Acaricide, Fungicide and Drug Interactions in Honey Bees (Apis mellifera)

Background 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. Methodology/Principal Findings 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. Conclusions/Significance 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.

[1]  R. Downer,et al.  Interaction of formamidine pesticides with insect neural octopamine receptors: Effects on radioligand binding and cyclic AMP production , 1990 .

[2]  Yves-Jacques Schneider,et al.  CYP1A1 induction and CYP3A4 inhibition by the fungicide imazalil in the human intestinal Caco-2 cells-comparison with other conazole pesticides. , 2009, Toxicology letters.

[3]  S. Wilkins,et al.  Assessment of the synergy and repellency of pyrethroid/fungicide mixtures , 2003 .

[4]  A. Imdorf,et al.  ACARICIDE RESIDUES IN SOME BEE PRODUCTS , 1998 .

[5]  E. Rademacher,et al.  Oxalic acid for the control of varroosis in honey bee colonies – a review , 2006 .

[6]  Ming-Yie Liu,et al.  Mechanism of formamidine synergism of pyrethroids , 1992 .

[7]  Jay D. Evans,et al.  Colony Collapse Disorder: A Descriptive Study , 2009, PloS one.

[8]  J. Frazier,et al.  "Entombed Pollen": A new condition in honey bee colonies associated with increased risk of colony mortality. , 2009, Journal of invertebrate pathology.

[9]  K. Wallner Varroacides and their residues in bee products , 1999 .

[10]  S. Bogdanov Beeswax: quality issues today , 2004 .

[11]  M. Berenbaum,et al.  Quercetin-metabolizing CYP6AS enzymes of the pollinator Apis mellifera (Hymenoptera: Apidae). , 2009, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[12]  A. Mara,et al.  A chemical and genetic approach to the mode of action of fumagillin. , 2006, Chemistry & biology.

[13]  M. Berenbaum,et al.  A deficit of detoxification enzymes: pesticide sensitivity and environmental response in the honeybee , 2006, Insect molecular biology.

[14]  E. Genersch,et al.  Varroosis – the Ongoing Crisis in Bee Keeping , 2008, Journal für Verbraucherschutz und Lebensmittelsicherheit.

[15]  S. Imamura,et al.  Effect of antibiotics on the generation of reactive oxygen species. , 1986, The Journal of investigative dermatology.

[16]  J. Nowacki,et al.  Residues of captan (contact) and difenoconazole (systemic) fungicides in bee products from an apple orchard , 2000 .

[17]  Måns Ehrenberg,et al.  The mechanism of action of macrolides, lincosamides and streptogramin B reveals the nascent peptide exit path in the ribosome. , 2003, Journal of molecular biology.

[18]  C. O. Knowles,et al.  Amitraz effect on the pharmacokinetics of permethrin in Helicoverpa zea (Lepidoptera: Noctuidae) , 1995 .

[19]  J. Gaddum Probit Analysis , 1948, Nature.

[20]  H. Nojiri,et al.  Oxidative damages in isolated rat hepatocytes treated with the organochlorine fungicides captan, dichlofluanid and chlorothalonil. , 2004, Toxicology.

[21]  W. Ritter,et al.  Varroa mites and honey bee health: can Varroa explain part of the colony losses? , 2010, Apidologie.

[22]  J. Faucon,et al.  Pesticide residues in beeswax samples collected from honey bee colonies (Apis mellifera L.) in France. , 2007, Pest management science.

[23]  J. Hothersall,et al.  Crystal and microparticle effects on MDCK cell superoxide production: oxalate-specific mitochondrial membrane potential changes. , 2005, Free radical biology & medicine.

[24]  J. Jonassen,et al.  Mitochondrial dysfunction is a primary event in renal cell oxalate toxicity. , 2004, Kidney international.

[25]  L. Belzunces,et al.  Joint actions of deltamethrin and azole fungicides on honey bee thermoregulation , 1998, Neuroscience Letters.

[26]  David J. Hawthorne,et al.  Killing Them with Kindness? In-Hive Medications May Inhibit Xenobiotic Efflux Transporters and Endanger Honey Bees , 2011, PloS one.

[27]  Reed M. Johnson,et al.  Pesticides and honey bee toxicity — USA , 2010, Apidologie.

[28]  Henry S. Pollock,et al.  Synergistic Interactions Between In-Hive Miticides in Apis mellifera , 2009, Journal of economic entomology.

[29]  P. Rosenkranz,et al.  Biology and control of Varroa destructor. , 2010, Journal of invertebrate pathology.

[30]  M. Scharf,et al.  Bioassays with Arthropods , 2008 .

[31]  D. Weaver,et al.  Effects of Fluvalinate and Coumaphos on Queen Honey Bees (Hymenoptera: Apidae) in Two Commercial Queen Rearing Operations , 2002, Journal of economic entomology.

[32]  C. I. Bliss THE TOXICITY OF POISONS APPLIED JOINTLY1 , 1939 .

[33]  N. Savin,et al.  A Critical Evaluation of Bioassay in Insecticide Research: Likelihood Ratio Tests of Dose-Mortality Regression , 1977 .

[34]  J. Frazier,et al.  High Levels of Miticides and Agrochemicals in North American Apiaries: Implications for Honey Bee Health , 2010, PloS one.

[35]  M. Lodesani,et al.  Ineffectiveness of Apistan® treatment against the mite Varroa jacobsoni Oud in several districts of Lombardy (Italy) , 1995 .

[36]  Gary W Miller,et al.  Mechanism of toxicity of pesticides acting at complex I: relevance to environmental etiologies of Parkinson’s disease , 2007, Journal of neurochemistry.

