Communications in agricultural and applied biological sciences
A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful... more A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful either as template for RT-PCR or in nucleic acids hybridization, was developed. Sensitivity of the methods were evaluated up to 10(9) and 10(6) dilution of TNAs extracted from a single honeybee, for reverse transcriptase polymerase chain reaction and molecular hybridization respectively. The two diagnostic methods developed could be useful for the study of the molecular biology and the pathology of DWV. For practical applications dot-blot hybridization could be used in order to study the incidence of DWV in honeybees populations. The method is enough sensitive, rapid and less affected by contamination problems compared to RT-PCR and thus it could be applied to the sanitary certification of honeybees and their products.
The importance of honeybee products make of disease prevention and control in honeybees one of th... more The importance of honeybee products make of disease prevention and control in honeybees one of the mainconcerns of beekeepers in the world. The PCR – RT reaction represents an alternative for amplification performed inorder to realize the Varroa destructor O. genotypization, very important stage in haoneybee resistance to parasitedescription and also in management of the treatments. The linearity data is a very important parameter and very usefulin determination of the amplification of the parasite DNA and success of the genotypization process. The amplificationefficiency was very satisfactory, fact revealed by the value of the regression line y = - 2.3103 * 26.552 together withcoefficient of determination equal (r2 = 0.9691), meaning that more than 96% of the reaction efficiency may beexplained by the process liniarity. The implementation of the RT-PCR method was successful and it represents apremise for validation process evolution.
Mites in the genus Varroa are obligate ectoparasites of honey bee populations worldwide. Recent e... more Mites in the genus Varroa are obligate ectoparasites of honey bee populations worldwide. Recent evidence from morphological, geographical, and especially genetic variation has spurred an important revision of Varroa taxonomy. Specifically, mitochondrial DNA (mtDNA) evidence suggests that the main mite pest on western honey bees (Apis mellifera) is not Varroa jacobsoni, as first described, but a distinct species now named Varroa destructor. Genetic markers also have been used to support a taxonomic basis for regional differences in how Varroa mites impact honey bees. Recent morphometric and molecular studies confirmed the presence of the species V. destructor also in the apiaries of the Campania region of southern Italy. In the three-year period 2001-2003 a survey was conducted in 118 municipalities of the five provinces of the Campania region in order to add data to the limited epidemiological information available regarding Varroa destructor in this zone. The level of infestation b...
Communications in agricultural and applied biological sciences, 2006
A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful... more A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful either as template for RT-PCR or in nucleic acids hybridization, was developed. Sensitivity of the methods were evaluated up to 10(9) and 10(6) dilution of TNAs extracted from a single honeybee, for reverse transcriptase polymerase chain reaction and molecular hybridization respectively. The two diagnostic methods developed could be useful for the study of the molecular biology and the pathology of DWV. For practical applications dot-blot hybridization could be used in order to study the incidence of DWV in honeybees populations. The method is enough sensitive, rapid and less affected by contamination problems compared to RT-PCR and thus it could be applied to the sanitary certification of honeybees and their products.
The health of the honeybee and, indirectly, global crop production are threatened by several biot... more The health of the honeybee and, indirectly, global crop production are threatened by several biotic and abiotic factors, which play a poorly defined role in the induction of widespread colony losses. Recent descriptive studies suggest that colony losses are often related to the interaction between pathogens and other stress factors, including parasites. Through an integrated analysis of the population and molecular changes associated with the collapse of honeybee colonies infested by the parasitic mite Varroa destructor, we show that this parasite can de-stabilise the within-host dynamics of Deformed wing virus (DWV), transforming a cryptic and vertically transmitted virus into a rapidly replicating killer, which attains lethal levels late in the season. The de-stabilisation of DWV infection is associated with an immunosuppression syndrome, characterized by a strong down-regulation of the transcription factor NF-κB. The centrality of NF-κB in host responses to a range of environment...
