Abstract
The stingless bee Frieseomelitta varia Lepeletier 1836 (Hymenoptera: Apidae) is an essential pollinator in natural and agricultural ecosystems in the Neotropical region. However, these bees may be exposed to pesticides during foraging, which can affect both individuals and their colonies. One example comes from the use of pyraclostrobin (a fungicide) and thiamethoxam (an insecticide) for pest control in pepper crops, which F. varia visits. This study aimed to evaluate the isolated and combined sublethal effects of thiamethoxam (TMX) (0.000543 ng a.i./µL) and pyraclostrobin (PYR) (1.5 ng i.a./µL) on the morphology of the midgut and Malpighian tubules of F. varia workers. Results showed that both pesticides, regardless of the exposure time (through feeding during 48 h or 96 h), disturbed the morphology of the analyzed organs. Specifically, F. varia exposed orally to sublethal concentrations of thiamethoxam and pyraclostrobin, either alone or in combination, exhibited a higher rate of damage to the midgut (e.g., vacuolization, apocrine secretion, and cellular elimination) compared to the bees in the control groups, both after 48 h and 96 h of exposure. In Malpighian tubules, vacuolation is the only damage present. As the observed morphological alterations likely compromise the excretion and absorption functions, exposure to pyraclostrobin and thiamethoxam may lead to disturbances at both the individual and colony levels. These results highlight the urgent need for a future reassessment of the safety of fungicides and insecticides regarding their potential effects on bee populations.
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References
ADAPAR (2023) Agência Nacional de Defesa Agropecuária do Paraná, Curitiba. Available in: http://celepar07web.pr.gov.br/agrotoxicos/pesquisar.asp. Accessed 22 Oct 2023. Accessed 26 Jun 2007
Bartlett DW, Clough JM, Godwin JR, Hall AA, Hamer M, Parr‐Dobrzanski B (2002) The strobilurin fungicides. Pest Manag Sci 58:649–662. https://doi.org/10.1002/ps.520
Batista AC, Domingues CEC, Costa MJ, Silva-Zacarin ECM (2020a) Is a strobilurin fungicide capable of inducing histopathological effects on the midgut and Malpighian tubules of honey bees? J Apic Res 59:834–843. https://doi.org/10.1080/00218839.2020.1724678
Batista NR, Farder-Gomes CF, Nocelli RCF, Antonialli-Junior WF (2023) Effects of chronic exposure to sublethal doses of neonicotinoids in the social wasp Polybia paulista: survival, mobility, and histopathology. Sci Total Environ 904:166823. https://doi.org/10.1016/j.scitotenv.2023.166823
Botina LL, Barbosa WF, Martins GF (2024) Toxicological assessments of agrochemicals in stingless bees in Brazil: a systematic review. Neotrop. Entomol 1:10. https://doi.org/10.1007/s13744-024-01132-x
BPBES/REBIPP (2019) Relatório temático sobre Polinização, Polinizadores e Produção de 50 Alimentos no Brasil. Campinas, São Paulo.
Camargo JMF, Pedro SRM, Melo GAR (2023) Meliponini Lepeletier, 1836. In Moure, J.S., Urban, D, Melo GAR (Orgs). Catalogue of Bees (Hymenoptera, Apoidea) in the Neotropical Region - online version. Available at http://www.moure.cria.org.br/catalogue. Accessed 22 Oct 2023.
