Complete genome sequence of deformed wing virus and black queen cell virus isolated from honeybees (Apis mellifera) in Argentina.

We report the complete genome sequence of deformed wing virus and black queen cell virus isolated from Argentinean's honeybees. These sequence data will be valuable for future research on the viral variants present in the country and the development of strategies to control the spread of these viruses in apiaries.

Prevalence of Adult Honey Bee (Apis mellifera L.) Pests and Pathogens in the Five Beekeeping Regions of Mexico.

Mexico is a major honey producer, but not much information exists about the health status of honey bees (Apis mellifera L.) in the country. This study was conducted to determine the sanitary status of adult honey bees in Mexico's five beekeeping regions. Samples from 369 apiaries were diagnosed to identify pathogens such as Varroa destructor, which was quantified, Acarapis woodi, Nosema spp., and five viruses. Colonies were also inspected for the presence of the small hive beetle (SHB), Aethina tumida. Varroa destructor was found in 83.5% of the apiaries, with the Pacific Coast region having the highest prevalence (>95%) and rates (4.5% ± 0.6). Acarapis woodi was detected in only one apiary from the Pacific Coast, whereas Nosema spp. were prevalent in 48.5% of the apiaries, with the highest and lowest frequencies in the Yucatan Peninsula and North regions (64.6% and 10.2%, respectively). For viruses, deformed wing virus (DWV) was detected in 26.1% of the apiaries, with the highest frequency in the Pacific Coast region (44.7%). Israeli acute paralysis virus (IAPV) was diagnosed in 3.2% of the samples and sacbrood bee virus (SBV) in 23.3% of them, with the highest frequency in the High Plateau region (36.4%). Chronic bee paralysis and Kashmir bee viruses were not detected. SHB prevalence was 25.2% nationwide, with the highest frequency in the Yucatan Peninsula (39.2%). This study shows that the most common parasites of adult honey bees in Mexico are V. destructor and Nosema spp., and that the most prevalent virus is DWV, whereas SHB is highly prevalent in the Yucatan Peninsula. This information could be useful to design disease control strategies for honey bee colonies in different regions of Mexico.

Effect of Chronic Exposure to Sublethal Doses of Imidacloprid and Nosema ceranae on Immunity, Gut Microbiota, and Survival of Africanized Honey Bees.

Large-scale honey bee colony losses reported around the world have been associated with intoxication with pesticides, as with the presence of pests and pathogens. Among pesticides, neonicotinoid insecticides are the biggest threat. Due to their extensive use, they can be found in all agricultural environments, including soil, water, and air, are persistent in the environment, and are highly toxic for honey bees. In addition, infection by different pests and pathogens can act synergistically, weakening bees. In this study, we investigated the effects of chronic exposure to sublethal doses of imidacloprid alone or combined with the microsporidia Nosema ceranae on the immune response, deformed wing virus infection (DWV), gut microbiota, and survival of Africanized honey bees. We found that imidacloprid affected the expression of some genes associated with immunity generating an altered physiological state, although it did not favor DWV or N. ceranae infection. The pesticide alone did not affect honey bee gut microbiota, as previously suggested, but when administered to N. ceranae infected bees, it generated significant changes. Finally, both stress factors caused high mortality rates. Those results illustrate the negative impact of imidacloprid alone or combined with N. ceranae on Africanized honey bees and are useful to understand colony losses in Latin America.

Warmer winters are associated with lower levels of the cryoprotectant glycerol, a slower decrease in vitellogenin expression and reduced virus infections in winter honeybees.

Within the context of climate change, winter temperatures at high latitudes are predicted to rise faster than summer temperatures. This phenomenon is expected to negatively affect the diapause performance and survival of insects, since they largely rely on low temperatures to lower their metabolism and preserve energy. However, some insects like honeybees, remain relatively active during the winter and elevate their metabolic rate to produce endothermic heat when temperatures drop. Warming winters are thus expected to improve overwintering performance of honeybees. In order to verify this hypothesis, for two consecutive years, we exposed honeybee colonies to either a mild or cold winter. We then monitored the influence of wintering conditions on several parameters of honeybee overwintering physiology, such as levels of the cryoprotectant glycerol, expression levels of immune and antioxidant genes, and genes encoding multifunctional proteins, including vitellogenin, which promotes bee longevity. Winter conditions had no effect on the expression of antioxidant genes, and genes related to immunity were not consistently affected. However, mild winters were consistently associated with a lower investment in glycerol synthesis and a higher expression of fat body genes, especially apidaecin and vitellogenin. Finally, while we found that viral loads generally decreased through the winter, this trend was more pronounced under mild winter conditions. In conclusion, and without considering how warming temperatures might affect other aspects of honeybee biology before overwintering, our data suggest that warming temperatures will likely benefit honeybee vitality by notably reducing their viral loads over the winter.

