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<front>
<journal-meta>
<journal-id journal-id-type="publisher-id">Front. Bee Sci.</journal-id>
<journal-title>Frontiers in Bee Science</journal-title>
<abbrev-journal-title abbrev-type="pubmed">Front. Bee Sci.</abbrev-journal-title>
<issn pub-type="epub">2813-5911</issn>
<publisher>
<publisher-name>Frontiers Media S.A.</publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="doi">10.3389/frbee.2025.1657493</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Bee Science</subject>
<subj-group>
<subject>Systematic Review</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group>
<article-title>The effect of neonicotinoids on bumblebees (<italic>Bombus</italic> spp.): a systematic review</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname>Dennis</surname>
<given-names>Daisy J.</given-names>
</name>
<uri xlink:href="https://loop.frontiersin.org/people/3191175/overview"/>
<role content-type="https://credit.niso.org/contributor-roles/methodology/"/>
<role content-type="https://credit.niso.org/contributor-roles/conceptualization/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-original-draft/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-review-editing/"/>
<role content-type="https://credit.niso.org/contributor-roles/investigation/"/>
<role content-type="https://credit.niso.org/contributor-roles/visualization/"/>
<role content-type="https://credit.niso.org/contributor-roles/formal-analysis/"/>
<role content-type="https://credit.niso.org/contributor-roles/data-curation/"/>
</contrib>
<contrib contrib-type="author" corresp="yes">
<name>
<surname>Gibbs</surname>
<given-names>Alexandra J.</given-names>
</name>
<xref ref-type="author-notes" rid="fn001">
<sup>*</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/359078/overview"/>
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<role content-type="https://credit.niso.org/contributor-roles/supervision/"/>
<role content-type="https://credit.niso.org/contributor-roles/project-administration/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-review-editing/"/>
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<role content-type="https://credit.niso.org/contributor-roles/conceptualization/"/>
<role content-type="https://credit.niso.org/contributor-roles/validation/"/>
</contrib>
</contrib-group>
<aff id="aff1">
<institution>Agriculture and Environmental Sciences, School of Biosciences, University of Nottingham</institution>, <addr-line>Loughborough</addr-line>,&#xa0;<country>United Kingdom</country>
</aff>
<author-notes>
<fn fn-type="edited-by">
<p>Edited by: <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/2652194/overview">Helen Wallace</ext-link>, Queensland University of Technology, Australia</p>
</fn>
<fn fn-type="edited-by">
<p>Reviewed by: <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/105035/overview">Neelendra K. Joshi</ext-link>, University of Arkansas, United States</p>
<p>
<ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/3130291/overview">Caroline Strang</ext-link>, The University of Western Ontario, Canada</p>
</fn>
<fn fn-type="corresp" id="fn001">
<p>*Correspondence: Alexandra J. Gibbs, <email xlink:href="mailto:Alexandra.gibbs@nottingham.ac.uk">Alexandra.gibbs@nottingham.ac.uk</email>
</p>
</fn>
</author-notes>
<pub-date pub-type="epub">
<day>06</day>
<month>10</month>
<year>2025</year>
</pub-date>
<pub-date pub-type="collection">
<year>2025</year>
</pub-date>
<volume>3</volume>
<elocation-id>1657493</elocation-id>
<history>
<date date-type="received">
<day>01</day>
<month>07</month>
<year>2025</year>
</date>
<date date-type="accepted">
<day>15</day>
<month>09</month>
<year>2025</year>
</date>
</history>
<permissions>
<copyright-statement>Copyright &#xa9; 2025 Dennis and Gibbs.</copyright-statement>
<copyright-year>2025</copyright-year>
<copyright-holder>Dennis and Gibbs</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/">
<p>This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</p>
</license>
</permissions>
<abstract>
<p>Neonicotinoids are systemic insecticides used in agriculture to control herbivorous pests by targeting the nervous system. However, the persistence and presence of neonicotinoids in pollen and nectar raises concerns regarding impacts on non-target organisms, particularly pollinators such as bees. Bumblebees (<italic>Bombus</italic> spp.) are essential for wild plant pollination and crop production but are vulnerable to insecticides due to their foraging behaviors and ecological traits. While commercially available forms of neonicotinoids have been banned in select countries over recent years, they are still utilized extensively in many parts of the world, with limited understanding of impacts on bumblebee physiology and behavior. To investigate neonicotinoid effects on bumblebees, we systematically reviewed studies from Scopus and Web of Science following PRISMA 2020 guidelines. A total of 52 primary studies were identified, revealing a pronounced geographic bias, with 81% of research conducted in the UK and the U.S. (54% and 27%, respectively)<italic>. Bombus terrestris, B. terrestris audax</italic>, and <italic>B. impatiens</italic> emerged as the most studied species whereas imidacloprid, thiamethoxam, and clothianidin were the most common neonicotinoid compounds tested, represented in 88% of the studies. In comparison, only a single study performed on <italic>B. ephippiatus</italic> and there are currently no published studies assessing the impact of the compounds nitenpyram or dinotefuran on bumblebee health and behavior. Behavioral alterations, particularly foraging and cognition, were the most prevalent reported effects of neonicotinoids, followed by reproductive health and physiological impacts. This review highlights the need for more geographically and taxonomically diverse research, particularly in regions still using neonicotinoids. The prevalence of sublethal effects raises concerns for colony health and pollination services, yet direct assessments of pollination efficiency remain limited. As such, critical knowledge gaps remain, particularly regarding understudied neonicotinoid compounds and bumblebee species, emphasizing the need for further research to inform sustainable agricultural practices and conservation strategies.</p>
</abstract>
<kwd-group>
<kwd>behavior</kwd>
<kwd>bumblebees (<italic>Bombus</italic> spp.)</kwd>
<kwd>colony</kwd>
<kwd>insecticides</kwd>
<kwd>neonicotinoids</kwd>
<kwd>mortality</kwd>
<kwd>physiology</kwd>
<kwd>pollinator</kwd>
</kwd-group>
<counts>
<fig-count count="6"/>
<table-count count="0"/>
<equation-count count="0"/>
<ref-count count="126"/>
<page-count count="13"/>
<word-count count="5749"/>
</counts>
<custom-meta-wrap>
<custom-meta>
<meta-name>section-in-acceptance</meta-name>
<meta-value>Bee Protection and Health</meta-value>
</custom-meta>
</custom-meta-wrap>
</article-meta>
</front>
<body>
<sec id="s1" sec-type="intro">
<label>1</label>
<title>Introduction</title>
<p>Bumblebees (<italic>Bombus</italic> spp.) are essential pollinators that contribute significantly to biodiversity and agricultural productivity (<xref ref-type="bibr" rid="B73">Parrey et&#xa0;al., 2022</xref>). They play a crucial role in pollinating various crops, including vegetables, fruits, oilseeds, legumes, fodder crops, and others that cannot be effectively pollinated by honeybees (<xref ref-type="bibr" rid="B2">Abrol et&#xa0;al., 2021</xref>; <xref ref-type="bibr" rid="B114">Wahengbam et&#xa0;al., 2019</xref>). Bumblebees have become the primary alternative to honeybees for commercial pollination in North America and Europe (<xref ref-type="bibr" rid="B35">Goulson, 2009</xref>). Their distinctive biological traits, including large body size, elongated tongue, and ability to perform buzz pollination, enhance their efficiency in pollen release and make them well-adapted to diverse environmental conditions including low temperatures and reduced light levels (<xref ref-type="bibr" rid="B8">Bie et&#xa0;al., 2025</xref>). These characteristics position bumblebees as vital pollinators, particularly in temperate regions.</p>