[37]  D. Bhatnagar,et al.  Lipid peroxidative damage on pyrethroid exposure and alterations in antioxidant status in rat erythrocytes: a possible involvement of reactive oxygen species. , 1999, Toxicology letters.

[38]  M. Smart,et al.  Honey bees (Apis mellifera) reared in brood combs containing high levels of pesticide residues exhibit increased susceptibility to Nosema (Microsporidia) infection. , 2012, Journal of invertebrate pathology.

[39]  F. Paumgartten,et al.  In vitro inhibition of liver monooxygenases by beta-ionone, 1,8-cineole, (-)-menthol and terpineol. , 1999, Toxicology.

[40]  S. Zeggane,et al.  Acaricide residues in honey and wax after treatment of honey bee colonies with Apivar® or Asuntol®50 , 2007, Apidologie.

[41]  M. Vighi,et al.  Coumaphos Distribution in the Hive Ecosystem: Case Study for Modeling Applications , 2004, Ecotoxicology.

[42]  P. Batterham,et al.  Piperonyl butoxide induces the expression of cytochrome P450 and glutathione S-transferase genes in Drosophila melanogaster. , 2007, Pest management science.

[43]  Q. Wang,et al.  Effects of sublethal concentrations of bifenthrin and deltamethrin on fecundity, growth, and development of the honeybee Apis mellifera ligustica , 2010, Environmental toxicology and chemistry.

[44]  B. Gallo,et al.  Study of the degradation products of bromopropylate, chlordimeform, coumaphos, cymiazole, flumethrin and tau-fluvalinate in aqueous media. , 2000, Talanta.

[45]  S. Bogdanov Contaminants of bee products , 2006 .

[46]  M. Berenbaum,et al.  Mediation of pyrethroid insecticide toxicity to honey bees (Hymenoptera: Apidae) by cytochrome P450 monooxygenases. , 2006, Journal of economic entomology.

[47]  A. Gregorc,et al.  HISTOCHEMICAL CHARACTERIZATION OF CELL DEATH IN HONEYBEE LARVAE MIDGUT AFTER TREATMENT WITH PAENIBACILLUS LARVAE, AMITRAZ AND OXYTETRACYCLINE , 2000, Cell biology international.

[48]  A. Imdorf,et al.  Residues in wax and honey after Apilife VAR® treatment , 1998 .

[49]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[50]  B. Siegfried,et al.  Acute contact toxicity of oxalic acid to Varroa destructor (Acari: Varroidae) and their Apis mellifera (Hymenoptera: Apidae) hosts in laboratory bioassays. , 2006, Journal of economic entomology.

[51]  Henry S. Pollock,et al.  Ecologically Appropriate Xenobiotics Induce Cytochrome P450s in Apis mellifera , 2012, PloS one.

[52]  Simon J. Yu The Toxicology and Biochemistry of Insecticides , 2008 .

[53]  M. Uchida,et al.  Effect of a new acaricide, fenpyroximate, on energy metabolism and mitochondrial morphology in adult female Tetranychus urticae (two-spotted spider mite) , 1992 .

[54]  L. Belzunces,et al.  Evidence of synergy between prochloraz and deltamethrin in apis mellifera L.: a convenient biological approach , 1992 .

[55]  P. Elzen,et al.  Detection of coumaphos resistance in Varroa destructor in Florida. , 2002 .

[56]  B. Gallo,et al.  Kinetics and mechanism of amitraz hydrolysis in aqueous media by HPLC and GC-MS. , 1999, Talanta.

[57]  Richard J. Gill,et al.  Combined pesticide exposure severely affects individual- and colony-level traits in bees , 2012, Nature.

[58]  C. Sanders,et al.  Toxicity of antibacterial agents: mechanism of action on mammalian cells. , 1979, Annual review of pharmacology and toxicology.

[59]  J. M. Graham,et al.  The hive and the honey bee. , 1992 .

[60]  J. Marioli,et al.  The concentration effect of selected acaricides present in beeswax foundation on the survival of Apis mellifera colonies , 2012 .

[61]  C. Walker,et al.  Mechanism of synergism between the pyrethroid insecticide λ-cyhalothrin and the imidazole fungicide prochloraz, in the honeybee (Apis mellifera L.) , 1995 .

[62]  C. Vale,et al.  Allosteric positive interaction of thymol with the GABAA receptor in primary cultures of mouse cortical neurons , 2006, Neuropharmacology.

[63]  D. Danehower,et al.  Analysis of chlorothalonil and degradation products in soil and water by GC/MS and LC/MS. , 2008, Chemosphere.

[64]  C. Peng,et al.  Effects of Selected Fungicides on Growth and Development of Larval Honey Bees, Apis mellifera L. (Hymenoptera: Apidae) , 2004 .

[65]  D. P. Highwood,et al.  Fungicide resistance action committee. , 1990 .

[66]  T. Anke The antifungal strobilurins and their possible ecological role , 1995 .

[67]  E. Genersch,et al.  The German bee monitoring project: a long term study to understand periodically high winter losses of honey bee colonies , 2010, Apidologie.

[68]  J. Wu,et al.  Sub-Lethal Effects of Pesticide Residues in Brood Comb on Worker Honey Bee (Apis mellifera) Development and Longevity , 2011, PloS one.

[69]  M. Berenbaum,et al.  CYP9Q-mediated detoxification of acaricides in the honey bee (Apis mellifera) , 2011, Proceedings of the National Academy of Sciences.

[70]  A. Herrera,et al.  Evaluation of residues of essential oil components in honey after different anti-varroa treatments. , 2005, Journal of agricultural and food chemistry.

[71]  T. Seeley The Wisdom of the Hive: The Social Physiology of Honey Bee Colonies , 1995 .