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Acaricide, Fungicide and Drug Interactions in Honey Bees (Apis mellifera)
Chemical analysis shows that honey bees (Apis mellifera) and hive products contain many pesticides derived from various sources. The most abundant pesticides are acaricides applied by beekeepers to control Varroa destructor. Beekeepers also apply antimicrobial drugs to control bacterial and microsporidial diseases. Fungicides may enter the hive when applied to nearby flowering crops. Acaricides, antimicrobial drugs and fungicides are not highly toxic to bees alone, but in combination there is potential for heightened toxicity due to interactive effects.
Laboratory bioassays based on mortality rates in adult worker bees demonstrated interactive effects among acaricides, as well as between acaricides and antimicrobial drugs and between acaricides and fungicides. Toxicity of the acaricide tau-fluvalinate increased in combination with other acaricides and most other compounds tested (15 of 17) while amitraz toxicity was mostly unchanged (1 of 15). The sterol biosynthesis inhibiting (SBI) fungicide prochloraz elevated the toxicity of the acaricides tau-fluvalinate, coumaphos and fenpyroximate, likely through inhibition of detoxicative cytochrome P450 monooxygenase activity. Four other SBI fungicides increased the toxicity of tau-fluvalinate in a dose-dependent manner, although possible evidence of P450 induction was observed at the lowest fungicide doses. Non-transitive interactions between some acaricides were observed. Sublethal amitraz pre-treatment increased the toxicity of the three P450-detoxified acaricides, but amitraz toxicity was not changed by sublethal treatment with the same three acaricides. A two-fold change in the toxicity of tau-fluvalinate was observed between years, suggesting a possible change in the genetic composition of the bees tested.
Interactions with acaricides in honey bees are similar to drug interactions in other animals in that P450-mediated detoxication appears to play an important role. Evidence of non-transivity, year-to-year variation and induction of detoxication enzymes indicates that pesticide interactions in bees may be as complex as drug interactions in mammals.
Journal Article
Adjuvants to improve efficacy of miticides in managed honey bee (Apis mellifera) colonies to control Varroa destructor
Beekeepers must manage Varroa destructor mites to maintain colony health. Large-scale beekeepers often use chemical treatments (miticides) to manage this pest. Miticide resistance drives a need for compounds with alternative modes of toxic action that can be used in rotation as part of a Varroa management plan. This research aimed to determine the efficacy of oxalic acid, clove oil, and fenpyroximate when delivered in glycerin soaked in strips and combined with a range of bee-safe adjuvants. Adjuvants are a group of compounds used in plant pesticide applications to increase the spreading and penetration of a pesticide. Laboratory cage trials tested a miticidal active ingredient (oxalic acid, clove oil, or fenpyroximate) and an adjuvant (Ecostep BC-12 ® , Ecostep SE-11 ® , Ecostep AE-13 ® , Ecostep CE-13 ® , or Silwet L-7500 ® ) in glycerin-soaked strips. Field trials evaluated the best performing active ingredient-adjuvant combination from cage trials, oxalic acid combined with Ecostep BC-12 ® adjuvant in glycerin-soaked strips. Neither glycerin control, oxalic acid alone, or oxalic acid with adjuvant caused a significant change in Varroa per 100 bees in field trial year 1, when starting Varroa levels were high (average 11.8 Varroa per 100 bees across all treatment groups). In year 2, when starting Varroa levels were low (average 0.58 Varroa per 100 bees across all treatment groups), Varroa per 100 bees increased 2.6-fold for the glycerin control and 2.8-fold for oxalic acid alone, while a 29% reduction was observed in the oxalic acid with adjuvant treatment. Additionally, mite drop data indicated increased speed for the miticidal effect when an adjuvant is included with oxalic acid. This research informs formulation chemistries for oxalic acid and other miticides to help beekeepers maintain healthy hives.
Journal Article
Real-Time Sequence-Validated Loop-Mediated Isothermal Amplification Assays for Detection of Middle East Respiratory Syndrome Coronavirus (MERS-CoV)
The Middle East respiratory syndrome coronavirus (MERS-CoV), an emerging human coronavirus, causes severe acute respiratory illness with a 35% mortality rate. In light of the recent surge in reported infections we have developed asymmetric five-primer reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for detection of MERS-CoV. Isothermal amplification assays will facilitate the development of portable point-of-care diagnostics that are crucial for management of emerging infections. The RT-LAMP assays are designed to amplify MERS-CoV genomic loci located within the open reading frame (ORF)1a and ORF1b genes and upstream of the E gene. Additionally we applied one-step strand displacement probes (OSD) for real-time sequence-specific verification of LAMP amplicons. Asymmetric amplification effected by incorporating a single loop primer in each assay accelerated the time-to-result of the OSD-RT-LAMP assays. The resulting assays could detect 0.02 to 0.2 plaque forming units (PFU) (5 to 50 PFU/ml) of MERS-CoV in infected cell culture supernatants within 30 to 50 min and did not cross-react with common human respiratory pathogens.