<p>However, bumblebee populations are experiencing alarming declines, with 24% of the 150 <italic>Bombus</italic> species assessed by the IUCN classified as threatened, and 30% of European species currently classified as of &#x201c;least concern&#x201d; are projected to lose at least 30% of ecologically suitable territory within the next 50 years (<xref ref-type="bibr" rid="B14">Cameron and Sadd, 2020</xref>; <xref ref-type="bibr" rid="B32">Ghisbain et al., 2024</xref>). This decline has been attributed to multiple stressors, including climate change, habitat loss, and pesticide exposure, interacting across spatial and temporal scales to exacerbate population losses (<xref ref-type="bibr" rid="B7">Becher et&#xa0;al., 2018</xref>). Among these threats, neonicotinoid insecticides have emerged as a critical concern due to their widespread use and documented adverse effects on pollinators.</p>
<p>The increasing global demand for food security has driven the extensive use of pesticides to enhance crop yields (<xref ref-type="bibr" rid="B75">Popp et&#xa0;al., 2013</xref>). Neonicotinoids were developed during the 1980s to overcome environmental concerns associated with older compounds such as organophosphates and combat increasing pesticide resistance among common agricultural pests (<xref ref-type="bibr" rid="B115">Werner and Hitzfeld, 2012</xref>; <xref ref-type="bibr" rid="B82">S&#xe1;nchez-Bayo et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B34">Gould et&#xa0;al., 2018</xref>). Neonicotinoids rapidly became one of the most widely used insecticides, accounting for a third of the global market within two decades (<xref ref-type="bibr" rid="B31">Ewere et&#xa0;al., 2021</xref>). Developed through nicotine-based research, neonicotinoid compounds exhibit enhanced insecticidal properties while exhibiting low toxicity to several non-target organisms such as fish, birds, and mammals (<xref ref-type="bibr" rid="B72">Pang et&#xa0;al., 2020</xref>; <xref ref-type="bibr" rid="B6">Bass and Field, 2018</xref>).</p>
<p>Neonicotinoids are classified into three primary chemical groups: N-nitroguanidines, comprising the compounds imidacloprid, thiamethoxam, clothianidin, and dinotefuran; nitromethylenes such as nitenpyram; and N-cyanoamidines including acetamiprid and thiacloprid (<xref ref-type="bibr" rid="B37">Goulson, 2013</xref>). Their versatile physicochemical properties enable diverse application approaches including foliar sprays, seed treatments, and soil drenches, making them suitable for a range of crops such as maize, sugarbeet, oilseed rape, fruits, and vegetables (<xref ref-type="bibr" rid="B70">Nauen and Jeschke, 2011</xref>; <xref ref-type="bibr" rid="B109">Van der Sluijs et&#xa0;al., 2013</xref>).</p>
<p>Neonicotinoids are characterized by their small molecular size and high water solubility, allowing them to be efficiently absorbed by plants and transported, primarily via the xylem. This systemic movement enables the compounds and their metabolites to accumulate throughout plant tissues, including within leaves, flowers, and pollen (<xref ref-type="bibr" rid="B11">Bonmatin et&#xa0;al., 2015</xref>; <xref ref-type="bibr" rid="B44">Hladik et&#xa0;al., 2018</xref>). By targeting nicotinic acetylcholine receptors (nAChRs) in the insect nervous system, neonicotinoids effectively control agricultural pests, including aphids, whiteflies, and leafhoppers (<xref ref-type="bibr" rid="B106">Tomizawa and Casida, 2005</xref>; <xref ref-type="bibr" rid="B103">Thany, 2023</xref>). While initially thought to have minimal effects on non-target organisms, evidence shows that chronic exposure impairs pollinator foraging, reproduction, and colony health, and they are often associated with high bee toxicities (<xref ref-type="bibr" rid="B45">Hopwood et&#xa0;al., 2012</xref>; <xref ref-type="bibr" rid="B100">Stokstad, 2013</xref>; <xref ref-type="bibr" rid="B6">Bass and Field, 2018</xref>; <xref ref-type="bibr" rid="B48">Jeschke et&#xa0;al., 2011</xref>; <xref ref-type="bibr" rid="B5">Basley, 2019</xref>; <xref ref-type="bibr" rid="B87">Schulz et&#xa0;al., 2021</xref>).</p>
<p>The global decline in bee populations has raised significant concerns due to their vital role in pollination and the potential consequences for ecosystems and agriculture. As part of this, increasing research has highlighted the harmful effects of neonicotinoids, particularly on bee foraging, behavior, reproduction, and pollination services (<xref ref-type="bibr" rid="B47">Ihara and Matsuda, 2018</xref>; <xref ref-type="bibr" rid="B76">Pyke, 2022</xref>). Consequently, in 2018, the European Union (EU) banned all outdoor uses of clothianidin, imidacloprid, and thiamethoxam (although some exemptions were permitted up until 2024; <xref ref-type="bibr" rid="B22">Dentzman et&#xa0;al., 2025</xref>), and in 2020, it withheld renewal of thiacloprid&#x2019;s approval due to endocrine disruption risks. The U.S. Environmental Protection Agency (EPA) cancelled 12 neonicotinoid products in 2019 with many state-level restrictions in place since 2023 (<xref ref-type="bibr" rid="B107">U.S. EPA, 2019</xref>; <xref ref-type="bibr" rid="B69">National Caucus of Environmental Legislators, 2023</xref>) and Canada&#x2019;s Pest Management Regulatory Agency (PMRA) proposed a series of risk mitigation measures and restrictions on use, with reevaluation of measures expected to be published in 2026 (<xref ref-type="bibr" rid="B42">Health Canada, 2020</xref>; <xref ref-type="bibr" rid="B43">Health Canada, 2025</xref>; <xref ref-type="bibr" rid="B22">Dentzman et&#xa0;al., 2025</xref>). In addition, the joint FAO/WHO Meeting on Pesticide Residues introduced intake limits to reduce health risks (<xref ref-type="bibr" rid="B105">Thompson et&#xa0;al., 2020</xref>). Despite these regulatory measures, neonicotinoids remain widely used in regions lacking pesticide legislation, with clothianidin, imidacloprid, and thiamethoxam still dominating the global market (<xref ref-type="bibr" rid="B51">Klingelh&#xf6;fer et&#xa0;al., 2022</xref>).</p>
<p>Despite growing evidence for the negative impact of neonicotinoid use, knowledge gaps remain, particularly for less studied taxa such as bumblebees (<xref ref-type="bibr" rid="B25">Dirilgen et&#xa0;al., 2023</xref>). However, given the ecological importance of bumblebees and the increasing reliance on pollination services for food production, understanding the extent of the impact of neonicotinoids is essential for informing sustainable agricultural practices and conservation efforts. Therefore, this systematic review aims to critically evaluate existing research on the effects of neonicotinoids on bumble bees, synthesizing both laboratory and field findings. We aimed to 1) identify which compound is the most studied, 2) identify the taxonomic spread of studies using neonicotinoids, and 3) assess the main reported impacts on bumblebee health and physiology. By assessing the broader ecological implications and identifying research gaps, this review will contribute to ongoing discussions on pollinator protection, regulatory measures, and future pesticide use.</p>
</sec>
<sec id="s2">
<label>2</label>
<title>Methods</title>
