evolutionary and taxonomic implications of conserved structural motifs between picornaviruses and insect picorna-like viruses.a comparison of the recently determined structure of an insect picorna-like virus, cricket paralysis virus (crpv), with that of the mammalian picornaviruses shows that several structural features are highly conserved between these viruses. these conserved features include the topology of the coat proteins, the conformation of most loops, and the general arrangement of the internally located n-terminal arms of the coat proteins. the conformational conservation of the n-termini of the three major ...200211855636
the complete genome sequence of perina nuda picorna-like virus, an insect-infecting rna virus with a genome organization similar to that of the mammalian picornaviruses.perina nuda picorna-like virus (pnpv) is an insect-infecting rna virus with morphological and physicochemical characters similar to the picornaviridae. in this article, we determine the complete genome sequence and analyze the gene organization of pnpv. the genome of pnpv consists of 9476 nucleotides (nts) excluding the poly(a) tail and contains a single large open reading frame (orf) of 8958 nts (2986 codons) flanked by 473 and 45 nt noncoding regions on the 5' and 3' ends, respectively. northe ...200212009873
novel insect picorna-like virus identified in the brains of aggressive worker identify candidate genes involved in the aggressive behavior of worker honeybees, we used the differential display method to search for rnas exclusively detected in the brains of aggressive workers that had attacked a hornet. we identified a novel, 10,152-nucleotide rna, termed kakugo rna. kakugo rna encodes a protein of 2,893 amino acid residues that shares structural features and sequence similarities with various picorna-like virus polyproteins, especially those from sacbrood virus, which ...200414722264
sequence analysis and genomic organization of a new insect picorna-like virus, ectropis obliqua picorna-like virus, isolated from ectropis obliqua.the complete nucleotide sequence of a new insect picorna-like virus, ectropis obliqua picorna-like virus (eopv), which causes a fatal infection of ectropis obliqua larvae, has been determined. the genomic rna of eopv is 9394 nt in length and contains a single, large open reading frame (nt 391-9351) encoding a polyprotein of 2987 aa. sequence comparisons with other viral polyproteins revealed that the consensus sequences for picornavirus rna helicase, protease and rna-dependent rna polymerase pro ...200415105531
prevalence and seasonal variations of six bee viruses in apis mellifera l. and varroa destructor mite populations in france.a survey of six bee viruses on a large geographic scale was undertaken by using seemingly healthy bee colonies and the pcr technique. samples of adult bees and pupae were collected from 36 apiaries in the spring, summer, and autumn during 2002. varroa destructor samples were collected at the end of summer following acaricide treatment. in adult bees, during the year deformed wing virus (dwv) was found at least once in 97% of the apiaries, sacbrood virus (sbv) was found in 86% of the apiaries, ch ...200415574916
multiple virus infections in the honey bee and genome divergence of honey bee viruses.using uniplex rt-pcr we screened honey bee colonies for the presence of several bee viruses, including black queen cell virus (bqcv), deformed wing virus (dwv), kashmir bee virus (kbv), and sacbrood virus (sbv), and described the detection of mixed virus infections in bees from these colonies. we report for the first time that individual bees can harbor four viruses simultaneously. we also developed a multiplex rt-pcr assay for the simultaneous detection of multiple bee viruses. the feasibility ...200415579317
intricate transmission routes and interactions between picorna-like viruses (kashmir bee virus and sacbrood virus) with the honeybee host and the parasitic varroa mite.viral diseases of honeybees are a major problem in apiculture, causing serious economic losses worldwide, especially in combination with varroa mites. to increase understanding of the relationship among viruses, mites and colony decline, the tripartite relationships among bees, viruses [kashmir bee virus (kbv) and sacbrood virus (sbv)] and varroa mites have been investigated systematically. to develop an antibody-based test for kbv, two structural recombinant proteins were purified for polyclona ...200516033976
detection of multiple viruses in queens of the honey bee apis mellifera l.individual honey bee apis mellifera l. queens were examined for the presence of six honey bee viruses including acute bee paralysis virus, chronic bee paralysis virus, black queen cell virus, deformed wing virus, kashmir bee virus, and sacbrood virus. all viruses, except abpv, were detected in the samples. among queens examined for virus infections, 93% had multiple virus infections. the detection of viruses in queens raises the possibility of a vertical transmission pathway wherein infected que ...200516214161
occurrence of six honeybee viruses in diseased austrian apiaries.the occurrence, prevalence, and distribution patterns of acute bee paralysis virus (abpv), black queen cell virus (bqcv), chronic bee paralysis virus (cbpv), deformed wing virus (dwv), kashmir bee virus (kbv), and sacbrood virus (sbv) were investigated in 90 austrian honeybee colonies suffering from symptoms of depopulation, sudden collapse, paralysis, or dark coloring by employing reverse transcription-pcr. infestation with parasites was also recorded. the samples originated from all parts of a ...200616597939
survey of six bee viruses using rt-pcr in northern thailand.six honey bee viruses were surveyed using rt-pcr in northern thailand where about 80% of thai apiaries are located. tested samples were found to be positive for deformed wing virus (dwv), acute bee paralysis virus (abpv), sacbrood virus (sbv) and kashmir bee virus (kbv). in the collected samples, neither chronic bee paralysis virus nor black queen cell virus nucleic acids could be detected. it was found that dwv was the most widespread and abpv was the second most prevalent. kashmir bee virus wa ...200919105966
presence and prevalence of viruses in local and migratory honeybees (apis mellifera) in massachusetts.migratory and local bees in massachusetts were analyzed for seven viruses. three were detected: black queen cell virus (bqcv), deformed wing virus (dwv), and sacbrood virus (sbv). dwv was most common, followed closely by bqcv and then by sbv. bqcv and sbv were present at significantly higher rates in the migratory bees assayed, bringing into question the impact that these bees have on the health of local bee populations.200919854916
