Evaluation of a real-time two-step RT-PCR assay for quantitation of Chronic bee paralysis virus (CBPV) genome in experimentally-infected bee tissues and in life stages of a symptomatic colony

Blanchard, Philippe; Ribière, Magali; Celle, Olivier; Lallemand, Perrine; Schurr, Frank M.; Olivier, Violaine; Iscache, Anne Laure; Faucon, Jean Paul

doi:10.1016/j.jviromet.2006.11.021

Cited by 86 publications

(85 citation statements)

References 19 publications

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“…The genomic load of 10 10 CBPV RNA copies per feces, evaluated by real-time RT-PCR, was consistent with the viral titer of 10 9 particles per bee fecal deposit determined by electronic microscopy (2) and with the estimated genomic CBPV load per symptomatic paralyzed bee, which was up to 10 13 CBPV copies (18). Moreover, the spread of virus within the hive by paralyzed bees was confirmed by detecting the virus in feces from a symptomatic colony that were absorbed onto paper and in apparently unsoiled pieces of this paper.…”

Section: Discussionsupporting

confidence: 81%

See 1 more Smart Citation

Spread of Infectious Chronic Bee Paralysis Virus by Honeybee ( Apis mellifera L.) Feces

Ribière¹,

Lallemand²,

Iscache³

et al. 2007

Appl Environ Microbiol

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Knowledge of the spreading mechanism of honeybee pathogens within the hive is crucial to our understanding of bee disease dynamics. The aim of this study was to assess the presence of infectious chronic bee paralysis virus (CBPV) in bee excreta and evaluate its possible role as an indirect route of infection. Samples of paralyzed bees were (i) produced by experimental inoculation with purified virus and (ii) collected from hives exhibiting chronic paralysis. CBPV in bee heads or feces (crude or absorbed onto paper) was detected by reverse transcription-PCR. CBPV infectivity was assessed by intrathoracic inoculation of bees with virus extracted from feces and by placement of naive bees in cages previously occupied by contaminated individuals. CBPV RNA was systematically detected in the feces of naturally and experimentally infected bees and on the paper sheets that had been used to cover the floors of units containing bees artificially infected with CBPV or the floor of one naturally infected colony. Both intrathoracic inoculation of bees with virus extracted from feces and placement of bees in contaminated cages provoked overt disease in naive bees, thereby proving that the excreted virus was infectious and that this indirect route of infection could lead to overt chronic paralysis. This is the first experimental confirmation that infectious CBPV particles excreted in the feces of infected bees can infect naive bees and provoke overt disease by mere confinement of naive bees in a soiled environment.

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Section: Discussionsupporting

confidence: 81%

“…A two-step real-time RT-PCR assay based on TaqMan technology using a fluorescent probe (6-carboxyfluorescein-6-carboxytetramethylrhodamine) was performed as previously described (18) to quantify the CBPV genomic load in crude feces and on papers with or without feces.…”

Section: Methodsmentioning

confidence: 99%

Spread of Infectious Chronic Bee Paralysis Virus by Honeybee ( Apis mellifera L.) Feces

Ribière¹,

Lallemand²,

Iscache³

et al. 2007

Appl Environ Microbiol

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“…After serial 10-fold dilutions of strong positive DWV and ABPV fi eld samples used to compare conventional and real-time RT-PCR assay, dilution detection limit of real-time RT-PCR was 100 times lower. This proved the higher sensitivity of real-time RT-PCR as reported by Jamnikar Ciglinečki et al [39] and Blanchard et al [57].…”

Section: Discussionsupporting

confidence: 66%

A survey of deformed wing virus and acute bee paralysis virus in honey bee colonies from serbia using real-time RT-PCR

Simeunović¹,

Stevanović²,

Vidanović³

et al. 2014

Acta Veterinaria

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In this study 55 honey bee colonies from different Serbian regions were monitored for the presence of Deformed Wing Virus (DWV) and Acute Bee Paralysis Virus (ABPV) using TaqMan-based real-time RT-PCR assay. The results revealed the presence of DWV in each sampling location, and ABPV in 10 out of 11 apiaries. High frequency of DWV (76.4%) and ABPV (61.8%) positive samples in asymptomatic colonies can be the consequence of inefficient and postponed Varroa treatment concerning the role of this mite in the transmission and activation of honey bee viruses. The real-time RTPCR technique described in this paper is proved to be the most reliable method for this kind of investigation.

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“…It resulted in a ratio of positive-strand to negative-strand genomes of 2500 : 1 in abdomens and 150 : 1 in heads, suggesting a higher replication of DcPV in heads, and a certain affinity of the virus for nervous tissues (neurotropism). Interestingly, neurotropism has been associated with paralytic symptoms of other picorna-like viruses such as Poliovirus [22,23], Aphid Lethal Paralysis Virus [24] and Chronic bee paralysis virus [25]. Remarkably, recovery of normal behaviour is associated with a significant reduction in virus load (figure 5).…”

Section: Results and Discussion (A) The Bodyguard Behaviour: A Neurolmentioning

confidence: 99%

Who is the puppet master? Replication of a parasitic wasp-associated virus correlates with host behaviour manipulation

et al. 2015

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Many parasites modify their host behaviour to improve their own transmission and survival, but the proximate mechanisms remain poorly understood. An original model consists of the parasitoid Dinocampus coccinellae and its coccinellid host, Coleomegilla maculata; during the behaviour manipulation, the parasitoid is not in contact with its host anymore. We report herein the discovery and characterization of a new RNA virus of the parasitoid (D. coccinellae paralysis virus, DcPV). Using a combination of RT-qPCR and transmission electron microscopy, we demonstrate that DcPV is stored in the oviduct of parasitoid females, replicates in parasitoid larvae and is transmitted to the host during larval development. Next, DcPV replication in the host's nervous tissue induces a severe neuropathy and antiviral immune response that correlate with the paralytic symptoms characterizing the behaviour manipulation. Remarkably, virus clearance correlates with recovery of normal coccinellid behaviour. These results provide evidence that changes in ladybeetle behaviour most likely result from DcPV replication in the cerebral ganglia rather than by manipulation by the parasitoid. This offers stimulating prospects for research on parasitic manipulation by suggesting for the first time that behaviour manipulation could be symbiont-mediated.

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Evaluation of a real-time two-step RT-PCR assay for quantitation of Chronic bee paralysis virus (CBPV) genome in experimentally-infected bee tissues and in life stages of a symptomatic colony

Cited by 86 publications

References 19 publications

Spread of Infectious Chronic Bee Paralysis Virus by Honeybee ( Apis mellifera L.) Feces

Spread of Infectious Chronic Bee Paralysis Virus by Honeybee ( Apis mellifera L.) Feces

A survey of deformed wing virus and acute bee paralysis virus in honey bee colonies from serbia using real-time RT-PCR

Who is the puppet master? Replication of a parasitic wasp-associated virus correlates with host behaviour manipulation

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