<p>A systematic review was conducted following the PRISMA 2020 guidelines. A comprehensive literature search was performed using two academic databases: Scopus and Web of Science, with the most recent search performed in June 2025. To ensure all relevant literature was captured, the following search string and Boolean operators were applied to both databases: (&#x201c;Neonic*&#x201d; OR &#x201c;Neonicotinoid&#x201d; OR &#x201c;Neonicotinoids&#x201d; OR &#x201c;Imidacloprid&#x201d; OR &#x201c;Clothianidin&#x201d; OR &#x201c;Thiamethoxam&#x201d; OR &#x201c;Acetamiprid&#x201d; OR &#x201c;Thiacloprid&#x201d; OR &#x201c;Nitenpyram&#x201d; OR &#x201c;Dinotefuran&#x201d;) AND (&#x201c;Bumblebee*&#x201d; OR &#x201c;Bumble bee*&#x201d; OR &#x201c;<italic>Bombus</italic>&#x201d;). The search was restricted to peer-reviewed, open-access articles within the last 20 years to ensure the review captured recent research. Only English papers were considered.</p>
<p>Following the database search, titles and abstracts were screened independently against the inclusion and exclusion criteria. Briefly, studies were included if they focused on bumblebees, examined neonicotinoid effects, and assessed behavioral, physiological, reproductive, survival, or pollination impacts under field-realistic or controlled conditions. Studies on other pollinators, non-neonicotinoid pesticides, or multiple interacting stressors were excluded.</p>
<p>Duplicates were removed, and full texts of potentially relevant papers were reviewed to confirm eligibility (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). For each included study, the publication date was noted (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2</bold>
</xref>), and data were extracted on neonicotinoid compound (<xref ref-type="fig" rid="f3">
<bold>Figure&#xa0;3</bold>
</xref>), study location (<xref ref-type="fig" rid="f4">
<bold>Figure&#xa0;4</bold>
</xref>), bumblebee species (<xref ref-type="fig" rid="f5">
<bold>Figure&#xa0;5</bold>
</xref>), and measured impact on bumblebee health or physiology, classed into behavioral, physiological, reproductive, survival, and mortality or pollination efficiency-related effects (<xref ref-type="fig" rid="f6">
<bold>Figure&#xa0;6</bold>
</xref>).</p>
<fig id="f1" position="float">
<label>Figure&#xa0;1</label>
<caption>
<p>Flowchart of studies selected for the systematic analysis of the impact of neonicotinoids on bumblebees adapted from the PRISMA 2020 guidelines.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="frbee-03-1657493-g001.tif">
<alt-text content-type="machine-generated">Flowchart depicting the study selection process. Initial identification from Scopus and Web of Science yielded 6,072 records, reduced to 2,827 after removing 3,245 ineligible records. Post-screening, 2,703 records were excluded based on criteria, leaving 124 for retrieval. After removing 48 duplicates, 77 records were assessed for eligibility. Additional exclusions due to various reasons resulted in 52 studies included in the review, plus 2 forward citations.</alt-text>
</graphic>
</fig>
<fig id="f2" position="float">
<label>Figure&#xa0;2</label>
<caption>
<p>Development of research on the effect of neonicotinoids on bumblebees over time according to compound studied.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="frbee-03-1657493-g002.tif">
<alt-text content-type="machine-generated">Stacked bar graph showing the number of studies published per year between 2012 and 2025, split by pesticide compound studied. Each bar consists of sections: orange for Imidacloprid, blue for Thiamethoxam, green for Clothianidin, yellow for Acetamiprid, red for Thiacloprid, gray for multiple, and black for not stated. The number of studies ranges from zero to seven per year.</alt-text>
</graphic>
</fig>
<fig id="f3" position="float">
<label>Figure&#xa0;3</label>
<caption>
<p>Frequency distribution of studied neonicotinoid compounds across 52 systematically identified studies with percentage of total studies indicated.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="frbee-03-1657493-g003.tif">
<alt-text content-type="machine-generated">Bar chart displaying the number of studies for the effect on bumblebees for different neoinicitoind compounds. Imidacloprid leads with 54 percent, followed by Thiamethoxam at 23 percent, Clothianidin at 21 percent, Acetamiprid at 10 percent, and Thiacloprid at 6 percent.</alt-text>
</graphic>
</fig>
<fig id="f4" position="float">
<label>Figure&#xa0;4</label>
<caption>
<p>Geographical distribution of research on neonicotinoid impacts on bumblebees. Colors indicate countries ranging from 1 study (yellow) to &gt;28 studies (pink).</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="frbee-03-1657493-g004.tif">
<alt-text content-type="machine-generated">World map highlighting the number of studies conducted by country, using a gradient from yellow to red. The United States, Canada, and China show a significant number of studies, while the United Kingdom highlights the highest density. Other countries in Europe, Asia, and North America display varying levels of study frequencies. The color legend indicates quantities ranging from five to over twenty-five studies.</alt-text>
</graphic>
</fig>
<fig id="f5" position="float">
<label>Figure&#xa0;5</label>
<caption>
<p>Frequency distribution of bumblebee taxa in the research literature regarding the impact of neonicotinoids on health and physiology across 51 systematically identified studies (one published study does not report on species used).</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="frbee-03-1657493-g005.tif">
<alt-text content-type="machine-generated">Bar chart showing the number of studies on different bumblebee species. Bombus terrestris has the highest count near 19, followed by Bombus impatiens at 14. Bombus terrestris audax is next with 5, and Bombus ephippiatus is lowest with with 1 study.</alt-text>
</graphic>
</fig>
<fig id="f6" position="float">
<label>Figure&#xa0;6</label>
<caption>
<p>Frequency distribution of the observed neonicotinoid effects on bumblebee health and physiology across 52 systematically identified studies, with the percentage of total studies indicated.</p>
</caption>
<graphic mimetype="image" mime-subtype="tiff" xlink:href="frbee-03-1657493-g006.tif">
<alt-text content-type="machine-generated">Bar chart depicting the number of studies by reported impact category. Categories include Behavioral (52%), Reproductive (40%), Physiological (34%), Survival and Mortality (27%), and Pollination Efficiency (11%). The y-axis shows the number of studies from 0 to 30.</alt-text>
</graphic>
</fig>
</sec>
<sec id="s3" sec-type="results">
<label>3</label>
<title>Results</title>
<p>A total of 52 original research publications matched our criteria (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>). Full references for all publications and data for each primary research publication are presented in <xref ref-type="supplementary-material" rid="SM1">
<bold>Supplementary Table S1</bold>
</xref>. Relevant literature was published from 2012 onward, with approximately half of the studies published within the last 6 years, demonstrating a rapid expansion of the research field (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2</bold>
</xref>). The highest number of studies was published in 2021 (n = 6).</p>
<p>Imidacloprid was the most commonly studied compound (included in 54% of studies, n = 28), followed by thiamethoxam (23%, n = 12) and clothianidin (19%, n = 11; <xref ref-type="fig" rid="f3">
<bold>Figure&#xa0;3</bold>
</xref>). Acetamiprid and thiacloprid were less frequently studied, accounting for 10% (n = 5) and 6% (n = 3).</p>