detection of honey bee (apis mellifera) viruses with an oligonucleotide recent years, declines in honey bee (apis mellifera l.) colonies have been observed to varying degrees worldwide with the worst losses in the usa being termed colony collapse disorder (ccd). pathogen load and the prevalence of honey bee viruses have been implicated in these losses and many diseased hives have multiple viruses present. we have designed and tested an oligonucleotide microarray which enables the simultaneous detection of nine honey bee viruses: acute bee paralysis virus, black q ...201121419132
molecular and biological characterization of chinese sacbrood virus ln isolate.chinese sacbrood virus (csbv) was purified from diseased insects, and its genome was cloned and sequenced. the genomic rna of csbv is 8863 nucleotides in length and contains a single large open reading frame encoding a 319.614 kda polyprotein. the coding sequence is flanked by a 178-nucleotide 5' nontranslated leader sequence and a 142-nucleotide 3' nontranslated region, followed a poly(a) tail. four major structural proteins, vp1,vp2, vp3 and vp4, were predicted in the n-teminal of the polyprot ...201121527980
infestation of japanese native honey bees by tracheal mite and virus from non-native european honey bees in japan.invasion of alien species has been shown to cause detrimental effects on habitats of native species. insect pollinators represent such examples; the introduction of commercial bumble bee species for crop pollination has resulted in competition for an ecological niche with native species, genetic disturbance caused by mating with native species, and pathogen spillover to native species. the european honey bee, apis mellifera, was first introduced into japan for apiculture in 1877, and queen bees ...201121960435
Occurrence and prevalence of seven bee viruses in Apis mellifera and Apis cerana apiaries in China.Populations of Apis mellifera and Apis cerana in China were surveyed for seven bee viruses: acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV), Kashmir bee virus (KBV), sacbrood virus (SBV), and Isreal acute paralysis virus (IAPV). No KBV was detected from any samples of the two species. In A. mellifera, DWV was the most prevalent virus, but in A. cerana, SBV was the dominant. Simultaneous multiple infections of viruses ...201222062807
prevalence of nosema and virus in honey bee (apis mellifera l.) colonies on flowering period of acacia in korea.honey production from approximately 1.6 million colonies owned by about 199,000 korean beekeepers was almost 23,000 metric tons in 2009. nosema causes significant losses in honey production and the virus decreases population size. we initiated a survey of honey bee colonies on the blooming period of acacia to determine the prevalence of nosema and virus in 2011. most korean beekeepers have moved from the south to north of korea to get acacia nectar for 2 mon. this provided a valuable opportunity ...201122783125
the iflaviruses sacbrood virus and deformed wing virus evoke different transcriptional responses in the honeybee which may facilitate their horizontal or vertical transmission.sacbrood virus (sbv) and deformed wing virus (dwv) are evolutionarily related positive-strand rna viruses, members of the iflavirus group. they both infect the honeybee apis mellifera but have strikingly different levels of virulence when transmitted orally. honeybee larvae orally infected with sbv usually accumulate high levels of the virus, which halts larval development and causes insect death. in contrast, oral dwv infection at the larval stage usually causes asymptomatic infection with low ...201626819848
the honey bee pathosphere of mongolia: european viruses in central asia.parasites and pathogens are apparent key factors for the detrimental health of managed european honey bee subspecies, apis mellifera. apicultural trade is arguably the main factor for the almost global distribution of most honey bee diseases, thereby increasing chances for multiple infestations/infections of regions, apiaries, colonies and even individual bees. this imposes difficulties to evaluate the effects of pathogens in isolation, thereby creating demand to survey remote areas. here, we co ...201626959221
occurrence of deformed wing virus, chronic bee paralysis virus and mtdna variants in haplotype k of varroa destructor mites in syrian apiaries.a small-scale survey was conducted on 64 beehives located in four governorates of syria in order to assess for the first time the presence of honeybee-infecting viruses and of varroa destructor mites in the country. rt-pcr assays conducted on 192 honeybees (apis mellifera l.) using virus-specific primers showed that deformed wing virus (dwv) was present in 49 (25.5%) of the tested samples and chronic bee paralysis virus (cbpv) in 2 (1.04%), whereas acute bee paralysis virus, sacbrood virus, blac ...201626914360
prevalence of honeybee viruses in different regions of china and argentina.honeybees are threatened by various pathogens and parasites. more than 18 viruses have been described in honeybees and many of them have been detected in china and argentina. in china, both apis cerana and apis mellifera are raised. in argentina, beekeepers raise different ecotypes of a. mellifera: european honeybees (in both temperate and subtropical regions) and africanised honeybees (in subtropical areas only). a thorough study was carried out in both china and argentina to analyse the curren ...201628332647
virus infections of honeybees apis mellifera.the health and vigour of honeybee colonies are threatened by numerous parasites (such as varroa destructor and nosema spp.) and pathogens, including viruses, bacteria, protozoa. among honeybee pathogens, viruses are one of the major threats to the health and well-being of honeybees and cause serious concern for researchers and beekeepers. to tone down the threats posed by these invasive organisms, a better understanding of bee viral infections will be of crucial importance in developing effectiv ...201527800411
bee pathogens found in bombus atratus from colombia: a case study.bombus atratus bumblebees from colombia that were caught in the wild and from breeding programs were screened for a broad set of bee pathogens. we discovered for the first time lake sinai virus and confirmed the infection by other common viruses. the prevalence of apicystis bombi, crithidia bombi and nosema ceranae was remarkably high. according to other studies the former two could have been co-introduced in south america with exotic bumble bees as bombus terrestris or bombus ruderatus. given t ...201526031564