Impact of Chronic Exposure to Sublethal Doses of Glyphosate on Honey Bee Immunity, Gut Microbiota and Infection by Pathogens.

Glyphosate is the most used pesticide around the world. Although different studies have evidenced its negative effect on honey bees, including detrimental impacts on behavior, cognitive, sensory and developmental abilities, its use continues to grow. Recent studies have shown that it also alters the composition of the honey bee gut microbiota. In this study we explored the impact of chronic exposure to sublethal doses of glyphosate on the honey bee gut microbiota and its effects on the immune response, infection by Nosema ceranae and Deformed wing virus (DWV) and honey bee survival. Glyphosate combined with N. ceranae infection altered the structure and composition of the honey bee gut microbiota, for example by decreasing the relative abundance of the core members Snodgrassella alvi and Lactobacillus apis. Glyphosate increased the expression of some immune genes, possibly representing a physiological response to mitigate its negative effects. However, this response was not sufficient to maintain honey bee health, as glyphosate promoted the replication of DWV and decreased the expression of vitellogenin, which were accompanied by a reduced life span. Infection by N. ceranae also alters honey bee immunity although no synergistic effect with glyphosate was observed. These results corroborate previous findings suggesting deleterious effects of widespread use of glyphosate on honey bee health, and they contribute to elucidate the physiological mechanisms underlying a global decline of pollination services.

Deformed wing virus type a and b in managed honeybee colonies of Argentina.

Apis mellifera is infected by more than 24 virus species worldwide, mainly positive-sense, single-stranded RNA viruses of the Dicistroviridae and Iflaviridae families. Among the viruses that infect honey bees, Deformed wing virus is the most prevalent and is present as three master variants DWV-A, B, and C. Given that the ectoparasitic mite Varroa destructor vectors these virus variants, recombination events between them are expected, and variants and their recombinants can co-exist in mites and honeybees at the same time. In this study, we detect, through RT-qPCR, the presence of DWV-A and B in the same samples of adult bees from colonies of Argentina. Total RNA was extracted from pools of ten adult bees from 45 apiaries distributed across the main beekeeping Provinces of Argentina (Buenos Aires, Santa Fe, Córdoba, Santiago del Estero, Río Negro, and Mendoza); then RT-qPCR reactions were performed to detect DWV-A and B, with specific primer pairs. After the amplifications, PCR products (204 and 660 bp amplicons for DWV-B, and ~250 bp for DWV-A) were purified and sequenced to verify that they corresponded to reported sequences, analyzing them using the Blast software. Of the 45 samples analyzed by RT-qPCR, over 90% were infected with DWV-A and 47% were also positive for DWV-B, where it was found in high prevalence specifically in colonies of A. mellifera of the Buenos Aires Province. Future studies will determine the impact of this type of the virus and its ability to recombine with the other DWV types in the apiaries of our country.

Evidence of presence and replication of honey bee viruses among wild bee pollinators in subtropical environments.

We determined the presence of six viruses in different bee species collected in subtropical environments. Deformed wing virus (DWV) and black queen cell virus (BQCV) were detected in >90% of honey bee samples and in 50-100% of four stingless bee, two bumble bee and one solitary bee species. Additionally, minus DWV and BQCV RNA strands were detected, indicating that the viruses replicate in several hosts. This is the first report of honey bee viruses replicating in six wild bee species in the tropics. If pathogenic to them, viral infections could result in negative impacts in agricultural and unmanaged ecosystems.

Impact of Varroa destructor and deformed wing virus on emergence, cellular immunity, wing integrity and survivorship of Africanized honey bees in Mexico.