Campos VAC, Santos Júnior HMD, Oliveira DF, Carvalho HWPD, Machado ART, Tirelli AA (2011) Antibacterial activity of propolis produced by Frieseomelitta varia. Ciência e Agrotecnologia 35:1043–1049. https://doi.org/10.1590/S1413-70542011000600002
Carneiro LS, Santos CG, de Resende MTCS, de Souza DLL, dos Santos Souza D, da Cruz Souza AM, Motta JVDO, Nere PHA, Oliveira AHD Serrão JE (2023) Effects of the insecticide imidacloprid on the post-embryonic development of the honey bee Apis mellifera (Hymenoptera: Apidae). Sci Total Environ 167278. https://doi.org/10.1016/j.scitotenv.2023.167278
Catae AF, Roat TC, De Oliveira RA, Ferreira Nocelli RC, Malaspina O (2014) Cytotoxic effects of thiamethoxam in the midgut and malpighian tubules of Africanized Apis mellifera (Hymenoptera: Apidae). Microsc Res Tech 77:274–281. https://doi.org/10.1002/jemt.22339
Colwell MJ, Williams GR, Evans RC, Shutler D (2017) Honey bee‐collected pollen in agro‐ecosystems reveals diet diversity, diet quality, and pesticide exposure. Ecol Evol 7:7243–7253. https://doi.org/10.1002/ece3.3178
Correia SC, Resende R, Moreira PI, Pereira CM (2015) Alzheimer’s disease-related misfolded proteins and dysfunctional organelles on autophagy menu. DNA Cell Biol 34:261–273. https://doi.org/10.1089/dna.2014.2757
da Costa Domingues CE, Inoue LVB, da Silva-Zacarin ECM, Malaspina O (2020a) Fungicide pyraclostrobin affects midgut morphophysiology and reduces survival of Brazilian native stingless bee Melipona scutellaris. Ecotoxicol Environ Saf 206:111395. https://doi.org/10.1016/j.ecoenv.2020.111395
da Costa Domingues CE, Inoue LVB, da Silva-Zacarin ECM, Malaspina O (2020b) Foragers of Africanized honeybee are more sensitive to fungicide pyraclostrobin than newly emerged bees. Environ Pollut 266:115267. https://doi.org/10.1016/j.envpol.2020.115267
David A, Botías C, Abdul-Sada A, Goulson D, Hill EM (2015) Sensitive determination of mixtures of neonicotinoid and fungicide residues in pollen and single bumblebees using a scaled down QuEChERS method for exposure assessment. Anal Bioanal Chem 407:8151–8162. https://doi.org/10.1007/s00216-015-8986-6
de Assis JC, Domingues CEC, Tadei R, da Silva CI, Lima HMS, Decio P, Silva-Zacarin EC (2022) Sublethal doses of imidacloprid and pyraclostrobin impair fat body of solitary bee Tetrapedia diversipes (Klug, 1810). Environ Pollut 304:119140. https://doi.org/10.1016/j.envpol.2022.119140
dos Santos-Junior VC, Martínez LC, Plata-Rueda A, Bozdoğan H, Zanuncio JC, Serrão JE (2019) Exposure to spinosad induces histopathological and cytotoxic effects on the salivary complex of the non-target predator Podisus nigrispinus. Chemosphere 225:688–695. https://doi.org/10.1016/j.chemosphere.2019.03.105
Decourtye A, Devillers J, Cluzeau S, Charreton M, Pham-Delègue MH (2004) Effects of imidacloprid and deltamethrin on associative learning in honeybees under semi-field and laboratory conditions. Ecotoxicol Environ Saf 57:410–419. https://doi.org/10.1016/j.ecoenv.2003.08.001
Dong J, Huang M, Guo H, Zhang J, Tan X, Wang D (2023) Ternary mixture of azoxystrobin, boscalid and pyraclostrobin disrupts the gut microbiota and metabolic balance of honeybees (Apis cerana cerana). Int J Mol Sci 24:5354. https://doi.org/10.3390/ijms24065354
Farder-Gomes CF, Fernandes KM, Bernardes RC, Bastos DSS, Martins GF, Serrão JE (2021) Acute exposure to fipronil induces oxidative stress, apoptosis and impairs epithelial homeostasis in the midgut of the stingless bee Partamona helleri Friese (Hymenoptera: Apidae). Sci Total Environ 774:145679. https://doi.org/10.1016/j.scitotenv.2021.145679
Farder-Gomes CF, Fernandes KM, Bernardes RC, Martins GF, Serrão JE (2022) Fipronil exposure compromises respiration and damages the Malpighian tubules of the stingless bee Partamona helleri Friese (Hymenoptera: Apidae). Environ Sci Pollut Res 29:88101–88108. https://doi.org/10.1007/s11356-022-21858-8