<p>Primary research studies were conducted in 10 countries. However, 81% were performed in just two countries: United Kingdom (n = 28) and the United States (n = 14; <xref ref-type="fig" rid="f4">
<bold>Figure&#xa0;4</bold>
</xref>). Two studies were conducted in Switzerland and Norway, whereas single studies were conducted in the Netherlands, Belgium, Finland, Israel, Mexico, China, and Canada.</p>
<p>Studies included in this review represented four bumblebee taxa (<xref ref-type="fig" rid="f5">
<bold>Figure&#xa0;5</bold>
</xref>). <italic>Bombus terrestris</italic> was the most frequently studied (n = 20), followed by its subspecies <italic>Bombus terrestris audax</italic> (n = 16). <italic>Bombus impatiens</italic> was featured in 15 studies, whereas <italic>Bombus ephippiatus</italic> was notably underrepresented, with only one published study.</p>
<p>Behavioral effects were the most frequently reported impact of neonicotinoids on bumblebees, comprising 52% of studies (n = 27), followed by reproductive (40%, n = 21) and physiological effects (35%, n = 18; <xref ref-type="fig" rid="f6">
<bold>Figure&#xa0;6</bold>
</xref>). Survival and mortality outcomes were documented in 27% of studies (n = 14), whereas pollination efficiency and ecosystem impacts were the least observed, reported in 11% of studies (n = 6).</p>
</sec>
<sec id="s4" sec-type="discussion">
<label>4</label>
<title>Discussion</title>
<p>With pollinator declines globally recognized, understanding the specific effects of neurotoxic pesticides on bumblebee health and behavior is essential. This systematic review identified 52 studies investigating the impacts of neonicotinoids on bumblebees (<xref ref-type="fig" rid="f1">
<bold>Figure&#xa0;1</bold>
</xref>).</p>
<sec id="s4_1">
<label>4.1</label>
<title>Neonicotinoid bans and ecological events drive research on bumblebee impacts</title>
<p>The increasing publication trend from 2012 to 2021 (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2</bold>
</xref>) likely reflects growing scientific scrutiny around the use of neonicotinoids. The EU applied a partial ban of neonicotinoids in 2013, followed by the complete ban of three compounds in outdoor field crops in 2018 (<xref ref-type="bibr" rid="B88">Sgolastra et&#xa0;al., 2020</xref>; <xref ref-type="bibr" rid="B22">Dentzman et&#xa0;al., 2025</xref>). During 2019-2023, emergency pesticide exemptions were made to allow the use of imidacloprid and thiamethoxam in sugar beet, but this decision was reversed in 2023 following appeals (<xref ref-type="bibr" rid="B30">Epstein et&#xa0;al., 2022</xref>). Exemptions for neonicotinoid use in the United Kingdom (which left the European Union in 2021) were feasible up to 2024 (<xref ref-type="bibr" rid="B18">Coe et&#xa0;al., 2024</xref>; <xref ref-type="bibr" rid="B21">DEFRA, 2025</xref>). Currently, the use of cyanoamidine neonicotinoids has not been restricted in the EU, and use of acetamiprid is approved until February 2033, with approval of sulfoxaflor and flupyradifurone, up for renewal in 2025 (<xref ref-type="bibr" rid="B22">Dentzman et&#xa0;al., 2025</xref> and references within).</p>
<p>While slower to implement restrictions on the use of neonicotinoids than the European Union, state-level bans have been implemented (<xref ref-type="bibr" rid="B107">U.S. EPA, 2019</xref>; <xref ref-type="bibr" rid="B22">Dentzman et&#xa0;al., 2025</xref>). As of 2023, at least 20 states have imposed some form of restriction to protect pollinators (<xref ref-type="bibr" rid="B69">National Caucus of Environmental Regulators, 2023</xref>).</p>
<p>The peak in publications in 2021 could have been influenced by the resurgence of virus yellows in sugar beet in 2020 following the EU ban on neonicotinoid seed treatments (<xref ref-type="bibr" rid="B24">Dewar and Qi, 2021</xref>), prompting further investigation into the broader consequences of these regulations. Publication delays (<xref ref-type="bibr" rid="B9">Bj&#xf6;rk and Solomon, 2013</xref>) also suggest that research following the bans likely contributed to the later publication surge.</p>
</sec>
<sec id="s4_2">
<label>4.2</label>
<title>Neonicotinoid compounds studied reflect commercial use and toxicity profiles: imidacloprid, thiamethoxam, and clothianidin</title>
<p>Imidacloprid, thiamethoxam, and clothianidin were the most frequently studied neonicotinoids (<xref ref-type="fig" rid="f2">
<bold>Figures&#xa0;2</bold>
</xref>, <xref ref-type="fig" rid="f3">
<bold>3</bold>
</xref>). This may reflect the greater risk of nitroguanidine neonicotinoids on pollinators relative to the lower threat cyanoamidine group (<xref ref-type="bibr" rid="B12">Buszewski et&#xa0;al., 2019</xref>). An alternative explanation for the prevalence of studies on these compounds has been proposed by <xref ref-type="bibr" rid="B57">Lundin et&#xa0;al. (2015)</xref>, who identified a positive correlation between the number of studies on each neonicotinoid and its global sales, indicating that compounds with higher commercial usage tend to be more extensively researched. This aligns with findings from <xref ref-type="bibr" rid="B104">Thompson (2020)</xref> who identified that in 2012, imidacloprid, thiamethoxam, and clothianidin accounted for 85% of all neonicotinoids sold. Of the three compounds, imidacloprid is the most widely used neonicotinoid, with registered applications on over 140 crops across 120 countries (<xref ref-type="bibr" rid="B89">Sheets, 2010</xref>; <xref ref-type="bibr" rid="B48">Jeschke et&#xa0;al., 2011</xref>), explaining high research attention. An additional factor which may contribute to the prevalence of imidacloprid prevalence in research may be linked to their strong binding affinity for nAChRs, which influences their toxicological effects and associated risks to bees (<xref ref-type="bibr" rid="B63">Moffat et&#xa0;al., 2016</xref>).</p>
<p>In comparison, acetamiprid has been studied less extensively due to its classification as a lower-risk neonicotinoid. Regulatory assessments have found it to pose minimal toxicity to bees at field-realistic concentrations, leading to its continued unrestricted use in the EU since 2021 (<xref ref-type="bibr" rid="B46">House of Commons, 2022</xref>). This coincides with the two published studies on acetamiprid in 2020 and 2022 (<xref ref-type="fig" rid="f2">
<bold>Figure&#xa0;2</bold>
</xref>). While high doses can cause sublethal effects, its lower overall risk has contributed to reduced research focus (<xref ref-type="bibr" rid="B111">Varga-Szilay and T&#xf3;th, 2022</xref>).</p>
<p>Similarly, thiacloprid was among the least studied neonicotinoids, likely due to its comparatively lower acute toxicity to bees and its classification as relatively &#x201c;bee-safe&#x201d; (<xref ref-type="bibr" rid="B13">Cabezas and Farin&#xf3;s, 2022</xref>; <xref ref-type="bibr" rid="B29">Ellis et&#xa0;al., 2017</xref>). Although residue concentrations can be high, the extended time to reach lethal doses reduces immediate risk (<xref ref-type="bibr" rid="B13">Cabezas and Farin&#xf3;s, 2022</xref>). Despite this, approval for agricultural use of thiacloprid was discontinued in the EU in 2021, with limited ongoing use in greenhouses (<xref ref-type="bibr" rid="B4">Bai et&#xa0;al., 2025</xref>).</p>
<p>Dinotefuran and nitenpyram were absent from the studies identified in this review, likely due to their less prevalent use in agriculture (<xref ref-type="bibr" rid="B15">Camp and Lehmann, 2021</xref>). While dinotefuran has been implicated in bee mortality events, research on its sublethal effects remains limited (<xref ref-type="bibr" rid="B41">Hatfield et&#xa0;al., 2021</xref>). In contrast, nitenpyram is primarily used as a veterinary flea treatment; however, there have been reported adverse effects on non-target organisms (<xref ref-type="bibr" rid="B79">Rust et&#xa0;al., 2003</xref>; <xref ref-type="bibr" rid="B126">Zhu et&#xa0;al., 2020</xref>).</p>