The ectoparasitic mite Varroa destructor is the primary health problem of honey bees (Apis mellifera) worldwide. Africanized honey bees in Brazil have demonstrated tolerance to the mite, but there is controversy about the degree of mite tolerance of Africanized bees in other countries. This study was conducted to quantify the effect of V. destructor parasitism on emergence, hemocyte concentration, wing integrity and longevity of Africanized honey bees in Mexico. Africanized bee brood were artificially infested with V. destructor mites and held in an incubator until emergence as adults and compared to non-infested controls. Deformed wing virus (DWV) presence was determined in the mites used to infest the bees. After emergence, the bees were maintained in an incubator to determine survivorship. The percentage of worker bees that emerged from parasitized cells (69%) was significantly lower than that of bees emerged from non-infested cells (96%). Newly-emerged parasitized bees had a significantly lower concentration of hemocytes in the hemolymph than non-parasitized bees. Additionally, the proportion of bees with deformed wings that emerged from V. destructor-parasitized cells was significantly higher (54%) than that of the control group (0%). The mean survival time of bees that emerged from infested and non-infested cells was 8.5 ±â€¯0.3 and 14.4 ±â€¯0.4 days, respectively, and the difference was significant. We conclude that V. destructor parasitism and DWV infections kill, cause deformities and inhibit cellular immunity in developing Africanized honey bees, and significantly reduce the lifespan of adult bees in Mexico. These results suggest that the tolerance of Africanized bees to V. destructor is related to adult bee mechanisms.

Prevalence of honey bee (Apis mellifera) viruses in temperate and subtropical regions from Argentina / Prevalencia de virus en abejas melíferas (Apis mellifera) en ambientes templados y subtropicales de Argentina

In Argentina, bee virus studies are still incipient, and there are no studies regarding the climatic effect. The aim of this study was to assess and compare the presence of honeybee viruses in different climatic regions from Argentina. A total of 385 colonies distributed in five Argentinean eco-regions were examined to evaluate the percentage of infestation with Varroa destructor and the presence of seven virus species (Deformed wing virus, DWV; Acute bee paralysis virus, ABPV; Chronic bee paralysis virus, CBPV; Black queen cell virus, BQCV; Kashmer bee virus, KBV; Israeli acute bee paralysis virus, IAPV; and Sacbrood bee virus, SBV) after honey yield. Two viruses, KBV and IAPV, were not detected. The other five viruses were found in different prevalences: DWV (35%), ABPV (21.5%), BQCV (8.0%), CBPV (2.2%), and SBV (1.1%). We found double and triple viral associations in approximately 25% of the sampled colonies. The mean V. destructor infestation in the colonies prior to the acaricide treatment was 7.12% ± 8.7%. The knowledge of the prevalence of these viruses in the region and their relation with the mite and other possible influencing factors is important for preventing colony losses. Further studies are necessary to identify the risk factors associated with virus presence and its relationship with other pathogens such as V. destructor.

En Argentina, los estudios sobre prevalencia de virus en abejas continúan siendo incipientes y no existen reportes acerca de cómo inciden sobre dicha prevalencia las variables climáticas. El objetivo de este estudio fue evaluar y comparar la presencia de virus en abejas melíferas en diferentes regiones agroecológicas de Argentina. A tal fin se evaluaron 385 colmenas distribuidas en 5 regiones agroecológicas de las provincias de Chaco y Santa Fe; en ellas se analizó el porcentaje de infestación con Varroa destructor (ácaro patógeno de abejas) y la presencia de 7 especies de virus (DWV, virus de las alas deformadas; ABPV, virus de la parálisis aguda de la abeja; CBPV, virus de la parálisis crónica de la abeja; BQCV, virus de celda negra de la reina; KBV, virus de la abeja de Cachemira; IAPV, virus israelí de la parálisis aguda y SBV, virus de la cría ensacada). luego de la cosecha de miel. Dos virus (KBV y IAPV) no fueron detectados. Las otras 5 especies de virus se encontraron con prevalencias variables: DWV (35%), ABPV (21,5%), BQCV (8%), CBPV (2,2%) y SBV (1,1%). Fue posible identificar la presencia de 3 y hasta 3 virus simultáneamente en el 25% de las colmenas evaluadas. El promedio de infestación por V. destructor en las colmenas luego de la cosecha de miel y antes del tratamiento con acaricidas fue de 7,12% (±8,7). Conocer la prevalencia de virus en las diferentes regiones agroecológicas y su relación con la presencia del ácaro V. destructor e identificar otros posibles factores que podrían influir en su presencia es relevante para definir estrategias que reduzcan la mortandad de colmenas. Es necesario realizar estudios adicionales para identificar los factores de riesgo asociados a la presencia de virus en las colmenas y su relación con otros patógenos, como V. destructor.