Farder-Gomes CF, de Oliveira MA, Malaspina O, Nocelli RFC (2024a) Exposure of the stingless bee Melipona scutellaris to imidacloprid, pyraclostrobin, and glyphosate, alone and in combination, impair its walking activity and fat body morphology and physiology. Environ Pollut 123783. https://doi.org/10.1016/j.envpol.2024.123783
Farder-Gomes CF, Grella TC, Malaspina O, Nocelli RFC (2024b) Exposure to sublethal concentrations of imidacloprid, pyraclostrobin, and glyphosate harm the behavior and fat body cells of the stingless bee Scaptotrigona postica. Sci Total Environ 907:168072. https://doi.org/10.1016/j.scitotenv.2023.168072
Farder-Gomes CF, Miranda FR, Fernandes KM, Bernardes RC, Bastos DSS, de Oliveira LL, Martins GF, Serrão JE (2024c) Exposure to low-concentration fipronil impairs survival, behavior, midgut morphology and physiology of Aedes aegypti larvae. Chemosphere 358:142240. https://doi.org/10.1016/j.chemosphere.2024.142240
Fiaz M, Martínez LC, Plata-Rueda A, Gonçalves WG, de Souza DLL, Cossolin JFS, Carvalho PEGR, Martins GF, Serrão JE (2019) Pyriproxyfen, a juvenile hormone analog, damages midgut cells and interferes with behaviors of Aedes aegypti larvae. PeerJ 7:e7489. https://doi.org/10.7717/peerj.7489
Gong Y, Diao Q (2017) Current knowledge of detoxification mechanisms of xenobiotic in honey bees. Ecotoxicol 26:1–12. https://doi.org/10.1007/s10646-016-1742-7
Grella TC, Soares-Lima HM, Malaspina O, Nocelli RCF (2019) Semi-quantitative analysis of morphological changes in bee tissues: a toxicological approach. Chemosphere 236:124255. https://doi.org/10.1016/j.chemosphere.2019.06.225
Han W, Ye Z, Gu Y, Zhong Y, Gao J, Zhao S, Wang S (2023) Gut microbiota composition and gene expression changes induced in the Apis cerana exposed to acetamiprid and difenoconazole at environmentally realistic concentrations alone or combined. Front Physiol 14:1174236. https://doi.org/10.3389/fphys.2023.1174236
Huang M, Dong J, Yang S, **ao M, Guo H, Zhang J, Wang D (2023) Ecotoxicological effects of common fungicides on the eastern honeybee Apis cerana cerana (Hymenoptera). Sci Total Environ 868:161637. https://doi.org/10.1016/j.scitotenv.2023.161637
ICMBio (2018) Livro vermelho de fauna brasileira ameaçada de extinção. Instituto Chico Mendes de Conservação da Biodiversidade, Brasília.
Inoue LV, Domingues CE, Gregorc A, Silva-Zacarin EC, Malaspina O (2022) Harmful effects of pyraclostrobin on the fat body and pericardial cells of foragers of Africanized honey bee. Toxics 10:530. https://doi.org/10.3390/toxics10090530
Ishaaya I, Barazani A, Kontsedalov S, Horowitz AR (2007) Insecticides with novel modes of action: mechanism, selectivity and cross‐resistance. Entomol Res 37:148–152. https://doi.org/10.1111/j.1748-5967.2007.00104.x
Kevan PG, Viana BF (2003) The global decline of pollination services. Biodivers 4:3–8. https://doi.org/10.1080/14888386.2003.9712703
Kremen C, Williams NM, Thorp RW (2002) Crop pollination from native bees at risk from agricultural intensification. Proc. Natl Acad Sci USA 99:16812–16816. https://doi.org/10.1073/pnas.262413599
Liu J, Zhang X, Wu H, Gao Y, Silver K, Ma E, Zhang J, Zhu KY (2018) Comparisons of microsomal cytochrome P450 content and enzymatic activity towards selected model substrates and insecticides in different tissues from the migratory locust (Locusta migratoria). Chemosphere 208:366–373. https://doi.org/10.1016/j.chemosphere.2018.05.179
Liu Y, Levine B (2015) Autosis and autophagic cell death: the dark side of autophagy. Cell Death Differ 22:367–376. https://doi.org/10.1038/cdd.2014.143
Li H, Cao F, Zhao F, Yang Y, Teng M, Wang C, Qiu L (2018) Developmental toxicity, oxidative stress and immunotoxicity induced by three strobilurins (pyraclostrobin, trifloxystrobin and picoxystrobin) in zebrafish embryos. Chemosphere 207:781–790. https://doi.org/10.1016/j.chemosphere.2018.05.146
Li W, Lv L, Wang Y, Zhu YC (2023) Mixture effects of thiamethoxam and seven pesticides with different modes of action on honey bees (Apis mellifera). Sci Rep 13:2679. https://doi.org/10.1038/s41598-023-29837-w
