# Is antibiotic (mis)use an emerging ecological threat to wild insects?

C. Ruth Archer, Jana Dobelmann, Vincent Doublet, Svea Lindeboom, Luisa Linke, Lena Wilfert

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C. Ruth Archer, Jana Dobelmann, Vincent Doublet, Svea Lindeboom, Luisa Linke, et al.. Is antibiotic (mis)use an emerging ecological threat to wild insects?. Journal of Insect Physiology, 2025, 164, pp.104837. 10.1016/j.jinsphys.2025.104837. hal-05154488

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# Is antibiotic (mis)use an emerging ecological threat to wild insects?

C. Ruth Archer<sup>a,* ID</sup>, Jana Dobelmann<sup>a</sup>, Vincent Doublet<sup>a,b</sup>, Svea Lindeboom<sup>a</sup>, Luisa Linke<sup>a</sup>,
Lena Wilfert<sup>a</sup>

## ARTICLE INFO

### Keywords:

- Antibiotics

- Dose Dependence

- Ecotoxicology

- Mate Choice

- Vector-borne Disease

## ABSTRACT

Antibiotic discovery revolutionized human healthcare and boosted agricultural productivity. The efficacy of this revolution is now being challenged however, as the release of antibiotic residues into soils and waterways promotes the evolution and spread of antibiotic resistance genes. There are signs that this antibiotic pollution also has ecological costs, including reduced insect health. Here, we assess this risk. We discuss where, when and how wild insects are exposed to antibiotics, the consequences of this exposure and crucially, if the concentrations that wild insects encounter are sufficiently high to trigger phenotypic responses. Data demonstrate that antibiotic residues reach concentrations in the field that can have phenotypic impacts in insects. These impacts include reduced health and foraging activity in pollinators, and improved survival in insect vectors of disease, demonstrating potential consequences for food-security and disease transmission. More generally, antibiotics can reduce insect stress resistance meaning that antibiotic pollution could elevate insect susceptibility to other stressors implicated in insect declines. We highlight gaps in our understanding of how antibiotic pollution affects wild insects and the ecosystem services they provide. These gaps urgently need to be filled, because global antibiotic use is rising. Crucially, we must determine how antibiotic residues in the field degrade or accumulate along food chains, and how field-realistic concentrations affect insects directly and via interactions with other environmental stressors. This will offer concrete insights into the consequences of rising antibiotic use for insects, and their broader societal and ecological impacts.

## 1. Introduction

Many wild insect populations are in decline (Wagner, 2020). These declines are driven by multiple stressors (e.g. land-use and climate change, pollution, insecticide use and nitrification) (Wagner, 2020). These stressors can interact such that even if each has a modest fitness effect, their combined impacts can be stark (discussed in Tekin et al., 2020). Understanding how environmental stressors affect insects, both singly and in combination, is therefore central to predicting and mitigating insect declines in the Anthropocene. To this end, research has explored the effects of environmental toxins (e.g. pesticides) on non-target insects (Aliouane et al., 2009; Boff and Ayasse, 2024; Human et al., 2014; Schneider et al., 2012) and tested how these interact with other factors such as diet quality (Archer et al., 2014; Leza et al., 2018) or pathogens (Doublet et al., 2015) to affect phenotype. One emerging class of pollutant whose toxic effects on wild insects are only beginning to become clear, is antibiotics.

By allowing the treatment of bacterial pathogens and enabling medical interventions such as organ transplants and open-heart surgeries, antibiotics have helped fuel human life-expectancy increases of > 20 years in the last century (Hutchings et al., 2019). Antibiotics are also used in farms and in aquaculture to manage and prevent outbreaks of plant and animal diseases and to increase livestock feed conversion efficacy (Kirchhelle, 2018). Accordingly, antibiotic discovery heralded rapid improvements in human health and agricultural productivity. Wide-scale antibiotic use means however, that antibiotics are increasingly ending up where they do not belong and entering natural ecosystems. Low-level leaching of antibiotics into ecosystems can select for the evolution of antibiotic resistance genes and enhance their spread. This is a tremendous concern because antimicrobial resistance is a growing threat to human health (Murray et al., 2022) and food security (Haulisah et al., 2021). In addition, antibiotic pollution might affect non-target microbes (Grenni et al., 2018) and multi-cellular organisms. This pollution could therefore, in theory at least, have costly impacts on

<footnote><sup>a</sup> Institute of Evolutionary Ecology and Conservation Genomics, University of Ulm, Albert-Einstein Allee 11, D-89069 Ulm, Germany</footnote>

<footnote><sup>b</sup> DGIMI, Univ Montpellier, INRAE, Montpellier, France</footnote>

<footnote>* Corresponding author.</footnote>

<footnote>E-mail address: rutharcher0@gmail.com (C.R. Archer).</footnote>
wild insects.