vertical transmission of honey bee viruses in a belgian queen breeding program.the member states of european union are encouraged to improve the general conditions for the production and marketing of apicultural products. in belgium, programmes on the restocking of honey bee hives have run for many years. overall, the success ratio of this queen breeding programme has been only around 50%. to tackle this low efficacy, we organized sanitary controls of the breeding queens in 2012 and 2014.201525889959
lower virus infections in varroa destructor-infested and uninfested brood and adult honey bees (apis mellifera) of a low mite population growth colony compared to a high mite population growth colony.a comparison was made of the prevalence and relative quantification of deformed wing virus (dwv), israeli acute paralysis virus (iapv), black queen cell virus (bqcv), kashmir bee virus (kbv), acute bee paralysis virus (abpv) and sac brood virus (sbv) in brood and adult honey bees (apis mellifera) from colonies selected for high (hmp) and low (lmp) varroa destructor mite population growth. two viruses, abpv and sbv, were never detected. for adults without mite infestation, dwv, iapv, bqcv and kbv ...201525723540
molecular and phylogenetic characterization of honey bee viruses, nosema microsporidia, protozoan parasites, and parasitic mites in china.china has the largest number of managed honey bee colonies, which produce the highest quantity of honey and royal jelly in the world; however, the presence of honey bee pathogens and parasites has never been rigorously identified in chinese apiaries. we thus conducted a molecular survey of honey bee rna viruses, nosema microsporidia, protozoan parasites, and tracheal mites associated with nonnative apis mellifera ligustica and native apis cerana cerana colonies in china. we found the presence of ...201323467539
prevalence and distribution of six bee viruses in korean apis cerana populations.the prevalence and distribution of six bee viruses was investigated in 527 apis cerana samples which were collected from five provinces in south korea. the most prevalent virus, black queen cell virus (bqcv), was present in 75.11% of 446 adult bee samples, followed by sacbrood virus (sbv) in 30.71%. deformed wing virus (dwv), kashmir bee virus (kbv), and chronic bee paralysis virus (cbpv) were present at lower levels of 8.07%, 1.56%, and 0.44%, respectively. the most prevalent virus in 81 larvae ...201222273697
unbiased rna shotgun metagenomics in social and solitary wild bees detects associations with eukaryote parasites and new viruses.the diversity of eukaryote organisms and viruses associated with wild bees remains poorly characterized in contrast to the well-documented pathosphere of the western honey bee, apis mellifera. using a deliberate rna shotgun metagenomic sequencing strategy in combination with a dedicated bioinformatics workflow, we identified the (micro-)organisms and viruses associated with two bumble bee hosts, bombus terrestris and bombus pascuorum, and two solitary bee hosts, osmia cornuta and andrena vaga. i ...201628006002
large pathogen screening reveals first report of megaselia scalaris (diptera: phoridae) parasitizing apis mellifera intermissa (hymenoptera: apidae).as it is most likely that global warming will also lead to a shift in pollinator-habitats northwards, the study of southern species becomes more and more important. pathogen screenings in subspecies of apis mellifera capable of withstanding higher temperatures, provide an insight into future pathogen host interactions. screenings in different climate regions also provide a global perspective on the prevalence of certain pathogens. in this project, we performed a pathogen screening in apis mellif ...201627130035
effects of wintering environment and parasite-pathogen interactions on honey bee colony loss in north temperate regions.extreme winter losses of honey bee colonies are a major threat to beekeeping but the combinations of factors underlying colony loss remain debatable. we monitored colonies in two environments (colonies wintered indoors or outdoors) and characterized the effects of two parasitic mites, seven viruses, and nosema on honey bee colony mortality and population loss over winter. samples were collected from two locations within hives in fall, mid-winter and spring of 2009/2010. although fall parasite an ...201627448049
four categories of viral infection describe the health status of honey bee colonies.honey bee virus prevalence data are an essential prerequisite for managing epidemic events in a population. a survey study was carried out for seven viruses in colonies representing a healthy danish honey bee population. in addition, colonies from apiaries with high level varroa infestation or high level of winter mortality were also surveyed. results from rt-qpcr showed a considerable difference of virus levels between healthy and sick colonies. in the group of healthy colonies, no virus was de ...201526448627
honey bee apis mellifera parasites in the absence of nosema ceranae fungi and varroa destructor mites.few areas of the world have western honey bee (apis mellifera) colonies that are free of invasive parasites nosema ceranae (fungi) and varroa destructor (mites). particularly detrimental is v. destructor; in addition to feeding on host haemolymph, these mites are important vectors of several viruses that are further implicated as contributors to honey bee mortality around the world. thus, the biogeography and attendant consequences of viral communities in the absence of v. destructor are of sign ...201424955834
increased tolerance and resistance to virus infections: a possible factor in the survival of varroa destructor-resistant honey bees (apis mellifera).the honey bee ectoparasitic mite, varroa destructor, has a world-wide distribution and inflicts more damage than all other known apicultural diseases. however, varroa-induced colony mortality is more accurately a result of secondary virus infections vectored by the mite. this means that honey bee resistance to varroa may include resistance or tolerance to virus infections. the aim of this study was to see if this is the case for a unique population of mite-resistant (mr) european honey bees on t ...201424926792
cross-species transmission of honey bee viruses in associated arthropods.there are a number of rna virus pathogens that represent a serious threat to the health of managed honey bees (apis mellifera). that some of these viruses are also found in the broader pollinator community suggests the wider environmental spread of these viruses, with the potential for a broader impact on ecosystems. studies on the ecology and evolution of these viruses in the arthropod community as a whole may therefore provide important insights into these potential impacts. we examined manage ...201323845302