Lv L, Li W, Li X, Wang D, Weng H, Zhu YC, Wang Y (2023) Mixture toxic effects of thiacloprid and cyproconazole on honey bees (Apis mellifera L.). Sci Total Environ 870:161700. https://doi.org/10.1016/j.scitotenv.2023.161700
Martínez LC, Plata-Rueda A, da Silva Neves G, Gonçalves WG, Zanuncio JC, Bozdoğan H, Serrão JE (2018) Permethrin induces histological and cytological changes in the midgut of the predatory bug, Podisus nigrispinus. Chemosphere 212:629–637. https://doi.org/10.1016/j.chemosphere.2018.08.134
Michener CD (2013) The meliponini, pot-honey, a legacy of stingless bees. Springer, New York
Miotelo L, Dos Reis ALM, Rosa-Fontana A, da Silva Pachú JK, Malaquias JB, Malaspina O, Roat TC (2022) A food-ingested sublethal concentration of thiamethoxam has harmful effects on the stingless bee Melipona scutellaris. Chemosphere 288:132461. https://doi.org/10.1016/j.chemosphere.2021.132461
Miotelo L, Dos Reis ALM, Malaquias JB, Malaspina O, Roat TC (2021) Apis mellifera and Melipona scutellaris exhibit differential sensitivity to thiamethoxam. Environ Pollut 268:115770. https://doi.org/10.1016/j.envpol.2020.115770
Meech R, Miners JO, Lewis BC, Mackenzie PI (2012) The glycosidation of xenobiotics and endogenous compounds: versatility and redundancy in the UDP glycosyltransferase superfamily. Pharmacol Therapeut 134:200–218. https://doi.org/10.1016/j.pharmthera.2012.01.009
Moore MN, Allen JI, Somerfield PJ (2006) Autophagy: role in surviving environmental stress. Mar. Environ Res 62:S420–S425. https://doi.org/10.1016/j.marenvres.2006.04.055
Nocelli RC, Cintra-Socolowski P, Roat TC, Silva-Zacarin EC, Malaspina O (2016) Comparative physiology of Malpighian tubules: form and function. Open Access Insect Physiol 13–23. https://doi.org/10.2147/OAIP.S72060
OECD (Organization for Economic Co-operation and Development) (1998) OECD Guidelines for the Testing of Chemicals, Honeybees, Acute Oral Toxicity Test n°213
Oliveira CR, Domingues CE, de Melo NF, Roat TC, Malaspina O, Jones-Costa M, Silva-Zacarin, Fraceto LF (2019) Nanopesticide based on botanical insecticide pyrethrum and its potential effects on honeybees. Chemosphere 236:124282. https://doi.org/10.1016/j.chemosphere.2019.07.013
Papach A, Fortini D, Grateau S, Aupinel P, Richard FJ (2017) Larval exposure to thiamethoxam and American foulbrood: effects on mortality and cognition in the honey bee Apis mellifera. J Apic Res 56:475–486. https://doi.org/10.1080/00218839.2017.1332541
Pedro SR(2014) The stingless bee fauna in Brazil (Hymenoptera: Apidae) Sociobiology 61:348–354. https://doi.org/10.13102/sociobiology.v61i4.348-354
Pettis JS, Lichtenberg EM, Andree M, Stitzinger J, Rose R, Vanengelsdorp D (2013) Crop pollination exposes honey bees to pesticides which alters their susceptibility to the gut pathogen Nosema ceranae. PloS ONE 8:e70182. https://doi.org/10.1371/journal.pone.0070182
Piechowicz B, Szpyrka E, Zaręba L, Podbielska M, Grodzicki P (2018) Transfer of the active ingredients of some plant protection products from raspberry plants to beehives. Arch Environ Contam Toxicol 75:45–58. https://doi.org/10.1007/s00244-017-0488-4
Potts SG, Biesmeijer JC, Kremen C, Neumann P, Schweiger O, Kunin WE (2010) Global pollinator declines: trends, impacts and drivers. Trends Ecol Evol 25:345–353. https://doi.org/10.1016/j.tree.2010.01.007
Potts SG, Imperatriz-Fonseca V, Ngo HT, Aizen MA, Biesmeijer JC, Breeze TD, Dicks LV, Garibaldi LA, Hill R, Settele J, Vanbergen AJ (2016) Safeguarding pollinators and their values to human well-being. Nature 540:220–229. https://doi.org/10.1038/nature20588
Ryter SW, Cloonan SM, Choi AM (2013) Autophagy: a critical regulator of cellular metabolism and homeostasis. Mol Cells 36:7–16. https://doi.org/10.1007/s10059-013-0140-8
Roat TC, dos Santos-Pinto JRA, Miotelo L, de Souza CL, Palma MS, Malaspina O (2020) Using a toxicoproteomic approach to investigate the effects of thiamethoxam into the brain of Apis mellifera. Chemosphere 258:127362. https://doi.org/10.1016/j.chemosphere.2020.127362