The importance of honeybee products make of disease prevention and control in honeybees one of th... more The importance of honeybee products make of disease prevention and control in honeybees one of the mainconcerns of beekeepers in the world. The PCR RT reaction represents an alternative for amplification performed inorder to realize the Varroa destructor O. genotypization, very ...
Honey bees (Apis mellifera L.) are bioindicators of environmental pollution levels. During their
... more Honey bees (Apis mellifera L.) are bioindicators of environmental pollution levels. During their wide-ranging foraging activity, these hymenopterans are exposed to pollutants, thus becoming a useful tool to trace the environmental contaminants as heavy metals, pesticides, radionuclides and volatile organic compounds. In the present work we demonstrate that bees can also be used as active samplers of airborne particulate matter. Worker bees were collected from hives located in a polluted postmining area in South West Sardinia (Italy) that is also exposed to dust emissions from industrial plants. The area is included in an official list of sites of national interest for environmental remediation, and has been characterized for the effects of pollutants on the health of the resident population. The head, wings, hind legs and alimentary canal of the bees were investigated with Scanning ElectronMicroscopy coupled with Xray spectroscopy (SEM-EDX). The analyses pointed to specific morphological and chemical features of the particulate, and resulted into the identification of three categories of particles: industry -, postmining -, and soil –derived. With the exception of the gut, all the analyzed body districts displayed inorganic particles, mostly concentrated in specific areas of the body (i.e. along the costal margin of the fore wings, the medial plane of the head, and the inner surface of the hind legs). The role of both past mining activities and the industrial activity close to the study area as sources of the particulate matter is also discussed. We conclude that honey bees are able to collect samples of the main airborne particles emitted from different sources, therefore could be an ideal tool for monitoring such a kind of pollutants.
The dynamics of viruses are critical to our understanding of disease pathogenesis. Using honey be... more The dynamics of viruses are critical to our understanding of disease pathogenesis. Using honey bee Deformed wing virus (DWV) as a model, we conducted field and laboratory studies to investigate the roles of abiotic and biotic stress factors as well as host health conditions in dynamics of virus replication in honey bees. The results showed that temperature decline could lead to not only significant decrease in the rate for pupae to emerge as adult bees, but also an increased severity of the virus infection in emerged bees, partly explaining the high levels of winter losses of managed honey bees, Apis mellifera, around the world. By experimentally exposing adult bees with variable levels of parasitic mite Varroa destructor, we showed that the severity of DWV infection was positively correlated with the density and time period of Varroa mite infestation, confirming the role of Varroa mites in virus transmission and activation in honey bees. Further, we showed that host conditions have...
Communications in agricultural and applied biological sciences
A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful... more A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful either as template for RT-PCR or in nucleic acids hybridization, was developed. Sensitivity of the methods were evaluated up to 10(9) and 10(6) dilution of TNAs extracted from a single honeybee, for reverse transcriptase polymerase chain reaction and molecular hybridization respectively. The two diagnostic methods developed could be useful for the study of the molecular biology and the pathology of DWV. For practical applications dot-blot hybridization could be used in order to study the incidence of DWV in honeybees populations. The method is enough sensitive, rapid and less affected by contamination problems compared to RT-PCR and thus it could be applied to the sanitary certification of honeybees and their products.
The importance of honeybee products make of disease prevention and control in honeybees one of th... more The importance of honeybee products make of disease prevention and control in honeybees one of the mainconcerns of beekeepers in the world. The PCR – RT reaction represents an alternative for amplification performed inorder to realize the Varroa destructor O. genotypization, very important stage in haoneybee resistance to parasitedescription and also in management of the treatments. The linearity data is a very important parameter and very usefulin determination of the amplification of the parasite DNA and success of the genotypization process. The amplificationefficiency was very satisfactory, fact revealed by the value of the regression line y = - 2.3103 * 26.552 together withcoefficient of determination equal (r2 = 0.9691), meaning that more than 96% of the reaction efficiency may beexplained by the process liniarity. The implementation of the RT-PCR method was successful and it represents apremise for validation process evolution.