<p>
<xref ref-type="bibr" rid="B45">Hopwood et&#xa0;al. (2012)</xref> show that imidacloprid and clothianidin exhibit high acute toxicity to bumblebees. Although precise LD50 (lethal dose 50%: a determined dose, expected to cause mortality in 50% of treated subjects; <xref ref-type="bibr" rid="B67">Myers, 2017</xref>) values have not been established, research indicates that both neonicotinoids are highly harmful upon acute contact exposure. Additionally, imidacloprid is highly toxic when administered orally. Comparatively, clothianidin has slightly higher toxicity than imidacloprid through contact exposure. <xref ref-type="bibr" rid="B45">Hopwood et&#xa0;al. (2012)</xref> suggest that the acute toxicity of other neonicotinoids, including acetamiprid, dinotefuran, thiacloprid, and thiamethoxam, remain untested.</p>
<p>The wide range in neonicotinoid toxicities for both target and non-target organisms poses challenges for the regulatory risk assessment conducted during pesticide authorization. In particular, the lesser known impacts of the cyanoamidine group limits our understanding of the wider scale impact of neonicotinoid use (<xref ref-type="bibr" rid="B40">Grout et&#xa0;al., 2020</xref>; <xref ref-type="bibr" rid="B22">Dentzman et&#xa0;al., 2025</xref>). As such, risk assessments are usually based upon limited standard test species (<xref ref-type="bibr" rid="B108">U.S. EPA, 2021</xref>; <xref ref-type="bibr" rid="B28">EFSA, 2013</xref>). As such, regulatory threshold levels (RTLs), which determine the acute and chronic thresholds for unacceptable ecological effects, may underestimate actual risk to diverse taxa.</p>
</sec>
<sec id="s4_3">
<label>4.3</label>
<title>The geographical distribution of publications shows a bias toward the UK and U.S.</title>
<p>The geographical distribution of research investigating the effects of neonicotinoids on bumblebees exhibits a notable bias, with most studies concentrated in the United Kingdom and the United States (<xref ref-type="fig" rid="f4">
<bold>Figure&#xa0;4</bold>
</xref>). This may reflect capacity and interest in bee-related research, their prominence as major agricultural producers, and also policy requirements for the application of insecticides, which has driven scientific inquiry into the ecological impacts (<xref ref-type="bibr" rid="B26">Donley, 2019</xref>). In the U.S. alone, over 1,000 neonicotinoid-containing products are registered with the EPA and remain available on the market (<xref ref-type="bibr" rid="B104">Thompson, 2020</xref>). Meanwhile, the EU&#x2019;s ban may have intensified research efforts to assess the ecological risks posed by these chemicals.</p>
<p>Bumblebees are particularly abundant in temperate regions, where their adaptation to cooler climates makes them key pollinators in these ecosystems (<xref ref-type="bibr" rid="B35">Goulson, 2009</xref>). The regional focus is also consistent with bumblebee population trends, as species declines have been widely documented across Europe and North America (<xref ref-type="bibr" rid="B120">Williams and Osborne, 2009</xref>). The UK has lost three of its 27 native bumblebee species, with seven more designated as priority species due to significant declines (<xref ref-type="bibr" rid="B77">Reid et&#xa0;al., 2020</xref>). These population declines further incentivized research efforts in these regions, as the potential consequences of pollinator losses for ecosystems and food security have become an increasing concern.</p>
<p>Despite extensive research in Europe and North America, studies remain limited in other regions. East Asia, for example, hosts the richest bumblebee diversity globally, with over 50% of known species and approximately 10% of the world&#x2019;s endemic species concentrated in countries such as China, Mongolia, Japan, and parts of Russia and India (<xref ref-type="bibr" rid="B68">Naeem et&#xa0;al., 2019</xref>). Similarly, South America supports diverse bumblebee populations (<xref ref-type="bibr" rid="B53">Krechemer et&#xa0;al., 2020</xref>). However, as noted by <xref ref-type="bibr" rid="B25">Dirilgen et&#xa0;al. (2023)</xref>, stingless bees (Apidae: Apinae: Meliponini) are more commonly used for commercial pollination in South America which could explain the lack of research on bumblebees.</p>
<p>Research in these regions is crucial, particularly as warmer climates typically support higher ectotherm populations, including insect pests and their associated pathogens (<xref ref-type="bibr" rid="B17">Clarke and P&#xf6;rtner, 2010</xref>; <xref ref-type="bibr" rid="B91">Simon and Amarasekare, 2024</xref>). Increased temperatures accelerate insect metabolic rates, leading to heightened food consumption and population growth, escalating pest pressure and reliance on insecticides (<xref ref-type="bibr" rid="B23">Deutsch et&#xa0;al., 2018</xref>; <xref ref-type="bibr" rid="B85">Schneider et&#xa0;al., 2022</xref>). Additionally, the economic challenges faced by warmer, less developed countries (<xref ref-type="bibr" rid="B110">Van de Vliert et&#xa0;al., 2000</xref>) may increase their vulnerability to agricultural losses and subsequent reliance on broad-spectrum insecticides.</p>
<p>Globally, the continued widespread use of the three major neonicotinoids is facilitated by the absence of pesticide legislation in approximately 35% of countries (<xref ref-type="bibr" rid="B51">Klingelh&#xf6;fer et&#xa0;al., 2022</xref>). This may, in part, be a result of a lack of national-level reporting of pesticide use in many countries. For example, <xref ref-type="bibr" rid="B102">Tai et&#xa0;al. (2025)</xref> suggested that a lack of centralized collection for pesticide data in New Zealand may contribute to masking the true scale and frequency of neonicotinoid use and that current legislation is often restricted to a single taxonomic group. In the case of New Zealand, legislation is restricted to the impact of pesticide use on honeybees only, despite the presence of four bumblebee species and many ground-dwelling native bees (<xref ref-type="bibr" rid="B102">Tai et&#xa0;al., 2025</xref>). Lack of regulation or centralized collection has also been reported for many parts of Africa and Asia (<xref ref-type="bibr" rid="B118">WHO, 2018</xref>).</p>
</sec>
<sec id="s4_4">
<label>4.4</label>
<title>The bumblebee species investigated indicate a taxonomic bias toward commercially significant species: <italic>Bombus terrestris</italic> and <italic>Bombus impatiens</italic>
</title>
<p>The predominant focus on <italic>Bombus terrestris, Bombus terrestris audax</italic>, and <italic>Bombus impatiens</italic> in neonicotinoid research (<xref ref-type="fig" rid="f5">
<bold>Figure&#xa0;5</bold>
</xref>) likely reflects their ecological distribution and commercial significance. <italic>Bombus terrestris</italic> has a broad natural range across Europe, North Africa, and the Near East (<xref ref-type="bibr" rid="B119">Widmer et&#xa0;al., 1998</xref>). Since 1988, <italic>B. terrestris</italic> has been commercially reared for greenhouse crop pollination in Europe, driving research interest due to their ecological and economic importance (<xref ref-type="bibr" rid="B33">Gosterit and Baskar, 2016</xref>). Their ability to establish large colonies and adapt to artificial conditions makes them a practical species for commercial and experimental purposes (<xref ref-type="bibr" rid="B112">Velthuis and van Doorn, 2004</xref>). Likewise, <italic>B. terrestris audax</italic>, a subspecies endemic to Britain and Ireland, is heavily studied in the UK, aligning with the country&#x2019;s prominence in neonicotinoid research (<xref ref-type="bibr" rid="B36">Goulson, 2010</xref>).</p>