There is clear potential for antibiotic pollution to reduce wild insect fitness or elevate their vulnerability to other stressors. This is because antibiotics induce gut dysbiosis (perturbed gut microbial communities), which disrupt the functional roles performed by gut bacteria and in doing so, could affect insect chemical communication (Engl and Kaltenpoth, 2018), reproductive mode (Stouthamer et al., 1990), behaviour (Lewis and Lizé, 2015) and life-history (Macke et al., 2017). Antibiotics can also have direct toxic impacts on insect hosts (Ridley et al., 2013); not least because the attributes that make antibiotics lethal to bacterial pathogens also make antibiotics dangerous to host mitochondria (Wang et al., 2015). Whether these costly impacts on insects are realized in the field however, is unclear.

The aim of this review is to ask if antibiotics are an emerging ecologically relevant stressor that could contribute to insect loss. Answering this question requires understanding 1) where, when and how terrestrial and aquatic insects are exposed to veterinary and medical antibiotics, 2) the potential consequences of antibiotic exposure and crucially, 3) if the concentrations that wild insects encounter are high enough to trigger phenotypic responses. We do not tackle the role of insects in the evolution and spread of antimicrobial resistance because this has been discussed elsewhere (e.g. Rawat et al., 2023).

## 2. Where, when and why do wild insects encounter antibiotics?

### 2.1. Potential spillover from deliberate antibiotic application

Insects perform a suite of ecosystem services including pollination, biocontrol and recycling of organic matter and can serve as a source of human food and animal feed. Harnessing these services often involves rearing large numbers of animals at high densities, creating an ideal environment for bacterial disease spread (Greenberg, 1970). Thus, antibiotics are sometimes used to manage bacterial infections in mass-rearing contexts. In some sectors antibiotics are used sparingly because of negative impacts on insect health and the risk of antibiotic resistance emergence and spread (Grau et al., 2017). For example, antibiotics are used to manage bacteriosis outbreaks in insects reared en masse for food and feed (EFSA Scientific Committee, 2015), but in general antibiotic use is avoided in this sector (Mitchaothai et al., 2024). Some insects reared for feed are exposed to antibiotic residues none-the-less. Black soldier fly (Hermetia illucens) larvae for example, are often reared on manure, which they convert into protein and fat (Barragan-Fonseca et al., 2017). Manure can contain antibiotic residues (Oliver et al., 2020) and research is ongoing to understand how this antibiotic exposure affects black soldier flies and how readily these insects degrade antibiotic residues (Cai et al., 2018; Liu et al., 2020).

While certain industries avoid antibiotic use, others appear to embrace it. Sericulture, the mass rearing of silkworms (Bombyx mori) for silk, is an important income source in some rural communities (Ekka and Bais, 2023). Sericulture is billed as being eco-friendly because silk production can be incorporated into circular economies e.g. waste from silk moth rearing can be used as compost (Altman, 2022). However, some papers advocate the use of antibiotics for improved stock health and silk production (Mahdi et al., 2017; Rafiq et al., 2020) and suggest that antibiotic use is widespread in the industry (Rafiq et al., 2021). Apiculture is another sector where antibiotic use is common. In the USA honey bee hives are treated with antibiotics to protect against costly and virulent diseases such as foulbrood and nosemosis (Forsgren, 2010; Marín-García et al., 2022). Long-term antibiotic exposure in American bees has caused honey bee microbiomes to accumulate antimicrobial resistance genes (Tian et al., 2012) and probiotic use is being trialed to mitigate some of the negative impacts of this antibiotic exposure on bee health (Motta et al., 2022).

While sericulture and apiculture use antibiotics to improve the health of their insect stocks, the pest management sector use antibiotics to different effect. Some insects that damage crops or vector diseases,

host bacterial symbionts that are beneficial (or even essential) to host fitness. For example, sap feeding silverleaf whiteflies (Bemisia tabaci) rely on *Portiera* bacteria for carotenoid biosynthesis (Sloan and Moran, 2012) and detoxifying pathways in the brown planthopper (*Nilaparvata lugens*) are connected to bacterial endosymbionts (Tang et al., 2021). Removing these endosymbionts via antibiotics can reduce the fitness of insect pests and make them more vulnerable to other pest management strategies, including pesticides (Rupawate et al., 2023). While there is recognition that antibiotic use has risks in terms of non-target organism effects and antibiotic resistance evolution (Qadri et al., 2020; Zhang et al., 2022), some researchers have advocated highly targeted antibiotic application as part of wider integrated pest management program (Zhong et al., 2007).