Risk factors for the presence of Deformed wing virus and Acute bee paralysis virus under temperate and subtropical climate in Argentinian bee colonies.

Beekeepers all across the world are suffering important losses of their colonies, and the parasitic mites Varroa destructor and Nosema sp, as well as several bee viruses, are being pointed out as the possible causes of these losses, generally associated with environmental and management factors. The objective of the present study was to evaluate the presence of seven virus species (Deformed wing virus -DWV-, Acute bee paralysis virus -ABPV-, Chronic bee paralysis virus -CBPV-, Black queen cell virus -BQCV-, Kashmir bee virus -KBV-, Israeli acute bee paralysis virus -IAPV-, and Sacbrood bee virus -SBV), as well as the prevalence of Nosema sp. and Varroa destructor, and their possible associated factors, under temperate and subtropical climate conditions in Argentinean colonies. A total of 385 colonies distributed in five Argentinean eco-regions were examined after honey harvest. The final multivariable model revealed only one variable associated with the presence of DWV and two with the presence of ABPV. The apiary random effect was significant in both cases (P=0.018; P=0.006, respectively). Colonies with a Varroa infestation rate >3% showed higher presence of DWV than colonies with <3% of Varroa infestation level (OR=1.91; 95% CI: 1.02-3.57; P<0.044). The same pattern was observed for the presence of ABPV (OR=2.23; 95% CI: 1.04-4.77; P<0.039). Also, colonies where replacement of old combs was not a common practice had higher presence of ABPV (OR=6.02; 95% CI: 1.16-31.25; P<0.033). Regardless of the location of the colonies, virus presence was strongly associated with V. destructor level. Therefore, all the factors that directly or indirectly influence the levels of mites will be also influencing the presence of the viruses.

Prevalence of honey bee (Apis mellifera) viruses in temperate and subtropical regions from Argentina.

In Argentina, bee virus studies are still incipient, and there are no studies regarding the climatic effect. The aim of this study was to assess and compare the presence of honeybee viruses in different climatic regions from Argentina. A total of 385 colonies distributed in five Argentinean eco-regions were examined to evaluate the percentage of infestation with Varroa destructor and the presence of seven virus species (Deformed wing virus, DWV; Acute bee paralysis virus, ABPV; Chronic bee paralysis virus, CBPV; Black queen cell virus, BQCV; Kashmer bee virus, KBV; Israeli acute bee paralysis virus, IAPV; and Sacbrood bee virus, SBV) after honey yield. Two viruses, KBV and IAPV, were not detected. The other five viruses were found in different prevalences: DWV (35%), ABPV (21.5%), BQCV (8.0%), CBPV (2.2%), and SBV (1.1%). We found double and triple viral associations in approximately 25% of the sampled colonies. The mean V. destructor infestation in the colonies prior to the acaricide treatment was 7.12%±8.7%. The knowledge of the prevalence of these viruses in the region and their relation with the mite and other possible influencing factors is important for preventing colony losses. Further studies are necessary to identify the risk factors associated with virus presence and its relationship with other pathogens such as V. destructor.

Varroa destructor (Mesostigmata: Varroidae) Parasitism and Climate Differentially Influence the Prevalence, Levels, and Overt Infections of Deformed Wing Virus in Honey Bees (Hymenoptera: Apidae).

The prevalence and loads of deformed wing virus (DWV) between honey bee (Apis mellifera L.) colonies from a tropical and a temperate environment were compared. The interaction between these environments and the mite Varroa destructor in relation to DWV prevalence, levels, and overt infections, was also analyzed. V. destructor rates were determined, and samples of mites, adult bees, brood parasitized with varroa mites and brood not infested by mites were analyzed. DWV was detected in 100% of the mites and its prevalence and loads in honey bees were significantly higher in colonies from the temperate climate than in colonies from the tropical climate. Significant interactions were found between climate and type of sample, with the highest levels of DWV found in varroa-parasitized brood from temperate climate colonies. Additionally, overt infections were observed only in the temperate climate. Varroa parasitism and DWV loads in bees from colonies with overt infections were significantly higher than in bees from colonies with covert infections. These results suggest that interactions between climate, V. destructor, and possibly other factors, may play a significant role in the prevalence and levels of DWV in honey bee colonies, as well as in the development of overt infections. Several hypotheses are discussed to explain these results.