molecular identification of chronic bee paralysis virus infection in apis mellifera colonies in japan.chronic bee paralysis virus (cbpv) infection causes chronic paralysis and loss of workers in honey bee colonies around the world. although cbpv shows a worldwide distribution, it had not been molecularly detected in japan. our investigation of apis mellifera and apis cerana japonica colonies with rt-pcr has revealed cbpv infection in a. mellifera but not a. c. japonica colonies in japan. the prevalence of cbpv is low compared with that of other viruses: deformed wing virus (dwv), black queen cel ...201222852042
low prevalence of honeybee viruses in spain during 2006 and 2007.rna viruses that affect honeybees have been involved in colony losses reported around the world. the aim of the present work was to evaluate the prevalence and distribution of honeybee viruses during 2006-2007 in spanish professional apiaries, and their association with colony losses. four hundred and fifty-six samples from apiaries located in different geographic regions of spain were analyzed. thirty-seven percent of the samples had viral presence. most (80%) had one virus and 20% two differen ...201222513127
ictv virus taxonomy profile: iflaviridae.iflaviridae is a family of small non-enveloped viruses with monopartite, positive-stranded rna genomes of approximately 9-11 kilobases. viruses of all classified species infect arthropod hosts, with the majority infecting insects. both beneficial and pest insects serve as hosts, and infections can be symptomless (nilaparvatalugens honeydew virus 1) or cause developmental abnormalities (deformed wing virus), behavioural changes (sacbrood virus) and premature mortality (infectious flacherie virus) ...201728382900
the high-throughput production of dsrna against sacbrood virus for use in the honey bee apis cerana (hymenoptera: apidae).sacbrood virus (sbv) is a serious threat to honey bees. currently, there is no specific drug available for the treatment of sbv that does not affect the quality of the bee product. rna interference (rnai) is an important antiviral strategy for disease control. to effectively utilize this technology, the large-scale production and purification of double-stranded rna (dsrna) is necessary. here, a dsrna-expressing plasmid targeting the vp1 gene of chinese sacbrood virus (csbv) was constructed and e ...201627139728
correction: codon optimization, expression in escherichia coli, and immunogenicity of recombinant chinese sacbrood virus (csbv) structural proteins vp1, vp2, and vp3. 201526218595
codon optimization, expression in escherichia coli, and immunogenicity of recombinant chinese sacbrood virus (csbv) structural proteins vp1, vp2, and vp3.chinese sacbrood virus (csbv) is a small rna virus family belonging to the genus iflavirus that causes larval death, and even the collapse of entire bee colonies. the virus particle is spherical, non-enveloped, and its viral capsid is composed of four proteins, although the functions of the structural proteins are unclear. in this study, we used codon recoding to express the recombinant proteins vp1, vp2, and vp3 in escherichia coli. sds-page analysis and western blotting revealed that the targe ...201526067659
acute bee-paralysis virus in adult honey bees injected with sacbrood virus. 19676053149
one-step real-time quantitative pcr assays for the detection and field study of sacbrood honeybee and acute bee paralysis viruses.two one-step real-time rt-pcr assays, based on sybr green (sg) chemistry, were developed or adapted respectively, for the detection, differentiation, and quantitation of two important honeybee viruses: sacbrood virus (sbv) and acute bee paralysis virus (abpv). both reactions were optimized to yield the highest sensitivity and specificity. the genome equivalent copies (gec) detection limit per reaction was 389.3 for the abpv rt-pcr. the gec detection limit per reaction was 298.9 for the sbv rt-pc ...200919559729
honeybee viruses in uruguay.mortality of honeybees is a serious problem that beekeepers have to face periodically in uruguay and worldwide. the presence of rna viruses, in addition to other pathogens may be one of its possible causes. in this work, we detected chronic bee paralysis virus, acute bee paralysis virus, black queen cell virus, sacbrood virus and deformed wing virus in samples of uruguayan honeybees with or without varroa destructor and nosema apis. the detection of viruses in different provinces, simultaneous c ...200616843485
the nucleotide sequence of sacbrood virus of the honey bee: an insect picorna-like virus.we have determined the nucleotide sequence of sacbrood virus (sbv), which causes a fatal infection of honey bee larvae. the genomic rna of sbv is longer than that of typical mammalian picornaviruses (8832 nucleotides) and contains a single, large open reading frame (179-8752) encoding a polyprotein of 2858 amino acids. sequence comparison with other virus polyproteins revealed regions of similarity to characterized helicase, protease and rna-dependent rna polymerase domains; structural genes wer ...199910374974
complete genome of chinese sacbrood virus from apis cerana and analysis of the 3c-like cysteine protease.the complete genome sequence of chinese sacbrood virus (csbv), isolated from diseased larvae of apis cerana in fujian province, china was analyzed. the viral genome consisted of 8,800 nucleotides, encoding 2,848 amino acids. a phylogenetic tree analysis showed the sacbrood virus (sbv) segregated into three distinct groups. the isolates originated from a. c. indica and were the first distinct evolutionary group. the acsbv-sbm2 isolate, from a. c. cerana, belonged to the second distinct group. the ...201525557929
a novel strain of sacbrood virus of interest to world apiculture.this study has characterised a novel serotype of sacbrood virus (sbv) infecting apis mellifera in new guinea that has emerged in the presence of the introduced european and asian serotypes, which infect a. mellifera and apis cerana, respectively. the new guinea serotype appears to have evolved through mutation of the european serotype with no evidence of recombination between known strains, although recombination was detected in other sbv isolates from asia. sbv was also confirmed for the first ...201424650855