This deliberate antibiotic use illustrates a potential problem: treating certain insects with antibiotics may bring antibiotics into contact with non-target species. In the case of sericulture, antibiotic use (Rafiq et al., 2021) coupled with efforts to build silk production into circular economies might result in accidental exposure of non-target insects e.g. where silkworm excrement and breeding waste are used as compost (Altman, 2022). Similarly, using antibiotics to kill insect pests may provide a route through which antibiotic treated insects enter the food-chain and natural ecosystems, providing scope for bioaccumulation / biomagnification of antibiotics along food chains. The line between deliberate antibiotic application in managed insects and inadvertent environmental release is therefore blurred. The scale of this problem seems likely to be small however, compared to the effects of accidental leaching of antibiotics used to promote human or animal health.

### 2.2. Accidental antibiotic accumulation in the wild

Antibiotic use in hospitals is high, with ~ 1 in 3 inpatients in European hospitals in 2016-17 being treated with antimicrobials (Plachouras et al., 2018). These medical antibiotics could reach wild insects via different routes. Most directly, blood-feeding insects may consume antibiotics when feeding on a treated individual (Gendrin et al., 2015). In regions where diseases vectored by insects (e.g. malaria, sleeping sickness, Zika) are common, this could be an important pathway of insect exposure to antibiotics with far-reaching impacts on disease dynamics (discussed in Section 4) (Gendrin et al., 2015). Insect pests of hospitals and mortuaries more generally are important vectors of antibiotic resistance, in part due to a propensity to feed on human tissue (Gwenzi, 2020).

Other insects may be exposed to medical antibiotics via less direct routes. Antibiotics that are administered to patients are not fully metabolized (Ngumba et al., 2020; Zhou et al., 2021) and as a result, antibiotic residues are detectable in waste waters even after passage through waste management facilities (Rodriguez-Mozaz et al., 2015). Antimicrobials therefore contaminate rivers such as the Thames (UK) (White et al., 2019) and the Ganges (India) (Lapworth et al., 2018) and reach levels sufficiently high to drive antimicrobial resistance selection (Hayes et al., 2022). This antibiotic pollution provides a route of exposure for insects living in waterways or soils contaminated with wastewater or effluent.

Veterinary antibiotics can reach insects via many of the same routes. Antibiotics can be consumed by insects feeding on (or living alongside) treated animals – perhaps helping explain the high antibiotic resistance observed in flies, lice, fleas and beetles living in or around farms (Rawat et al., 2023). Antibiotics are also excreted by livestock, meaning that agricultural fields can be contaminated by veterinary antibiotics via the application of manure as a fertilizer, or waste water for irrigation (Pan and Chu, 2017). Once more, this could bring soil dwelling insects into antibiotic contact, and run-off and waste water could expose insects with aquatic life-stages (e.g. damselflies, mosquitoes) to antibiotics. Antibiotics in soil may then be taken up by crops (Zhao et al., 2019), potentially exposing herbivorous insects to antibiotic residues. In aquaculture, the route between treatment and contamination of waterways is even
more direct: around 80 % of the antibiotics deployed to fresh-water fish populations disperse into waterways (Cabello et al., 2013), polluting rivers and sediments in turn (Sargenti et al., 2020).

Agricultural antibiotic use is not restricted to livestock; antibiotics are also sprayed onto crops (Taylor and Reeder, 2020). Fireblight for example, a virulent disease of apples and pears, is treated with antibiotics in many parts of the world (Haynes et al., 2020). This treatment is effective when sprayed onto plant blossoms and so occurs in spring (Stockwell and Duffy, 2012). Accordingly, fireblight treatment coincides with pollination, meaning that pollinators may consume antibiotics alongside pollen and nectar.

In summary, blood-feeding insects have a clear route of exposure to veterinary and medical antibiotics and other insects may be exposed to antibiotic residues that trickle into our rivers, streams, and soils. Bio-accumulation of antibiotics from soils into plant tissues, and direct application onto flowering crops, may expose insect herbivores to antibiotics as well (Fig. 1). This is a clear and urgent problem from the perspective of antibiotic resistance evolution (Rawat et al., 2023). But, does antibiotic pollution also pose a threat to insect species and the ecosystem services they provide? For this to be the case, antibiotic exposure must have costly phenotypic impacts on insect hosts. In support of this, there is abundant evidence from lab experiments that antibiotics reduce fitness, reshape life-histories and disrupt behaviour in a broad range of insects. In the following sections, we review some of these impacts.

