
...  

Recent declines in honey bee (Apis mellifera L., 1758) populations worldwide have spurred significant research into the impact of pathogens on colony health. The role of the Israeli acute paralysis virus (IAPV) on hive mortality has become of particular concern since being correlated with colony losses. However, the molecular interactions between IAPV and its host remain largely unknown. To investigate changes in host protein expression during IAPV infection, mass-spectrometry-based quantitative proteomics was used to compare IAPV-infected and healthy pupae. Proteins whose expression levels changed significantly during infection were identified and functional analysis was performed to determine host systems and pathways perturbed by IAPV infection. Among the A. mellifera proteins most affected by IAPV, those involving translation and the ubiquitin–proteasome pathway were most highly enriched and future investigation of these pathways will be useful in identifying host proteins required for infection. This analysis represents an important first step towards understanding the honey bee host response to IAPV infection through the systems-level analysis of protein expression.


2018 ◽  
Vol 62 (1) ◽  
pp. 141-144 ◽  
Author(s):  
Mayra C. García-Anaya ◽  
Alejandro Romo-Chacón ◽  
Alma I. Sáenz-Mendoza ◽  
Gerardo Pérez-Ordoñez ◽  
Carlos H. Acosta-Muñiz

Abstract The recent alarming loss of honey bee colonies around the world is believed to be related to the presence of viruses. The aim of this study was to detect two major viral diseases, Apis mellifera Filamentous virus (AmFV) and Israeli Acute Paralysis Virus (IAPV) using Reverse Transcription - Polymerase Chain Reaction RT-PCR, in honey bees in Mexico. Adult and larvae honey bee samples were collected from asymptomatic colonies of six major beekeeping regions in the state of Chihuahua, Mexico. Both viruses were detected in both developmental stages of honey bees, IAPV at a higher prevalence (23.5%) as compared to AmFV, only in 0.9% of samples. However, this is the first report on AmFV infection in Mexican apiaries. Further studies are required to understand the AmFV and IAPV impact on colony loss in Mexico and to develop strategies for enhancing the control of viral diseases.


2019 ◽  
Vol 16 (1) ◽  
Author(s):  
Peng Liu ◽  
Liqin Yang ◽  
Jingyue Zhang ◽  
Tao Wang ◽  
Yuanyuan Wu ◽  
...  

Abstract Background Goose parvovirus (GPV) is the etiological agent of Derzsy’s disease and is fatal for gosling. Research on the molecular basis of GPV pathogenicity has been hampered by the lack of a reliable reverse genetics system. At present, the GPV infectious clone has been rescued by transfection in the goose embryo, but the growth character of it is unclear in vitro. Methods In this study, we identified the full-length genome of GPV RC16 from the clinical sample, which was cloned into the pACYC177, generating the pIRC16. The recombinant virus (rGPV RC16) was rescued by the transfection of pIRC16 into goose embryo fibroblasts (GEFs). The rescued virus was characterized by whole genome sequencing, indirect immunofluorescence assays (IFA) and western blot (WB) using rabbit anti-GPV Rep polyclonal antibody as the primary antibody. Previously, we found the 164 K, 165 K, and 167 K residues in the 160YPVVKKPKLTEE171 are required for the nuclear import of VP1 (Chen S, Liu P, He Y, et al. Virology 519:17–22). According to that, the GPV infectious clones with mutated K164A, K165A, or K167A in VP1 were constructed, rescued and passaged. Results The rGPV RC16 has been successfully rescued by transfection of pIRC16 into the GEFs and can proliferate in vitro. Furthermore, the progeny virus produced by pIRC16 transfected cells was infectious in GEFs. Moreover, mutagenesis experiments showed that the rGPV RC16 with mutated 164 K, 165 K and 167 K in VP1 could not proliferate in GEFs based on the data of IFA and WB in parental virus and progeny virus. Conclusions The rGPV RC16 containing genetic maker and the progeny virus are infectious in GEFs. The 164 K, 165 K, and 167 K of VP1 are vital for the proliferation of rGPV RC16 in vitro.


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