<p>In North America, <italic>Bombus impatiens</italic> is the primary commercially significant species (<xref ref-type="bibr" rid="B121">Winter et&#xa0;al., 2006</xref>). Its widespread use in agricultural pollination likely explains its frequent inclusion in neonicotinoid studies. Conversely, <italic>Bombus ephippiatus</italic> is a Mesoamerican species with limited representation in research, likely due to its restricted distribution (<xref ref-type="bibr" rid="B27">Duennes et&#xa0;al., 2012</xref>). However, considering there are over 260 known species worldwide, with native species in all biogeographic regions except for Australia, New Zealand, and sub-Saharan Africa (<xref ref-type="bibr" rid="B14">Cameron and Sadd, 2020</xref>); this taxonomic bias indicates critical gaps in our understanding of neonicotinoids on bumblebee health and behavior.</p>
<p>Understanding species-level differences in ecotoxicity is important for both scientific understanding and regulation (<xref ref-type="bibr" rid="B49">J&#xfc;tte et&#xa0;al., 2023</xref>). While <italic>B. terrestris</italic> and <italic>B. impatiens</italic> are commercially available, easy to rear and convenient for experimentation, they may not be representative of wild species with smaller populations or different ecologies. For example, <italic>B. terrestris</italic> has large colonies containing hundreds of workers, which may help buffer against loss. In comparison, <italic>B. pseudobaicalensis</italic> and <italic>B. schrencki</italic> have smaller colonies (<xref ref-type="bibr" rid="B20">Cueva Del Castillo et&#xa0;al., 2015</xref>) and therefore may be more vulnerable, since losing a few foragers can mean a significant impact on resource intake.</p>    <p>Larger-bodied bumblebees often travel further when foraging (<xref ref-type="bibr" rid="B39">Greenleaf et&#xa0;al., 2007</xref>), so any disruptions to navigation or endurance from neonicotinoids may have a greater impact on these species. In addition, it is possible that there will be a difference based on whether a species is a generalist or a specialist when it comes to pollination, with a narrower floral niche which includes impacted plants equating to increased risk of exposure (<xref ref-type="bibr" rid="B49">J&#xfc;tte et&#xa0;al., 2023</xref>). Differences may also relate to the timing of colony activity, with early emerging species more likely to be exposed to high neonicotinoid residues during spring sowing.</p>
<p>It is also possible that species may metabolize insecticides at different rates, making some more resistant than others (<xref ref-type="bibr" rid="B125">Zarevcka, 2013</xref>; <xref ref-type="bibr" rid="B58">Manjon et&#xa0;al., 2018</xref>). This reflects the findings of <xref ref-type="bibr" rid="B49">J&#xfc;tte et&#xa0;al. (2023)</xref> who found that field-realistic concentrations of a pyrethroid insecticide had differential effects between multiple bee species which they attributed to individual bee weight combined with additional ecological, phylogenetic, or toxicogenomic parameters.</p>
<p>As bumblebees fill different pollination niches, if neonicotinoids selectively impact some species more than others, plant&#x2013;pollinator networks can become unbalanced, potentially resulting in some plants losing their most effective pollinators. Therefore, risk assessments that rely on a single species underestimate the variability of effects and including multiple species ensures that pesticide approvals account for the most vulnerable pollinators (<xref ref-type="bibr" rid="B49">J&#xfc;tte et&#xa0;al., 2023</xref>).</p>
</sec>
<sec id="s4_5">
<label>4.5</label>
<title>Observed effects of neonicotinoids disrupt bumblebee behavior, reproductive success, and ecosystem functioning</title>
<p>Neonicotinoid exposure has been shown to disrupt key bumblebee behaviors (<xref ref-type="fig" rid="f6">
<bold>Figure&#xa0;6</bold>
</xref>). Of our identified studies, foraging behavior was the most frequently reported change with
affected bees exhibiting reduced flower visitation rates (<xref ref-type="bibr" rid="B95">Stanley
et&#xa0;al., 2015</xref>), altered flower preferences (<xref ref-type="bibr" rid="B93">Siviter et&#xa0;al., 2021</xref>), and decreased foraging motivation (<xref ref-type="bibr" rid="B54">L&#xe4;ms&#xe4; et&#xa0;al., 2018</xref>; <xref ref-type="bibr" rid="B66">Muth and Leonard, 2019</xref>). Chronic exposure can also impair foraging efficiency by reducing flight endurance, increasing exhaustion rates (<xref ref-type="bibr" rid="B84">Sargent et al., 2021</xref>), and affecting homing ability (<xref ref-type="bibr" rid="B97">Stanley et&#xa0;al., 2016</xref>; <xref ref-type="bibr" rid="B50">Kenna et&#xa0;al., 2019</xref>). Such disruptions can cascade through the colony, reducing resource intake and impairing colony growth.</p>    <p>Learning and memory deficits further exacerbate changes in behavior, with exposed bumblebees demonstrating impaired associative learning, olfactory conditioning (<xref ref-type="bibr" rid="B95">Stanley et&#xa0;al., 2015</xref>; <xref ref-type="bibr" rid="B65">Muth et&#xa0;al., 2019</xref>), and spatial memory (<xref ref-type="bibr" rid="B81">Samuelson et&#xa0;al., 2016</xref>). Since bumblebees rely on learned foraging routes to locate and return to high-quality floral resources, these cognitive impairments likely contribute to colony-wide reductions in food intake. Additionally, exposure reduces responsiveness to food rewards (<xref ref-type="bibr" rid="B94">Smith et&#xa0;al., 2020</xref>), decreases sucrose and nectar consumption (<xref ref-type="bibr" rid="B74">Paus-Knudsen et&#xa0;al., 2023</xref>; <xref ref-type="bibr" rid="B56">Laycock et&#xa0;al., 2012</xref>, <xref ref-type="bibr" rid="B55">Laycock et&#xa0;al., 2014</xref>), and diminishes food storage within the colony (<xref ref-type="bibr" rid="B86">Scholer and Krischik, 2014</xref>). Disruptions in essential social behaviors, such as nest thermoregulation (<xref ref-type="bibr" rid="B19">Crall et&#xa0;al,  2018</xref>) and brood care (<xref ref-type="bibr" rid="B16">Chole et&#xa0;al., 2022</xref>), further weaken colony cohesion, reducing reproductive success and survival.</p>
<p>Reproductive effects were also commonly reported, with exposed colonies producing fewer new queens, limiting their ability to establish future generations (<xref ref-type="bibr" rid="B116">Whitehorn et&#xa0;al., 2012</xref>; <xref ref-type="bibr" rid="B38">Goulson, 2015</xref>). Brood production and overall colony growth were often impaired, with fewer workers and reproductive individuals emerging (<xref ref-type="bibr" rid="B16">Chole et&#xa0;al., 2022</xref>; <xref ref-type="bibr" rid="B60">Mart&#xed;nez de Castro Dubernard et&#xa0;al., 2022</xref>; <xref ref-type="bibr" rid="B122">Wintermantel et&#xa0;al., 2018</xref>). Delayed colony development, including slower nest initiation and egg laying (<xref ref-type="bibr" rid="B124">Wu-Smart and Spivak, 2018</xref>), further compromises population stability. Male reproduction was also affected, with reduced drone weight, disrupted production timing, and impaired sperm viability limiting mating success (<xref ref-type="bibr" rid="B101">Straub et&#xa0;al., 2022</xref>; <xref ref-type="bibr" rid="B62">Minnameyer et&#xa0;al., 2021</xref>). These impairments align with previous research demonstrating neonicotinoid-induced colony decline but extend existing knowledge by emphasizing the vulnerability of reproductive individuals.</p>