### 3. How might accidental antibiotic exposure affect insects?

There are many examples of how antibiotics can affect insect phenotype. Rather than trying to document the full spectrum of these effects, we demonstrate that antibiotics can affect multiple aspects of individual insect fitness and potentially exacerbate the impacts of other stressors that are driving insect losses. To provide this summary we focus

on three categories of fitness impacts: life-history, behaviour and stress resistance. The references cited in this section are summarized in more detail (i.e. which antibiotic classes are tested and at what concentrations) in Table S1. While we do not discuss the mechanisms responsible for any antibiotic impacts on phenotype in detail, broadly there are two candidate pathways. First, antibiotic exposure can disrupt insect gut microbiota (i.e. induce dysbiosis) and, because gut microbial communities play crucial functional roles (e.g. nutrient processing, protection against pathogens, synthesis of pheromones etc) (Engl and Kaltenpoth, 2018; Moran et al., 2019), this disruption could reduce insect health. Second, antibiotics can have direct toxic effects on hosts by interfering with mitochondrial function or by inhibiting enzymatic detoxification pathways (Renault et al., 2018) (Fig. 2). We often lack the data needed to tease apart these possibilities, which is why a discussion of mechanisms is beyond the scope of this review.

#### 3.1. Antibiotic effects on life-history

Whether an individual reaches maturity, survives to have one (or more) reproductive opportunities and can use these opportunities (i.e. transfer viable sperm etc.) are critical determinants of individual fitness. In insects, antibiotics can affect each of these vital life-history traits i.e. growth, survival and reproduction.

Exposure to antibiotics can reduce the likelihood that individuals survive to adulthood, as seen in common green bottle fly (Lucilia sericata, previously Phaenicia sericata) larvae fed high antibiotic concentrations (Sherman et al., 1995) and developing Drosophila melanogaster fed a range of penicillin doses (Üstündağ et al., 2020). Juveniles fed antibiotics can also grow more slowly, an effect seen in whiteflies (Bemisia argentifolii) (Costa et al., 1997), houseflies (Musca domestica) (Li et al., 2021), the black bean aphid (Aphis fabae) (Jayaraj et al., 1967), German cockroaches (Blattella germanica) (Liles, 1958) and wax moths (Galleria mellonella) (Büyükgüzel and Kalender, 2008). In contrast, antibiotic

<figure><img src="image_3.png" /><figcaption>**Fig. 1.** A schematic illustrating possible pathways linking veterinary and medical antibiotics with non-target, wild insects. Insects circled in orange demonstrate direct exposure routes (i.e. feeding on antibiotic treated people, livestock, crops or waste), while insects circled in purple represent possible indirect routes of exposure (i.e. via leaching into waterways and soil, and uptake by crops). Arrows show potential flow of antibiotics into waterways and soils. A) Direct blood feeding on treated patients, B) release of incompletely metabolized antibiotics into waterways, C) possible leaching from degrading cadavers, D) and leaching from water to soil, E) foraging on antibiotic treated flowering crops, F) blood feeding on treated livestock, G) consumption of antibiotic containing manure, H) leaching from manure into soils, I) uptake (and possible accumulation / magnification) of antibiotics from manure treated soils into crops. Insects shown are representatives of the communities and life stages that may be found in contaminated habitats, or that may directly encounter antibiotics via the routes highlighted. Figure was made in BioRender by CRA.</figcaption></figure>
<figure><img src="image_4.png" /><figcaption>**Fig. 2.** Example routes through which antibiotics can disrupt host health. Broadly, antibiotics can affect host phenotype directly (blue – via toxicity) or indirectly (purple – via gut dysbiosis). Effects of gut dysbiosis are likely to be characterized by complex interactions, for example, disruption of the gut microbiota affects nutrient metabolism, which in turn can affect thermal tolerance, production of insect pheromones etc.</figcaption></figure>

treated wood tiger moth (Arctia plantaginis) larvae grow faster than controls, but develop into smaller adults (Galarza et al., 2021). Juvenile insects fed antibiotics often show reductions in their size during development and / or when they emerge as adults (Büyükgüzel and Kalender, 2008; Jayaraj et al., 1967; Li et al., 2021; Lin et al., 2015) and may display morphological defects (Shabbir et al., 2024) or greater asymmetry – an indicator of developmental stress (Büyükgüzel et al., 2020). Antibiotic impacts on juveniles can roll over from one life stage to the next; as seen in female *Culex pipiens* mosquitoes who survive better as adults when exposed to antibiotics as juveniles (Garrigós et al., 2024). Impacts of antibiotic treatment on juveniles can also roll over from one generation to another, as in German cockroaches where individuals whose parents were treated with antibiotics grew into smaller adults (Liles 1958).