Journal Article
Toxicity of spray adjuvants and tank mix combinations used in almond orchards to adult honey bees (Apis mellifera)
Commercial beekeepers transporting honey bees across the United States to provide almond pollination services have reported honey bee deaths, possibly due to pesticide applications made during crop bloom. Pesticides are often applied as “tank mixes”, or mixtures of fungicides and insecticides combined into a single application. Spray adjuvants are often added to tank mixes to improve the application characteristics of a pesticide and include spreaders, stickers, or surfactants. The goal of this research was to determine toxicity of adjuvants to adult worker honey bees, both when applied alone and in adjuvant-pesticide tank mixtures. Field-relevant combinations of formulated products were applied to 3-day-old adult worker honey bees using a Potter Spray Tower, and mortality was assessed 48 h following exposure. Adjuvants tested included Activator-90, Attach, Choice Weather Master, Cohere, Dyne-Amic, Induce, Kinetic, LI 700, Liberate, Nu-Film P, PHT Latron B-1956, and Surf-90; fungicides tested include Luna Sensation (Fluopyram and Trifloxystrobin), Pristine (Pyraclostrobin and Boscalid), Tilt (Propiconazole), and Vangard (Cyprodinil), and insecticides tested include Altacor (Chlorantraniliprole), Intrepid 2F (Methoxyfenozide), and a positive control Mustang Maxx (Zeta-cypermethrin). Results demonstrated that exposure to some adjuvants causes acute honey bee mortality at near-field application rates, both when applied alone and in combination with pesticides. Some adjuvant-pesticide combinations demonstrated increased toxicity compared with the adjuvant alone, while others demonstrated decreased toxicity. A better understanding of adjuvant and adjuvant-pesticide tank mixture toxicity to honey bees will play a key role in informing “Best Management Practices” for pesticide applicators using spray adjuvants during bloom when honey bee exposure is likely.
Journal Article
Omicron Spike confers enhanced infectivity and interferon resistance to SARS-CoV-2 in human nasal tissue
Omicron emerged following COVID-19 vaccination campaigns, displaced previous SARS-CoV-2 variants of concern worldwide, and gave rise to lineages that continue to spread. Here, we show that Omicron exhibits increased infectivity in primary adult upper airway tissue relative to Delta. Using recombinant forms of SARS-CoV-2 and nasal epithelial cells cultured at the liquid-air interface, we show that mutations unique to Omicron Spike enable enhanced entry into nasal tissue. Unlike earlier variants of SARS-CoV-2, our findings suggest that Omicron enters nasal cells independently of serine transmembrane proteases and instead relies upon metalloproteinases to catalyze membrane fusion. Furthermore, we demonstrate that this entry pathway unlocked by Omicron Spike enables evasion from constitutive and interferon-induced antiviral factors that restrict SARS-CoV-2 entry following attachment. Therefore, the increased transmissibility exhibited by Omicron in humans may be attributed not only to its evasion of vaccine-elicited adaptive immunity, but also to its superior invasion of nasal epithelia and resistance to the cell-intrinsic barriers present therein.
Shi and Li et al. show that SARS-CoV-2 Omicron subvariants have increased capacity to infect primary human nasal tissue using a distinct entry route that depends on matrix metalloproteinases as opposed to TMPRSS2 used by previous variants, which enables evasion from antiviral factors.
Journal Article
Persistence of Severe Acute Respiratory Syndrome Coronavirus 2 in Aerosol Suspensions
We aerosolized severe acute respiratory syndrome coronavirus 2 and determined that its dynamic aerosol efficiency surpassed those of severe acute respiratory syndrome coronavirus and Middle East respiratory syndrome. Although we performed experiment only once across several laboratories, our findings suggest retained infectivity and virion integrity for up to 16 hours in respirable-sized aerosols.
Journal Article
Non-neutralizing antibodies elicited by recombinant Lassa–Rabies vaccine are critical for protection against Lassa fever
Lassa fever (LF), caused by Lassa virus (LASV), is a viral hemorrhagic fever for which no approved vaccine or potent antiviral treatment is available. LF is a WHO priority disease and, together with rabies, a major health burden in West Africa. Here we present the development and characterization of an inactivated recombinant LASV and rabies vaccine candidate (LASSARAB) that expresses a codon-optimized LASV glycoprotein (coGPC) and is adjuvanted by a TLR-4 agonist (GLA-SE). LASSARAB elicits lasting humoral response against LASV and RABV in both mouse and guinea pig models, and it protects both guinea pigs and mice against LF. We also demonstrate a previously unexplored role for non-neutralizing LASV GPC-specific antibodies as a major mechanism of protection by LASSARAB against LF through antibody-dependent cellular functions. Overall, these findings demonstrate an effective inactivated LF vaccine and elucidate a novel humoral correlate of protection for LF.