<p>Physiologically, neonicotinoids disrupt neurophysiology by interfering with nAChRs, impairing neural development (<xref ref-type="bibr" rid="B59">Mart&#xed;n-Bl&#xe1;zquez et&#xa0;al., 2023</xref>), and altering brain ion transport-related gene expression (<xref ref-type="bibr" rid="B123">Witwicka et&#xa0;al., 2025</xref>). Increased acetylcholinesterase expression (<xref ref-type="bibr" rid="B80">Samson-Robert et&#xa0;al., 2015</xref>) suggests impaired neurotransmission, potentially explaining deficits in foraging efficiency and learning ability. Metabolic effects include increased energy demands and reduced detoxification capacity, with exposure elevating oxygen consumption and metabolic activity in flight muscles and the brain (<xref ref-type="bibr" rid="B84">Sargent et&#xa0;al., 2021</xref>). Together, these changes may lead to hyperactivity and exhaustion. Reduced detoxification efficiency at higher neonicotinoid concentrations (<xref ref-type="bibr" rid="B1">Aar&#xf8;nes et&#xa0;al., 2021</xref>) suggests a threshold where bees struggle to process toxins, increasing overall physiological stress. In addition, exposure also alters nectar-related immune responses (<xref ref-type="bibr" rid="B78">Richman et&#xa0;al., 2022</xref>) and upregulates antimicrobial peptide gene expression (<xref ref-type="bibr" rid="B90">Simmons and Angelini, 2017</xref>), which could lead to immune overactivation and long-term susceptibility to pathogens.</p>
<p>Survival and mortality effects have been documented, with exposed workers experiencing reduced lifespan and colonies facing premature failure (<xref ref-type="bibr" rid="B55">Laycock et&#xa0;al., 2014</xref>; <xref ref-type="bibr" rid="B29">Ellis et&#xa0;al., 2017</xref>). Even sublethal doses shorten lifespan by altering nectar chemistry (<xref ref-type="bibr" rid="B78">Richman et&#xa0;al., 2022</xref>), which may accelerate colony collapse. Although studied less, widespread bee mortality in certain regions has been associated with large-scale neonicotinoid use (<xref ref-type="bibr" rid="B51">Klingelh&#xf6;fer et&#xa0;al., 2022</xref>).</p>
<p>Direct measures of pollination efficiency and ecosystem impacts were less observed, which is likely a result of few studies being performed in the field relative to laboratory conditions (five out of 52; <xref ref-type="supplementary-material" rid="SM1">
<bold>Supplementary Table S1</bold>
</xref>; <xref ref-type="bibr" rid="B49">J&#xfc;tte et&#xa0;al., 2023</xref>). Other potential reasons for reduced focus in this research area could be the complexity of experimental design required; timeframes needed for study; and potential conflicting factors (beyond neonicotinoid application), which may limit the ability to make a causal link to insecticide use. Additionally, funding and policy requirements have driven focus on establishing whether neonicotinoids directly harm bees, as opposed to longer-term ecosystem-scale studies which are more costly and uncertain.</p>
<p>In contrast, more studies reported behavioral impairments, reduced reproductive success, and increased mortality in bumblebees, traits which are inherently linked to pollination performance and ecosystem functioning. Behavioral effects such as reduced and altered foraging can lead to fewer flower visits and diminished pollen collection (<xref ref-type="bibr" rid="B95">Stanley et&#xa0;al., 2015</xref>; <xref ref-type="bibr" rid="B54">L&#xe4;ms&#xe4; et&#xa0;al., 2018</xref>; <xref ref-type="bibr" rid="B52">Kolano et&#xa0;al., 2021</xref>). Compromised buzz pollination efficiency, with shorter buzzing duration, results in lower pollen deposition and reduced crop yield (<xref ref-type="bibr" rid="B117">Whitehorn et&#xa0;al., 2017</xref>; <xref ref-type="bibr" rid="B95">Stanley et&#xa0;al., 2015</xref>). Furthermore, smaller or poorly structured colonies result in fewer foragers, limiting floral visitation and pollen transfer (<xref ref-type="bibr" rid="B61">McGrady et&#xa0;al., 2021</xref>). This reduced reproductive output also compromises the establishment of future colonies, contributing to long-term pollinator population declines. Similarly, increased mortality, particularly among foragers, reduces the duration and scale of pollination activity within the colony, further weakening pollination services. At a broader scale, exposure disrupts plant&#x2013;pollinator networks, with altered interactions between bumblebees and wild plants, potentially affecting plant reproductive success and overall ecosystem stability (<xref ref-type="bibr" rid="B96">Stanley and Raine, 2016</xref>; <xref ref-type="bibr" rid="B3">Arce et&#xa0;al., 2017</xref>).</p>
<p>These findings reinforce concerns about the widespread ecological risks of neonicotinoids while highlighting key gaps in current knowledge. The predominance of behavioral effects within the literature emphasizes that the sublethal impacts on foraging, learning, and social behaviors may be more consequential than previously acknowledged. While reproductive and physiological effects were also substantial, survival and pollination impacts were underreported. This suggests that subtle, chronic impairments in behavior and reproduction could undermine colony health before direct mortality is observed. Given that pollination efficiency and ecosystem impacts were the least observed, further research is needed to explore how other disruptions translate into broader ecological consequences.</p>
</sec>
<sec id="s4_6">
<label>4.6</label>
<title>Balancing agricultural benefits versus ecological risks: the impact of neonicotinoids on pollinators and the environment</title>
<p>As systemic insecticides, neonicotinoids offer advantages such as high efficacy, targeted action, and ease of application (<xref ref-type="bibr" rid="B92">Simon-Delso et&#xa0;al., 2015</xref>). Their use as seed treatments reduces the need for repeated foliar spraying, lowering labor costs, and farmwork exposure. They effectively combat pests resistant to older insecticides like organophosphates and pyrethroids (<xref ref-type="bibr" rid="B113">Venkatesan et&#xa0;al., 2022</xref>). This long-term crop protection benefits crops like sugar beet, which is often harvested before flowering, reducing direct pollinator exposure (<xref ref-type="bibr" rid="B71">Odemer et&#xa0;al., 2023</xref>).</p>
<p>However, despite their agricultural advantages, neonicotinoids contribute significantly to environmental contamination due to their systemic nature, high water solubility, and persistence in soils and waterways (<xref ref-type="bibr" rid="B98">Stehle et&#xa0;al., 2023</xref>). Their persistence in soil, sometimes exceeding 1,000 days, leads to accumulation and prolonged contamination (<xref ref-type="bibr" rid="B11">Bonmatin et&#xa0;al., 2015</xref>). They have been detected in non-crop plants like wildflowers at field margins, exposing pollinators even in untreated areas (<xref ref-type="bibr" rid="B99">Stewart et&#xa0;al., 2014</xref>).</p>
<p>Some argue that banning neonicotinoids may increase reliance on older, more toxic insecticides, which could pose greater risks to birds, mammals, and fish (<xref ref-type="bibr" rid="B10">Blake, 2018</xref>). Others argue that their routine use as seed coatings contradicts integrated pest management principles, which advocate targeted applications based on pest presence (<xref ref-type="bibr" rid="B37">Goulson, 2013</xref>). The phenomenon of &#x201c;delayed toxicity&#x201d; suggests that lethal effects may only appear after prolonged exposure, complicating environmental risk assessments (<xref ref-type="bibr" rid="B83">S&#xe1;nchez-Bayo and Tennekes, 2020</xref>). While their agricultural benefits are clear, balancing food security with pollinator conservation requires ongoing research into sustainable pest management solutions that minimize ecological harm while ensuring adequate crop protection.</p>