While there are many examples of antibiotic treatment affecting ju-
venile insects (Table S1), antibiotic effects can vary across insect species
and even between strains of the same species (Stevens and Wade, 1988).
Responses often depend on the type of antibiotic used; in bottle flies for
example, survival or maturation were reduced in larvae that consumed
gentamicin and cefazolin but not those fed ampicillin, ceftizoxime,
clindamycin, mezlocillin, or vancomycin (Sherman et al., 1995). Simi-
larly, the impacts of antibiotics are often dose-dependent; these negative
impacts of gentamicin and cefazolin in bottle fly larval survival were
only seen at high concentrations (Sherman et al., 1995). In other words,
the relationship between antibiotics and larval development and sur-
vival is complicated.

Antibiotics have similarly striking effects on adult life-history traits.
Adult antibiotic exposure can reduce survival in the walnut husk fly
*Rhagoletis completa* (Tsiropoulos, 1981), eastern subterranean termites
*Reticulitermes flavipes* (Sapkota et al., 2021) and bumble bees *Bombus
terrestris* (Marceau et al., 2021). Female reproductive success can also be
reduced by antibiotic treatment. Female egg laying was reduced by 97 %
in black bean aphids treated with antibiotics as larvae (Jayaraj et al.,
1967) and cockroach females treated with antibiotics across their
development produced smaller and misshapen eggs (Liles, 1958). An-
tibiotics reduce hatching success and fertility in *Aedes albopictus*
mosquitos (Ha et al., 2021). In *D. melanogaster*, reductions in offspring
eclosion were observed following antibiotic treatment of adults, due to
antibiotic effects on mothers rather than fathers (Singh et al., 2024).
Once more, effects of antibiotics on female fecundity can carry over from
one generation to the next; the fecundity of granddaughters of antibiotic
treated *Tribolium confusum* beetles was elevated relative to controls (Li
et al., 2016). Additionally, effects of antibiotics on female fecundity can
depend on interactions with other factors such as diet quality (Goane
et al., 2022).

Male reproductive success is hard to measure because it relies on a
complex mixture of traits (Archer et al., 2022); these include how well
males court females and outcompete rivals to secure mates, produce
large numbers of functional sperm and seminal fluid, and invest in traits
that secure success in sperm competition / cryptic female choice (e.g.
nuptial gifts, copulatory courtship). There are signs that some of these
attributes can be influenced by antibiotic treatment. Antibiotics alter
male cuticular hydrocarbon (CHC) profiles – a lipid layer covering the
bodies of insects that help prevent desiccation and can act as contact
pheromones involved in mate choice. In tsetse flies (Glossina morsitans
morsitans) for example, maternal antibiotic treatment affected male and
female offspring CHC profiles and these offspring were less frequently
selected as mates (Engl et al., 2018). Antibiotic disruption of sex pher-
omones has also been observed in the true fruit flies *Anastrepha ludens*
(Aluja et al., 2020) and *Bactrocera dorsalis* (Ren et al., 2021) and flour
beetles *Tribolium castaneum* (Wang et al., 2024a), while negative effects
of antibiotics on male attractiveness have been observed in
*D. pseudoobscura* (Heys et al., 2020). As to how antibiotics affect male
fertility *per se* is less clear. For sperm storage for artificial insemination,
Poole and Taber (1970) found that the addition of antibiotics to honey
bee (*Apis mellifera*) semen improved sperm survival at low temperatures
but later work found no impact of antibiotics on honey bee sperm
viability (Andere et al., 2011).