efficacy of silver ions against sacbrood virus infection in the eastern honey bee apis cerana.although silver is known to be a broad-spectrum biocidal agent, the effects of this metal against sacbrood virus have not yet been investigated. in this study, we evaluated the efficacy of silver ions against natural korean sacbrood virus (ksbv) infection of apis (a.) cerana. ten ksbv-infected colonies containing a. cerana with similar strength and activity were selected from an apiary located in bosung-gun (korea). among these, five colonies were randomly assigned to the treatment group that wa ...201525797295
characterization of gut bacteria at different developmental stages of asian honey bees, apis cerana.previous surveys have shown that adult workers of the asian honey bee apis cerana harbor four major gut microbes (bifidobacterium, snodgrassella alvi, gilliamella apicola, and lactobacillus). using quantitative pcr we characterized gut bacterial communities across the life cycle of a. cerana from larvae to workers. our results indicate that the presence and quantity of these four bacteria were low on day 1, increased rapidly after day 5, and then peaked during days 10-20. they stabilized from da ...201525805518
evidence of apis cerana sacbrood virus infection in apis mellifera.sacbrood virus(sbv) is one of the most destructive viruses in the asian honeybee apis cerana but is much less destructive in apis mellifera in previous studies, sbv isolates infecting a. cerana(acsbv) and sbv isolates infecting a. mellifera(amsbv) were identified as different serotypes, suggesting a species barrier in sbv infection. in order to investigate this species isolation, we examined the presence of sbv infection in 318a. mellifera colonies and 64a. cerana colonies, and we identified the ...201626801569
taqman mgb probe fluorescence real-time quantitative pcr for rapid detection of chinese sacbrood virus.sacbrood virus (sbv) is a picorna-like virus that affects honey bees (apis mellifera) and results in the death of the larvae. several procedures are available to detect chinese sbv (csbv) in clinical samples, but not to estimate the level of csbv infection. the aim of this study was develop an assay for rapid detection and quantification of this virus. primers and probes were designed that were specific for csbv structural protein genes. a taqman minor groove binder (mgb) probe-based, fluorescen ...201323408931
homology differences between complete sacbrood virus genomes from infected apis mellifera and apis cerana honeybees in korea.sacbrood virus (sbv) represents a serious threat to the health of managed honeybees. we determined four complete sbv genomic sequences (amsbv-kor1, amsbv-kor2, acsbv-kor3, and acsbv-kor4) isolated from apis mellifera and apis cerana in various regions of south korea. a phylogenetic tree was constructed from the complete genomic sequences of these korean sbvs (ksbvs) and 21 previously reported sbv sequences from other countries. three ksbvs (not amsbv-kor1) clustered with previously reported kore ...201626810400
in-situ hybridization for the detection of sacbrood virus in infected larvae of the honey bee (apis cerana).the aim of this study was to develop and use in-situ hybridization (ish) for the detection and localization of the sacbrood virus (sbv) in korean honey bee (apis cerana) larvae that were infected naturally with sbv. a 258 base pair cdna probe for sbv was generated by polymerase chain reaction. cells positive for viral genome typically showed a dark brown reaction in the cytoplasm. sbv was detected consistently in trophocytes and urocytes. the ish was successfully applied to routinely fixed and p ...201726852344
genomic characterization and phylogenetic analysis of chinese sacbrood virus isolated from loess plateau, china.the complete genomic rna of the chinese sacbrood virus (csbv) strain, which infects the honeybees in the loess plateau, was sequenced and analyzed. the csbv-sx strain contains 8705 nucleotides, which includes a single large open reading frame (99-8681 nucleotides) encoding 2860 amino acids. a novel efficient identification method was used to investigate the samples infected by csbv. the putative amino acid sequence alignment analysis showed that, except for some normal well characterized domains ...201627706762
a comparison of biological characteristics of three strains of chinese sacbrood virus in apis cerana.we selected and sequenced the entire genomes of three strains of chinese sacbrood virus (csbv): lnqy-2008 (isolated in qingyuan, liaoning province), sxyl-2015 (isolated in yulin, shanxi province), and jlcbs-2014 (isolated in changbaishan, jilin province), by vp1 amino acid (aa) analysis. these strains are endemic in china and infect apis cerana. nucleotide sequences, deduced amino acid sequences, genetic backgrounds, and other molecular biological characteristics were analysed. we also examined ...201627853294
phylogenetic analysis and survey of apis cerana strain of sacbrood virus (acsbv) in taiwan suggests a recent introduction.the sacbrood virus (sbv) is widely distributed in european honey bees, apis mellifera. acsbv, a distinct sbv strain in asian honey bees (a. cerana) causes larva death before pupation and often depopulates colonies, leading to collapse. it is the most severe disease in a. cerana beekeeping. acsbv infects a. cerana in most natural habitats, yet occurrences were not reported in taiwan before 2015 and were not a concern for local beekeepers. however, in 2016, a. cerana beekeepers in central taiwan r ...201728390783
inside honeybee hives: impact of natural propolis on the ectoparasitic mite varroa destructor and immunity is a key factor for honeybee health, including behavioral defense strategies such as the collective use of antimicrobial plant resins (propolis). while laboratory data repeatedly show significant propolis effects, field data are scarce, especially at the colony level. here, we investigated whether propolis, as naturally deposited in the nests, can protect honeybees against ectoparasitic mites varroa destructor and associated viruses, which are currently considered the most seriou ...201728178181
derivation of cell-adapted sacbrood virus (sbv) from the native korean honeybee.sacbrood virus (sbv), a causative agent of larval death in honeybees, is one of the most devastating diseases in bee industry throughout the world. lately the korean sacbrood virus (ksbv) induced great losses in korean honeybee (apis cerana) colonies. however, there is no culture system available for honeybee viruses, including sbv, therefore, the research on honeybee viruses is practically limited until present. in this study, we investigated the growth and replication of sbv in cell cultures. ...201525527463
new insights of sacbrood virus. 201425511927