Mites in the genus Varroa are obligate ectoparasites of honey bee populations worldwide. Recent e... more Mites in the genus Varroa are obligate ectoparasites of honey bee populations worldwide. Recent evidence from morphological, geographical, and especially genetic variation has spurred an important revision of Varroa taxonomy. Specifically, mitochondrial DNA (mtDNA) evidence suggests that the main mite pest on western honey bees (Apis mellifera) is not Varroa jacobsoni, as first described, but a distinct species now named Varroa destructor. Genetic markers also have been used to support a taxonomic basis for regional differences in how Varroa mites impact honey bees. Recent morphometric and molecular studies confirmed the presence of the species V. destructor also in the apiaries of the Campania region of southern Italy. In the three-year period 2001-2003 a survey was conducted in 118 municipalities of the five provinces of the Campania region in order to add data to the limited epidemiological information available regarding Varroa destructor in this zone. The level of infestation b...
Communications in agricultural and applied biological sciences, 2006
A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful... more A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful either as template for RT-PCR or in nucleic acids hybridization, was developed. Sensitivity of the methods were evaluated up to 10(9) and 10(6) dilution of TNAs extracted from a single honeybee, for reverse transcriptase polymerase chain reaction and molecular hybridization respectively. The two diagnostic methods developed could be useful for the study of the molecular biology and the pathology of DWV. For practical applications dot-blot hybridization could be used in order to study the incidence of DWV in honeybees populations. The method is enough sensitive, rapid and less affected by contamination problems compared to RT-PCR and thus it could be applied to the sanitary certification of honeybees and their products.
The health of the honeybee and, indirectly, global crop production are threatened by several biot... more The health of the honeybee and, indirectly, global crop production are threatened by several biotic and abiotic factors, which play a poorly defined role in the induction of widespread colony losses. Recent descriptive studies suggest that colony losses are often related to the interaction between pathogens and other stress factors, including parasites. Through an integrated analysis of the population and molecular changes associated with the collapse of honeybee colonies infested by the parasitic mite Varroa destructor, we show that this parasite can de-stabilise the within-host dynamics of Deformed wing virus (DWV), transforming a cryptic and vertically transmitted virus into a rapidly replicating killer, which attains lethal levels late in the season. The de-stabilisation of DWV infection is associated with an immunosuppression syndrome, characterized by a strong down-regulation of the transcription factor NF-κB. The centrality of NF-κB in host responses to a range of environment...
The importance of honeybee products make of disease prevention and control in honeybees one of th... more The importance of honeybee products make of disease prevention and control in honeybees one of the mainconcerns of beekeepers in the world. The PCR RT reaction represents an alternative for amplification performed inorder to realize the Varroa destructor O. genotypization, very ...
Honey bees (Apis mellifera L.) are bioindicators of environmental pollution levels. During their
... more Honey bees (Apis mellifera L.) are bioindicators of environmental pollution levels. During their wide-ranging foraging activity, these hymenopterans are exposed to pollutants, thus becoming a useful tool to trace the environmental contaminants as heavy metals, pesticides, radionuclides and volatile organic compounds. In the present work we demonstrate that bees can also be used as active samplers of airborne particulate matter. Worker bees were collected from hives located in a polluted postmining area in South West Sardinia (Italy) that is also exposed to dust emissions from industrial plants. The area is included in an official list of sites of national interest for environmental remediation, and has been characterized for the effects of pollutants on the health of the resident population. The head, wings, hind legs and alimentary canal of the bees were investigated with Scanning ElectronMicroscopy coupled with Xray spectroscopy (SEM-EDX). The analyses pointed to specific morphological and chemical features of the particulate, and resulted into the identification of three categories of particles: industry -, postmining -, and soil –derived. With the exception of the gut, all the analyzed body districts displayed inorganic particles, mostly concentrated in specific areas of the body (i.e. along the costal margin of the fore wings, the medial plane of the head, and the inner surface of the hind legs). The role of both past mining activities and the industrial activity close to the study area as sources of the particulate matter is also discussed. We conclude that honey bees are able to collect samples of the main airborne particles emitted from different sources, therefore could be an ideal tool for monitoring such a kind of pollutants.