3.2. Antibiotic effects on behaviour

In addition to affecting insect life-histories, antibiotic intake can
shape insect behaviour. These effects have been particularly well studied
in bees. Antibiotics reduce learning and foraging success in bumble bees
(Avila et al., 2022) and honey bees foraging in pear orchards treated
with antibiotics visit fewer flowers (Avila et al., 2024). Given that
foraging success is vital to colony fitness, these impacts on bee behav-
iour may have costly impacts that ripple out across the colony. In honey
bees the usual progression in behaviours displayed by workers (age
polyethism) is also disrupted by antibiotic treatment (Ortiz-Alvarado
et al., 2020). This division of labour is vital to colony cohesion and
success, meaning that once more, antibiotic induced behavioural
disruption could have costly and far-reaching impacts on a social insect
colony. At scale, this behavioural disruption could, in theory, reduce
agricultural production by reducing pollination efficiency – whether this
is likely, is a question that we return to.

Antibiotic treatment can also shape insect communication. One
important route for this is that antibiotics can disrupt insect pheromones
including CHCs, whose production often relies on insect microbial
communities (Engl and Kaltenpoth, 2018). In honey bees, antibiotic
treatment affects worker CHCs involved in nest-mate recognition (Vernier et al., 2020). Antibiotics also affect CHC profiles in the bumble bee *B. terrestris*, and in particular, affect the pheromones that queens use to signal their reproductive dominance and suppress reproduction in workers (Archer et al. in prep). In vinegar flies (*D. melanogaster*) anti-biotic treatment can affect aggregation behaviours; larvae treated with Rifampicin lose their ability to discriminate against foods that are already occupied by larvae (Thibert et al., 2016).

As suggested above, antibiotics can affect female mate choice. Antibiotic treatment led to reduced mate discrimination in subsequent generations of *D. melanogaster*, an effect attributed to *Wolbachia* removal (Koukou et al., 2006). Antibiotic treatment similarly altered female mating preferences in *D. subobscura* (Walsh et al., 2017). By relaxing female choosiness and disrupting female mating preferences, antibiotics could lead to females accepting more deviation from their preferred mate phenotypes and so diminish any fitness benefits of mate choice. Crucially, these effects show that antibiotics could disrupt a key mechanism of sexual selection (Andersson, 1994; Archer and Hosken, 2021).

### 3.3. Antibiotic effects on stress resistance

Evidently, antibiotic treatment can affect behaviours and life-history traits that influence individual fitness in benign lab conditions. In the field, conditions are seldom benign and instead contain a suite of stressors. Antibiotics can disrupt how well insects resist stressors, including those implicated in driving insect declines such as pollution, nutritional deficiencies through habitat loss, extreme weather events and pathogens.

In a range of insects, antibiotic treatment reduces detoxification capacities. The ability of coffee berry borers (*Hypothenemus hampei*) to degrade caffeine (a toxic alkaloid) is removed by antibiotic treatment (Ceja-Navarro et al., 2015). Antibiotics elevate vulnerability to pesticides in species including the brown planthopper *N. lugens* (Tang et al., 2021; Zhang et al., 2021), the Asian citrus psyllid *Diaphorina citri* (Pan et al., 2024), and the honey bee *A. mellifera* (Wu et al., 2020). These impacts appear to be largely due to the removal / reduction of bacterial symbionts with key roles in detoxification (reviewed in Itoh et al., 2018; Peterson, 2024). These effects demonstrate the complexity of antibiotic impacts on wild insects: in the pest-management literature, these negative impacts of antibiotics on insect detoxification capacities are valuable because they elevate pesticide efficacy. These same effects on beneficial non-target organisms, including pollinating bees, can have costly ecological, social and economic impacts. Moreover, pesticide pollution can select for pesticide resistance in bacteria, which can result in the acquisition of antimicrobial resistance as an additional trait (Ramakrishnan et al., 2019). Evidently, complex interactions between toxins and antibiotics, mediated by host microbiota, can have deleterious impacts on beneficial insects and on the evolution of antimicrobial resistance.

Antibiotics may also mediate the impacts of thermal stress. In the red palm weevil (*Rhynchophorus ferrugineus*), antibiotic treatment reduces survival at high and low temperatures (Elkraly et al., 2024). In the true fruit fly *B. dorsalis*, antibiotic treatment reduced the median survival of flies at low temperatures from 19 days to 6 days (Raza et al., 2020), with similar results in *C. pipiens pallens* mosquitoes (Lv et al., 2023). In these cases, effects appear to be due to reductions in key metabolites linked to gut microbiota. Given that climatic change is an increasingly influential driver of insect declines (Blüthgen et al., 2023), antibiotic impacts on thermal stress resistance highlight another route through which antibiotics can increase insect vulnerability to other important environmental stressors.