rna viruses in hymenopteran pollinators: evidence of inter-taxa virus transmission via pollen and potential impact on non-apis hymenopteran species.although overall pollinator populations have declined over the last couple of decades, the honey bee (apis mellifera) malady, colony collapse disorder (ccd), has caused major concern in the agricultural community. among honey bee pathogens, rna viruses are emerging as a serious threat and are suspected as major contributors to ccd. recent detection of these viral species in bumble bees suggests a possible wider environmental spread of these viruses with potential broader impact. it is therefore ...201021203504
first molecular detection of a viral pathogen in ugandan honey bees.ugandan honey bees (apis mellifera l.) produce honey, and are key pollinators within commercial crops and natural ecosystems. real-time rt-pcr was used to screen immature and adult bees collected from 63 beekeeping sites across uganda for seven viral pathogens. no samples tested positive for chronic bee paralysis virus, sacbrood virus, deformed wing virus, acute bee paralysis virus, apis iridescent virus or israeli acute paralysis virus. however, black queen cell virus (bqcv) was found in 35.6% ...201020219470
occurrence and genetic analysis of picorna-like viruses infecting worker bees of apis mellifera l. populations in devon, south west england.viruses of the european honeybee, apis mellifera l. are known to reside at low levels in colonies, typically showing no apparent signs of infection. using reverse transcription-pcr (rt-pcr), 23 apiaries in devon were screened for the presence of 6 honeybee viruses, with positive colonies being analysed for viral genetic diversity. ninety-seven percent of the colonies were positive for deformed wing virus (dwv), 29% were positive for acute bee paralysis virus (abpv) and 1.4% were positive for bot ...200818359042
three-dimensional structure of the chinese sacbrood bee virus.the rna of chinese sacbrood bee virus (csbv) was purified and used as template to obtain a 1096 bp cdna fragment by rt-pcr amplification. this dna fragment was cloned into pgem-t easy vector for sequencing. analyses of the sequenced csbv rna fragment revealed a nucleotide sequence homology of 87.6% and a deduced amino-acid sequence homology of 94.6% with that of the sacbrood virus (sbv), indicating that csbv is a different but highly homologous virus of sbv. the three-dimensional (3d) structure ...200118726426
sacbrood virus of the honeybee (apis mellifera): rapid identification and phylogenetic analysis using reverse transcription-pcr.sacbrood virus (sbv) infects larvae of the honeybee (apis mellifera), resulting in failure to pupate and death. until now, identification of viruses in honeybee infections has been based on traditional methods such as electron microscopy, immunodiffusion, and enzyme-linked immunosorbent assay. culture cannot be used because no honeybee cell lines are available. these techniques are low in sensitivity and specificity. however, the complete nucleotide sequence of sbv has recently been determined, ...200111139201
the multiplication and spread of sacbrood virus of bees. 19695806364
the multiplication of sacbrood virus in the adult honeybee. 19685684126
viruslike particles in adult honey bees (apis mellifera linnaeus) following injection with sacbrood virus. 19676025061
sacbrood virus of the larval honey bee (apis mellifera linnaeus). 196414194138
the first comprehensive molecular detection of six honey bee viruses in iran in least 18 viruses have been reported in the honey bee (apis mellifera l.). however, severe diseases in honey bees are mainly caused by six viruses, and these are the most important in beekeeping. these viruses include: deformed wing virus (dwv), acute bee paralysis virus (abpv), chronic bee paralysis virus (cbpv), sacbrood virus (sbv), kashmir bee virus (kbv), and black queen cell virus (bqcv). in this study, we evaluated 89 iranian honey bee apiaries (during the period 2015-2016) suffering fr ...201728429131
viruses of managed alfalfa leafcutting bees (megachille rotundata fabricus) and honey bees (apis mellifera l.) in western canada: incidence, impacts, and prospects of cross-species viral transmission.we examined whether alfalfa leafcutting bees (alcb, megachille rotundata) experienced a higher incidence of seven viruses commonly found honey bees (apis mellifera) when placed alongside honey bees for hybrid canola seed pollination. although two viruses - sacbrood virus (sbv) and deformed wing virus (dwv) - were detected in alcb adults, their presence appeared independent of whether honey bees were present in the same field or not. a further survey of viruses among alcb adults in three differen ...201728400199
replication of chinese sacbrood virus in primary cell cultures of asian honeybee (apis cerana).a primary cell culture system was established for the first time from embryonic tissues of asian honeybee, apis cerana, and used to trace the early infection process of chinese sacbrood virus (csbv), an iflavirus in the family iflaviridae. a monolayer of epithelium-like cells of a. cerana, approximately 8-10 μm in diameter, was grown in kimura's insect medium at 28 °c within 3-4 days of setting up the cultures. such cultured cells were inoculated with csbv purified from infected larvae or pupae ...201425139546
proteomic analysis of apis cerana and apis mellifera larvae fed with heterospecific royal jelly and by csbv challenge.chinese honeybee apis cerana (ac) is one of the major asian honeybee species for local apiculture. however, ac is frequently damaged by chinese sacbrood virus (csbv), whereas apis mellifera (am) is usually resistant to it. heterospecific royal jelly (rj) breeding in two honeybee species may result in morphological and genetic modification. nevertheless, knowledge on the resistant mechanism of am to this deadly disease is still unknown. in the present study, heterospecific rj breeding was conduct ...201425102167
development and validation of a real-time two-step rt-qpcr taqman(®) assay for quantitation of sacbrood virus (sbv) and its application to a field survey of symptomatic honey bee colonies.sacbrood virus (sbv) is the causal agent of a disease of honey bee larvae, resulting in failure to pupate and causing death. the typical clinical symptom of sbv is an accumulation of sbv-rich fluid in swollen sub-cuticular pouches, forming the characteristic fluid-filled sac that gives its name to the disease. outbreaks of the disease have been reported in different countries, affecting the development of the brood and causing losses in honey bee colonies. today, few data are available on the sb ...201424121133