</sec>
<sec id="s4_7">
<label>4.7</label>
<title>Limitations and future directions</title>
<p>Despite adhering to PRISMA guidelines, this review acknowledges several limitations. Inconsistent reporting, especially regarding publication bias assessment, complicates the review evaluation process (<xref ref-type="bibr" rid="B64">Moher et&#xa0;al., 2009</xref>). It is also important to acknowledge that this review is based exclusively on Scopus and Web of Science studies.</p>
<p>The main limitation of neonicotinoid research on bumblebees result from variation in experimental design (lab vs. field), dose ranges, and exposure durations. Together, this leads to results that are not easily comparable or generalizable. Lab studies provide mechanistic insights but lack realism, whereas field studies provide realism but lack control. Future research should prioritize field-realistic exposures and incorporate more naturalistic or semi-field approaches to reflect real-world conditions (<xref ref-type="bibr" rid="B57">Lundin et&#xa0;al., 2015</xref>; <xref ref-type="bibr" rid="B49">J&#xfc;tte et&#xa0;al., 2023</xref>). While lab-based studies permit easy replication and limit confounding factors, doses may not reflect real-world exposure and lack natural variation in diets and population dynamics. This may over- or underestimate the risk depending on experimental setup.</p>
<p>The strong geographical bias toward the UK and the U.S. limits global relevance. This may, in part, be due to restriction to English language only studies. Regions with high bumblebee diversity, such as East Asia and South America, remain underrepresented, constraining the global applicability of existing findings.</p>
<p>Taxonomic bias is another key limitation to understanding the impacts of neonicotinoids on bumblebees (<xref ref-type="bibr" rid="B49">J&#xfc;tte et&#xa0;al., 2023</xref>). The predominance of commercially important species, such as <italic>Bombus terrestris</italic> and <italic>Bombus impatiens</italic>, may overlook responses in other species with different sensitivities or ecological roles, which differ in genetics, stress tolerance, foraging ecology, and disease load. Limited species coverage and inconsistent endpoints make it challenging to draw robust conclusions about population-level risks; therefore, broader species inclusion will be necessary for accurate risk assessments.</p>
<p>The focus of most studies on the three main neonicotinoids may neglect the potential effects of lesser-studied compounds. Future research should assess the safety of alternative neonicotinoids, notably those considered lower risk, to support more sustainable pest management strategies.</p>
<p>Finally, although this review categorized observed effects, it did not quantify exposure thresholds. Establishing dose&#x2013;response relationships is critical for policy development. Future studies should prioritize field-realistic exposures, long-term monitoring, species-specific responses, and broader geographic and climatic coverage. Despite logistical and methodological challenges, these steps are essential for a comprehensive and ecologically relevant understanding of neonicotinoid effects.</p>
</sec>
</sec>
<sec id="s5" sec-type="conclusions">
<label>5</label>
<title>Conclusion</title>
<p>Neonicotinoids are widely applied systemic insecticides, with increasing evidence suggesting toxic effects on non-target organisms including pollinators. This systematic review identified 52 primary studies published within the last 20 years which investigate the effects of neonicotinoids on bumblebees. Key trends were identified, such as geographical and taxonomic biases and commonly reported neonicotinoid compounds and effects. The findings consistently showed sublethal effects, such as impaired foraging, learning, memory, and reproduction, all of which compromise colony health and long-term population viability. Physiological disruptions, including neurotoxicity and immune stress, were also evident. However, the limited focus on pollination efficiency highlights a critical gap in understanding the broader ecological implications of neonicotinoid exposure. Despite regulatory bans and restrictions, debate continues over the effectiveness of neonicotinoid compounds. This review emphasizes the need for expanded research into alternative pest management strategies that balance agricultural productivity and pollinator conservation. With rising global food demands and increasing pest pressures, addressing current research gaps will be vital to developing ecologically sustainable agricultural systems that protect pollinators like bumblebees.</p>
</sec>
</body>
<back>
<sec id="s6" sec-type="data-availability">
<title>Data availability statement</title>
<p>The original contributions presented in the study are included in the article/<xref ref-type="supplementary-material" rid="SM1">
<bold>Supplementary Material</bold>
</xref>. Further inquiries can be directed to the corresponding author.</p>
</sec>
<sec id="s7" sec-type="author-contributions">
<title>Author contributions</title>
<p>DD: Methodology, Conceptualization, Writing &#x2013; original draft, Writing &#x2013; review &amp; editing, Investigation, Visualization, Formal analysis, Data curation. AG: Funding acquisition, Supervision, Project administration, Writing &#x2013; review &amp; editing, Visualization, Conceptualization, Validation.</p>
</sec>
<sec id="s8" sec-type="funding-information">
<title>Funding</title>
<p>The author(s) declare financial support was received for the research and/or publication of this article. This research was performed by DD as part of her final research project for BSc (Hons) in Environmental Science at the University of Nottingham. AG was supported by the BBSRC (grant number BB/Y513866/1).</p>
</sec>
<ack>
<title>Acknowledgments</title>
<p>We wish to thank Dr Reinhard St&#xf6;ger for critical feedback on the manuscript and the two reviewers for constructive feedback during the peer review process.</p>
</ack>
<sec id="s9" sec-type="COI-statement">
<title>Conflict of interest</title>
<p>The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.</p>
</sec>
<sec id="s10" sec-type="ai-statement">
<title>Generative AI statement</title>
<p>The author(s) declare that no Generative AI was used in the creation of this manuscript.</p>
<p>Any alternative text (alt text) provided alongside figures in this article has been generated by Frontiers with the support of artificial intelligence and reasonable efforts have been made to ensure accuracy, including review by the authors wherever possible. If&#xa0;you identify any issues, please contact us.</p>
</sec>
<sec id="s11" sec-type="disclaimer">
<title>Publisher&#x2019;s note</title>
<p>All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.</p>
</sec>
<sec id="s12" sec-type="supplementary-material">
<title>Supplementary material</title>
<p>The Supplementary Material for this article can be found online at: <ext-link ext-link-type="uri" xlink:href="https://www.frontiersin.org/articles/10.3389/frbee.2025.1657493/full#supplementary-material">https://www.frontiersin.org/articles/10.3389/frbee.2025.1657493/full#supplementary-material</ext-link>
</p>
<supplementary-material xlink:href="Table1.docx" id="SM1" mimetype="application/vnd.openxmlformats-officedocument.wordprocessingml.document"/>
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