By reducing immunocompetence (Galarza et al., 2021), antibiotics can contribute to the spread of infectious diseases. In codling moths *Cydia pomonella*, antibiotics reduce the expression of antimicrobial peptides and in turn, elevate individual vulnerability to pathogens (Li et al., 2024). In honey bees, oxytetracycline treatment reduced key

symbionts (e.g. *Frischella perrera*, *Lactobacillus Firm-5*) known to regulate immune function and nutrient metabolism, and thus reduced the antimicrobial capacity of worker haemolymph (Daisley et al., 2020). Antibiotic treated honey bees are also more likely to acquire infections by the microsporidian *Nosema* (Li et al., 2017), while in bumble bees, antibiotic treated animals are more likely to become infected with the gut parasite *Crithidia* (Marceau et al., 2021). Emerging infectious diseases pose a tremendous threat to biodiversity (Strauss et al., 2012), and so such negative impacts on immunity provide one more way that exposure to antibiotics could threaten wild insect populations.

Finally, antibiotics can exacerbate malnutrition by removing the gut microbiota that help extract nutrients from hard to digest foods and provision hosts with metabolites. In bees for example, gut bacteria help break down hard to digest foods such as pollen (Engel et al., 2012; Kešnerová et al., 2017). Presumably due to this, antibiotic exposure impairs protein digestive efficiency in African honey bees (*A. mellifera* *scutellata*), especially in individuals fed low-protein diets (Du Rand et al., 2020). Given that the amount and ratio of nutrients that individuals consume affects multiple fitness traits including survival, immunity, female fecundity, male attractiveness and fertility (Bunning et al., 2016; Malod et al., 2017; Ponton et al., 2013), effects of antibiotics on nutrition are likely to affect multiple fitness traits.

### 4. Impacts in the field – A question of concentration

Antibiotics have the potential to reduce multiple aspects of insect fitness directly (i.e. reduced fertility or survival) and indirectly (i.e. elevated vulnerability to other stressors / disrupted sexual selection). By altering insect growth, reproduction and survival in the field, antibiotics could alter insect population dynamics. However, many antibiotic residues that reach natural ecosystems will be diluted, degraded, bound in soil or taken up by plants and thus, exhibit lower bioavailability (Cycoń et al., 2019). This raises the question, are antibiotic concentrations in the field of any real concern to insect health?

This is not an easy question to answer because antibiotics frequently show dose-dependent effects. Antibiotic impacts may rise linearly across a concentration gradient (Fig. 3A), either immediately or only above a threshold dose (Fig. 3B). More complex hormetic responses, where low doses have a positive effect on fitness and high doses reduce fitness, are also possible (Fig. 3C). For example, in 4th instar wax moths (*G. mellonella*) low doses of penicillin reduced markers of oxidative damage to lipids, while high doses elevated them (Büyükgüzel and Kalender, 2014). Such complex species, stage and dose specificity in outcomes makes it impossible to draw general conclusions about how antibiotics affect insect populations. However, in some species, ecologically relevant antibiotic concentrations are having striking phenotypic impacts, which may in turn affect human health and food security.

Crucially, mosquitoes fed blood of antibiotic treated humans experienced gut dysbiosis (Gendrin et al., 2015). These mosquitoes lived longer, were more fecund and more likely to acquire malarial infection. Antibiotic exposure therefore had positive effects on mosquitoes, that could facilitate the transmission of malaria (Gendrin et al., 2015). The microbiome of mosquitoes also mediates the transmission of arboviruses (arthropod-borne viruses), with specific bacteria either improving or reducing host vector competence (Hegde et al., 2015). *Wolbachia* for example can enhance West Nile virus replication inside mosquito cells, but inhibits virus assembly or secretion (Hussain et al., 2013). Ecologically realistic doses of antibiotics found in waste-water reduce *Wolbachia* infections in *Ae. aegypti* (Endersby-Harshman et al., 2019), meaning that antibiotic uptake by mosquitoes as they develop in water bodies, or following blood intake, could affect the spread of arboviruses such as dengue virus, chikungunya virus, and West Nile virus. This suggests that increased vector-control may be needed in areas with high antibiotic use in humans or in livestock farming and demonstrates how antibiotic effects on insects could have costly impacts on human health. Future work determining whether similar impacts are seen in other insect vectors of
<figure><img src="image_5.png" /><figcaption>**Fig. 3.** Potential dose dependent impacts of antibiotics on their hosts. Responses in individuals fed antibiotics are shown as solid lines and are presented relative to the control (i.e. the absence of antibiotics), whose responses are shown as a dashed grey line. Panel A and B show a linear relationship between antibiotics and phenotype without (A) or with (B) a threshold, and Panel C shows a hormetic dose-response relationship. In this example, low-doses reduce and high-doses enhance costly phenotypic responses (e.g. death, disease, oxidative damage etc) (Calabrese, 2003). Figure was made in BioRender by CRA.</figcaption></figure>

disease is urgently needed.