genetic characterization of vp1 gene of seven sacbrood virus isolated from three provinces in northern china during the years 2008-2012.the genetic diversity of seven northern china isolated sacbrood virus strains (sbv) has been analyzed, and hypervariable regions of the vp1 gene of 7 sbv were sequenced and characterized, in order to obtain epidemiological and immunological information, and to suggest typing criteria for sbv. sequence analysis of hypervariable regions of the vp1 gene in the genome of these isolates revealed a sequence homology of 91.0-99.3% among all seven local sbv isolates from apis cerana from china, with a s ...201323722004
an integrated proteomics reveals pathological mechanism of honeybee (apis cerena) sacbrood disease.viral diseases of honeybees are a major challenge for the global beekeeping industry. chinese indigenous honeybee (apis cerana cerana, acc) is one of the major asian honeybee species and has a dominant population with more than 3 million colonies. however, acc is frequently threatened by a viral disease caused by chinese sacbrood virus (csbv), which leads to fatal infections and eventually loss of the entire colony. nevertheless, knowledge on the pathological mechanism of this deadly disease is ...201323418745
complete genome sequence of sacbrood virus strain sbm2, isolated from the honeybee apis cerana in we report the complete genomic sequence of the sbm2 strain (vsbv-sbm2) of the sacbrood virus (sbv) that was isolated from the asian honeybee (apis cerana) in northern vietnam. the entire sequence excluding the 3' poly(a) tail is 8,834 nucleotides in length and contains a single large open reading frame (orf) of 8,580 nucleotides (position 178 to 8757), encoding 2,859 amino acids. vsbv-sbm2 shared 90 to 93% nucleotide identity and 95 to 96% amino acid homology to six complete genomes of sbv ...201323405307
first report of sacbrood virus in honey bee (apis mellifera) colonies in brazil.sacbrood disease, an affliction of honey bees (apis mellifera) characterized by brood that fails to pupate and subsequently dies, is an important threat to honey bee health. the disease is caused by the sacbrood virus (sbv), a positive-, single-stranded rna virus in the order picornavirales. because of the economic importance of honey bees for both pollination and honey production, it is vital to understand and monitor the spread of viruses such as sbv. this virus has been found in many places a ...201223079825
[new genotype of the sacbrood virus of the honeybee apis mellifera].the sacbrood virus (sbv) leads to death of honeybee larvae. until now, there were known three sbv genotypes: european, asian, and south african (e. grabensteiner et al., 2001; s. e. choe et al., 2012). serologic assay, sequencing and phylogenetic analysis of the sbv rna-polymerase gene fragments resulted in discovery of two genotypes of sbv circulating among the honeybee apis mellifera in the european region of russian federation (rf). one of them forms a new distinct genetic lineage of sbv (gen ...201222984772
reverse transcription loop-mediated isothermal amplification for sensitive and rapid detection of korean sacbrood virus.sacbrood virus (sbv) is one of the most serious honeybee viruses. the virus causes failure to pupate and death in both larvae and adult bees. recently, the korean sacbrood virus (ksbv) caused great losses in korean honeybee (apis cerana) colonies. although ksbv shows high homology with sbv strains, it has unique motifs and causes different symptoms. therefore, a simple, sensitive and specific method for detecting ksbv is needed urgently. in this study, a reverse transcription loop-mediated isoth ...201222947691
analysis of the complete genome sequence of two korean sacbrood viruses in the honey bee, apis mellifera.the complete genomic rnas of two korean sacbrood virus (sbv) strains, which infect the honey bee, apis mellifera, were sequenced. the two sequences (amsbv-kor19, amsbv-kor21) were distinguished by the presence or absence of a psti restriction site. these strains comprised of 8784 bp and 8835 bp; contained a single large orf (179-8707 and 179-8758) encoding 2843 and 2860 amino acids, respectively. deduced amino acid sequences comparison with some insect viruses showed that regions of helicase, pr ...201222749880
rapid detection of sacbrood virus (sbv) by one-step reverse transcription loop-mediated isothermal amplification assay.sacbrood virus (sbv) primarily infects honeybee broods, and in order to deal with the problem cost effective detection methods are required.201222340141
metatranscriptomic analyses of honey bee colonies.honey bees face numerous biotic threats from viruses to bacteria, fungi, protists, and mites. here we describe a thorough analysis of microbes harbored by worker honey bees collected from field colonies in geographically distinct regions of turkey. turkey is one of the world's most important centers of apiculture, harboring five subspecies of apis mellifera l., approximately 20% of the honey bee subspecies in the world. we use deep illumina-based rna sequencing to capture rna species for the hon ...201525852743
idiopathic brood disease syndrome and queen events as precursors of colony mortality in migratory beekeeping operations in the eastern united states.using standard epidemiological methods, this study set out to quantify the risk associated with exposure to easily diagnosed factors on colony mortality and morbidity in three migratory beekeeping operations. fifty-six percent of all colonies monitored during the 10-month period died. the relative risk (rr) that a colony would die over the short term (∼50 days) was appreciably increased in colonies diagnosed with idiopathic brood disease syndrome (ibds), a condition where brood of different ages ...201322939774
genetic and phylogenetic analysis of south korean sacbrood virus isolates from infected honey bees (apis cerana).sacbrood virus (sbv) is one of the most destructive honey bee viruses. the virus causes failure to pupate and death in both larvae and adult bees. genetic analysis of sbv infected honey bees (apis cerana) from five different provinces was carried out based on three nucleotide sequences; one partial structural protein coding sequence and two non-structural protein coding sequences. sequences amplified by three specific primer pairs were aligned and compared with reference sequences deposited in t ...201222221381