Slaa EJ, Chaves LAS, Malagodi-Braga KS, Hofstede FE (2006) Stingless bees in applied pollination: practice and perspectives. Apidologie 37(2):293–315. https://doi.org/10.1051/apido:2006022
Schmuck R, Stadler T, Schmidt HW (2003) Field relevance of a synergistic effect observed in the laboratory between an EBI fungicide and a chloronicotinyl insecticide in the honeybee (Apis mellifera L, Hymenoptera). Pest Manag Sci 59:279–286. https://doi.org/10.1002/ps.626
Schuhmann A, Schmid AP, Manzer S, Schulte J, Scheiner R (2022) Interaction of insecticides and fungicides in bees. Front Insect Sci 1. https://doi.org/10.3389/finsc.2021.808335
Tadei R, Domingues CEC, Malaquias JB, Camilo EV, Malaspina O, Silva-Zacarin EC (2019) Late effect of larval co-exposure to the insecticide clothianidin and fungicide pyraclostrobin in Africanized Apis mellifera. Sci Rep 9:3277. https://doi.org/10.1038/s41598-019-39383-z
Thompson HM, Fryday SL, Harkin S, Milner S (2014) Potential impacts of synergism in honeybees (Apis mellifera) of exposure to neonicotinoids and sprayed fungicides in crops. Apidologie 45:545–553. https://doi.org/10.1007/s13592-014-0273-6
Tosi S, Costa C, Vesco U, Quaglia G, Guido G (2018) A 3-year survey of Italian honey bee-collected pollen reveals widespread contamination by agricultural pesticides. Sci Total Environ 615:208–218. https://doi.org/10.1016/j.scitotenv.2017.09.226
Vicentini CB, Romagnoli C, Andreotti E, Mares D (2007) Synthetic pyrazole derivatives as growth inhibitors of some phytopathogenic fungi. J Agric Food Chem 55:10331–10338. https://doi.org/10.1021/jf072077d
Wernecke A, Frommberger M, Forster R, Pistorius J (2019) Lethal effects of various tank mixtures including insecticides, fungicides and fertilizers on honey bees under laboratory, semi-field and field conditions. J Food Saf 14:239–249. https://doi.org/10.1007/s00003-019-01233-5
Xu X, Wang X, Yang Y, Ares I, Martínez M, Lopez-Torres B, Martínez-Larrañaga MR, Wang X, Anadón A, Martinez MA (2022) Neonicotinoids: mechanisms of systemic toxicity based on oxidative stress-mitochondrial damage. Arch Toxicol 96:1493–1520. https://doi.org/10.1007/s00204-022-03267-5
Yoder JA, Jajack AJ, Rosselot AE, Smith TJ, Yerke MC, Sammataro D (2013) Fungicide contamination reduces beneficial fungi in bee bread based on an area-wide field study in honey bee, Apis mellifera, colonies. J Toxicol Environ Health Part A 76:587–600. https://doi.org/10.1080/15287394.2013.798846
Zioga E, White B, Stout JC (2023) Honey bees and bumble bees may be exposed to pesticides differently when foraging on agricultural areas. Sci Total Environ 896:166214. https://doi.org/10.1016/j.scitotenv.2023.166214
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The authors are thankful to Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001 and the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (grant numbers: 2017/21097-3; 2021/09996-8).
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Conceptualization: Jaqueline Aparecida da Silva and Cliver Fernandes Farder-Gomes; methodology: Jaqueline Aparecida da Silva and Cliver Fernandes Farder-Gomes; formal analysis and investigation: Jaqueline Aparecida da Silva and Cliver Fernandes Farder-Gomes; writing—original draft preparation: Jaqueline Aparecida da Silva; writing—review and editing: Jaqueline Aparecida da Silva, Cliver Fernandes Farder-Gomes, Angel Roberto Barchuk, and Roberta Cornélio Ferreira Nocelli; funding acquisition: Roberta Cornélio Ferreira Nocelli and Osmar Malaspina; resources: Roberta Cornélio Ferreira Nocelli and Osmar Malaspina; supervision: Roberta Cornélio Ferreira Nocelli.
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da Silva, J.A., Farder-Gomes, C.F., Barchuk, A.R. et al. Sublethal exposure to thiamethoxam and pyraclostrobin affects the midgut and Malpighian tubules of the stingless bee Frieseomelitta varia (Hymenoptera: Apidae: Meliponini). Ecotoxicology (2024). https://doi.org/10.1007/s10646-024-02786-4
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DOI: https://doi.org/10.1007/s10646-024-02786-4