There is currently no approved vaccine for Lassa fever virus (LASV). Here, Abreu-Mota et al. develop an inactivated, adjuvanted vaccine candidate expressing LASV glycoprotein (GPC) in a rabies virus vector, and show that non-neutralizing LASV GPC-specific antibodies are a major mechanism of protection.
Journal Article
Adjuvants to improve efficacy of miticides in managed honey bee
Beekeepers must manage Varroa destructor mites to maintain colony health. Large-scale beekeepers often use chemical treatments (miticides) to manage this pest. Miticide resistance drives a need for compounds with alternative modes of toxic action that can be used in rotation as part of a Varroa management plan. This research aimed to determine the efficacy of oxalic acid, clove oil, and fenpyroximate when delivered in glycerin soaked in strips and combined with a range of bee-safe adjuvants. Adjuvants are a group of compounds used in plant pesticide applications to increase the spreading and penetration of a pesticide. Laboratory cage trials tested a miticidal active ingredient (oxalic acid, clove oil, or fenpyroximate) and an adjuvant (Ecostep BC-12.sup.®, Ecostep SE-11.sup.®, Ecostep AE-13.sup.®, Ecostep CE-13.sup.®, or Silwet L-7500.sup.®) in glycerin-soaked strips. Field trials evaluated the best performing active ingredient-adjuvant combination from cage trials, oxalic acid combined with Ecostep BC-12.sup.® adjuvant in glycerin-soaked strips. Neither glycerin control, oxalic acid alone, or oxalic acid with adjuvant caused a significant change in Varroa per 100 bees in field trial year 1, when starting Varroa levels were high (average 11.8 Varroa per 100 bees across all treatment groups). In year 2, when starting Varroa levels were low (average 0.58 Varroa per 100 bees across all treatment groups), Varroa per 100 bees increased 2.6-fold for the glycerin control and 2.8-fold for oxalic acid alone, while a 29% reduction was observed in the oxalic acid with adjuvant treatment. Additionally, mite drop data indicated increased speed for the miticidal effect when an adjuvant is included with oxalic acid. This research informs formulation chemistries for oxalic acid and other miticides to help beekeepers maintain healthy hives.
Journal Article
Ecologically Appropriate Xenobiotics Induce Cytochrome P450s in Apis mellifera
Honey bees are exposed to phytochemicals through the nectar, pollen and propolis consumed to sustain the colony. They may also encounter mycotoxins produced by Aspergillus fungi infesting pollen in beebread. Moreover, bees are exposed to agricultural pesticides, particularly in-hive acaricides used against the parasite Varroa destructor. They cope with these and other xenobiotics primarily through enzymatic detoxificative processes, but the regulation of detoxificative enzymes in honey bees remains largely unexplored.
We used several approaches to ascertain effects of dietary toxins on bee susceptibility to synthetic and natural xenobiotics, including the acaricide tau-fluvalinate, the agricultural pesticide imidacloprid, and the naturally occurring mycotoxin aflatoxin. We administered potential inducers of cytochrome P450 enzymes, the principal biochemical system for Phase 1 detoxification in insects, to investigate how detoxification is regulated. The drug phenobarbital induces P450s in many insects, yet feeding bees with phenobarbital had no effect on the toxicity of tau-fluvalinate, a pesticide known to be detoxified by bee P450s. Similarly, no P450 induction, as measured by tau-fluvalinate tolerance, occurred in bees fed xanthotoxin, salicylic acid, or indole-3-carbinol, all of which induce P450s in other insects. Only quercetin, a common pollen and honey constituent, reduced tau-fluvalinate toxicity. In microarray comparisons no change in detoxificative gene expression was detected in phenobarbital-treated bees. However, northern blot analyses of guts of bees fed extracts of honey, pollen and propolis showed elevated expression of three CYP6AS P450 genes. Diet did not influence tau-fluvalinate or imidacloprid toxicity in bioassays; however, aflatoxin toxicity was higher in bees consuming sucrose or high-fructose corn syrup than in bees consuming honey.
These results suggest that regulation of honey bee P450s is tuned to chemicals occurring naturally in the hive environment and that, in terms of toxicological capacity, a diet of sugar is not equivalent to a diet of honey.
Journal Article