The dynamics of viruses are critical to our understanding of disease pathogenesis. Using honey be... more The dynamics of viruses are critical to our understanding of disease pathogenesis. Using honey bee Deformed wing virus (DWV) as a model, we conducted field and laboratory studies to investigate the roles of abiotic and biotic stress factors as well as host health conditions in dynamics of virus replication in honey bees. The results showed that temperature decline could lead to not only significant decrease in the rate for pupae to emerge as adult bees, but also an increased severity of the virus infection in emerged bees, partly explaining the high levels of winter losses of managed honey bees, Apis mellifera, around the world. By experimentally exposing adult bees with variable levels of parasitic mite Varroa destructor, we showed that the severity of DWV infection was positively correlated with the density and time period of Varroa mite infestation, confirming the role of Varroa mites in virus transmission and activation in honey bees. Further, we showed that host conditions have...
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wide-ranging foraging activity, these hymenopterans are exposed to pollutants, thus becoming
a useful tool to trace the environmental contaminants as heavy metals, pesticides, radionuclides
and volatile organic compounds. In the present work we demonstrate that bees can
also be used as active samplers of airborne particulate matter. Worker bees were collected
from hives located in a polluted postmining area in South West Sardinia (Italy) that is also
exposed to dust emissions from industrial plants. The area is included in an official list of sites
of national interest for environmental remediation, and has been characterized for the effects
of pollutants on the health of the resident population. The head, wings, hind legs and alimentary
canal of the bees were investigated with Scanning ElectronMicroscopy coupled with Xray
spectroscopy (SEM-EDX). The analyses pointed to specific morphological and chemical
features of the particulate, and resulted into the identification of three categories of particles:
industry -, postmining -, and soil –derived. With the exception of the gut, all the analyzed body
districts displayed inorganic particles, mostly concentrated in specific areas of the body (i.e.
along the costal margin of the fore wings, the medial plane of the head, and the inner surface
of the hind legs). The role of both past mining activities and the industrial activity close to the
study area as sources of the particulate matter is also discussed. We conclude that honey
bees are able to collect samples of the main airborne particles emitted from different sources,
therefore could be an ideal tool for monitoring such a kind of pollutants.
wide-ranging foraging activity, these hymenopterans are exposed to pollutants, thus becoming
a useful tool to trace the environmental contaminants as heavy metals, pesticides, radionuclides
and volatile organic compounds. In the present work we demonstrate that bees can
also be used as active samplers of airborne particulate matter. Worker bees were collected
from hives located in a polluted postmining area in South West Sardinia (Italy) that is also
exposed to dust emissions from industrial plants. The area is included in an official list of sites
of national interest for environmental remediation, and has been characterized for the effects
of pollutants on the health of the resident population. The head, wings, hind legs and alimentary
canal of the bees were investigated with Scanning ElectronMicroscopy coupled with Xray
spectroscopy (SEM-EDX). The analyses pointed to specific morphological and chemical
features of the particulate, and resulted into the identification of three categories of particles:
industry -, postmining -, and soil –derived. With the exception of the gut, all the analyzed body
districts displayed inorganic particles, mostly concentrated in specific areas of the body (i.e.
along the costal margin of the fore wings, the medial plane of the head, and the inner surface
of the hind legs). The role of both past mining activities and the industrial activity close to the
study area as sources of the particulate matter is also discussed. We conclude that honey
bees are able to collect samples of the main airborne particles emitted from different sources,
therefore could be an ideal tool for monitoring such a kind of pollutants.