first molecular detection of co-infection of honey bee viruses in asymptomatic bombus atratus in south america.pollination is critical for food production and has the particularity of linking natural ecosystems with agricultural production systems. recently, losses of bumblebee species have been reported worldwide. in this study, samples from a commercial exploitation of bumblebees of argentina with a recent history of deaths were studied using a multiplex pcr for the detection of the honey bee viruses most frequently detected in south america. all samples analysed were positive for co-infections with de ...201324789396
honeybee colony disorder in crop areas: the role of pesticides and in many other locations in the world, honeybee colony losses and disorders have increased in belgium. some of the symptoms observed rest unspecific and their causes remain unknown. the present study aims to determine the role of both pesticide exposure and virus load on the appraisal of unexplained honeybee colony disorders in field conditions. from july 2011 to may 2012, 330 colonies were monitored. honeybees, wax, beebread and honey samples were collected. morbidity and mortality informatio ...201425048715
pathogen prevalence and abundance in honey bee colonies involved in almond pollination.honey bees are important pollinators of agricultural crops. since 2006, us beekeepers have experienced high annual honey bee colony losses, which may be attributed to multiple abiotic and biotic factors, including pathogens. however, the relative importance of these factors has not been fully elucidated. to identify the most prevalent pathogens and investigate the relationship between colony strength and health, we assessed pathogen occurrence, prevalence, and abundance in western us honey bee c ...201727053820
rapid detection of sacbrood virus in honeybee using ultra-rapid real-time polymerase chain reaction.a real-time reverse transcription-polymerase chain reaction (qrt-pcr) assay was developed for the fast and highly sensitive detection of the sacbrood virus (sbv) genome and applied to honeybee samples. using plasmid dna containing a partial sbv genome and diluted serially, as few as 1×10(2)copies/μl (correlation co-efficiency >0.99) were detected by the qrt-pcr assay, whereas 1×10(3)copies/μl were detected by the conventional rt-pcr assay. as a rapid detection method, ultra-rapid real-time pcr ( ...201222079620
the complete genome sequence of a single-stranded rna virus from the tarnished plant bug, lygus lineolaris (palisot de beauvois).the complete genome sequence of a single-stranded rna virus infecting the tarnished plant bug, lygus lineolaris (palisot de beauvois), was identified by sequencing cdna prepared from insects collected from the mississippi delta. the 9655 nucleotide positive-sense single-stranded rna genome of the l. lineolaris single-stranded rna virus (lylv-1) contained a single open reading frame of 8958 nucleotides encoding a 2986 amino acid genome polypeptide. the open reading frame was flanked by untranslat ...201221939663
acaricide treatment affects viral dynamics in varroa destructor-infested honey bee colonies via both host physiology and mite control.honey bee (apis mellifera) colonies are declining, and a number of stressors have been identified that affect, alone or in combination, the health of honey bees. the ectoparasitic mite varroa destructor, honey bee viruses that are often closely associated with the mite, and pesticides used to control the mite population form a complex system of stressors that may affect honey bee health in different ways. during an acaricide treatment using apistan (plastic strips coated with tau-fluvalinate), w ...201222020517
loop-mediated isothermal amplification for rapid detection of chinese sacbrood virus.chinese sacbrood virus (csbv) has emerged as an important etiologic agent of honeybee infections and is lethal for individual bees, even causing the collapse of entire colonies. although diagnostic methods for csbv have been established in many clinical laboratories, application of these methods is largely restricted by the apparatus needed to carry out the reaction and by cost, therefore a simpler and less expensive diagnostic method for csbv infection is required. in this study a simple and in ...201121640758
sudden deaths and colony population decline in greek honey bee colonies.during june and july of 2009, sudden deaths, tremulous movements and population declines of adult honey bees were reported by the beekeepers in the region of peloponnesus (mt. mainalo), greece. a preliminary study was carried out to investigate these unexplained phenomena in this region. in total, 37 bee samples, two brood frames containing honey bee brood of various ages, eight sugar samples and four sugar patties were collected from the affected colonies. the samples were tested for a range of ...201020804765
prevention of chinese sacbrood virus infection in apis cerana using rna interference.chinese sacbrood virus (csbv) is the pathogen of chinese sacbrood disease, which poses a serious threat to honeybee apis cerana, and tends to cause bee colony and even the whole apiary collapse. here we report on prevention of csbv infection by feeding second instar larvae of a. cerana with specific sequences of csbv double-stranded rna (dsrna). protection of the bee larvae from csbv by ingestion of csbv-derived dsrna was further demonstrated by quantitative real-time pcr (qrt-pcr) and northern ...201020379718
development of a multiplex rt-pcr for the simultaneous detection of three viruses of the honeybee (apis mellifera l.): acute bee paralysis virus, black queen cell virus and sacbrood virus.a single-step multiple-target (multiplex) reverse transcription-pcr (rt-pcr) was developed for the simultaneous detection and differentiation of three economically important viruses of the honeybee apis mellifera l.: acute bee paralysis virus (abpv), black queen cell virus (bqcv) and sacbrood virus (sbv). three compatible sets of primers, specific for each virus, were designed in conserved regions of the viral genomes for use in a one-step (single tube) rt-pcr assay. the individual rt-pcr assays ...200717207498
absence of deformed wing virus and varroa destructor in australia provides unique perspectives on honeybee viral landscapes and colony losses.honeybee (apis mellifera) health is threatened globally by the complex interaction of multiple stressors, including the parasitic mite varroa destructor and a number of pathogenic viruses. australia provides a unique opportunity to study this pathogenic viral landscape in the absence of v. destructor. we analysed 1,240a. mellifera colonies across australia by reverse transcription-polymerase chain reaction (rt-pcr) and next-generation sequencing (ngs). five viruses were prevalent: black queen ce ...201728761114
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