There is also evidence that ecologically relevant antibiotics affect pollination. In lab experiments, reduced learning and health have been observed in bumble bees treated with antibiotic concentrations that are sprayed on crops (Avila et al., 2022; Marceau et al., 2021). Honey bees that visited pear orchards sprayed with antibiotics visited fewer flowers and foraged for longer, showing reduced foraging efficiency (Avila et al., 2024). Here, no effect of this reduced foraging efficiency was found on seed set. However, the focal pear cultivar in this study was self-compatible, and thus not highly pollinator-dependent, and treated only once with antibiotic spray (Avila et al., 2024). In the field, crops are often sprayed multiple times (Walgenbach et al., 2012) and so the potential for antibiotic induced reductions in visitation by insect pollinators to impact pollination services remains.

Antibiotic concentrations in manure can reach high levels (Oliver et al., 2020) meaning that any insects feeding on manure may consume high antibiotic doses. In support of this, gut microbiota were disturbed in dung beetles fed manure from cattle treated with antibiotics, although this did not affect dung beetle size, survival or reproduction (Hammer et al., 2016). Similarly, in black soldier flies used in manure management high doses (10 mg/kg) of sulfonamides (Gao et al., 2019) and oxytetracycline (5 mg/kg) impaired larval development (van Dongen et al., 2024). These levels can be exceeded in livestock manure (Alavi et al., 2015; Martínez-Carballo et al., 2007). Once more, these results illustrate the complexity of this topic: whether insects that consume manure are likely to accumulate antibiotics at levels high enough to affect insect phenotype depends on the insect, the antibiotic class and dose that is applied – factors that vary geographically and in relation to local agricultural practices.

In the case of insects that consume blood or manure from antibiotic treated livestock, or forage on antibiotic sprayed crops, the pathway between antibiotic treatment and insect consumption is short and direct. These groups are strong candidates for being affected by antibiotics in the field. What about insects where the routes of antibiotic exposure are longer and indirect, such as aquatic insects in antibiotic polluted waterways or soil dwelling insects in manure treated fields? Here, the impacts of exposure on insects are even less clear. None-the-less, some antibiotic classes can reach high concentrations in soil due to repeated fertilizations with liquid manure (Hamscher et al., 2002) and this can affect the abundance of soil dwelling invertebrates such as earthworms (Zhao et al., 2022). Levels of antibiotics detected in waterways can also reduce insect fitness, as highlighted above, in *Ae. aegypti* (L.) mosquitoes a high (but potentially field realistic) concentration of antibiotics slowed larval growth (Endersby-Harshman et al., 2019). Antibiotic pollutants accumulated in wild insects collected from a Chinese river delta (Hu et al., 2023) and models predict that in aquatic systems, sulfonamide

antibiotics can undergo biomagnification, such that their concentrations rise with movement along the food-chain towards higher-trophic levels (Zheng et al., 2025).

## 5. Conclusion

In terms of answering the question this review initially posed, “*are antibiotics an emerging ecological threat to insect populations?*” the answer seems to be – it depends. The impacts of antibiotic pollution depend on local agricultural practices and legislation, as well as on climatic factors that affect how rapidly antibiotics degrade. There will be variation in antibiotic impacts between insects, depending on their susceptibility to specific antibiotic classes and their reliance on beneficial gut bacteria. There will be variation between populations due to interactions between antibiotics and other drivers of insect decline, such as pesticide use. This is a complex puzzle, but amidst this complexity, it is clear that in insects with a clear and direct route to antibiotic exposure, field-realistic antibiotic concentrations can have phenotypic impacts. These effects on insects can be detrimental, as seen in bees foraging in treated orchards, or positive (from the insects’ perspective), as seen in mosquitoes vectoring malaria. Therefore, there is potential for antibiotics to be another ingredient in the complex cocktail of stressors that wild insects are exposed to.

However great the risk posed by antibiotics to insect health is today, this risk is rising. By 2030, global antibiotic consumption is expected to increase by 52.3% from 2023 levels (Klein et al., 2024). Managing the ecological consequences of rising antibiotic use, as well as the growing risk of antimicrobial resistance evolution, requires a better understanding of antibiotic impacts on natural populations. We must determine how much and how quickly antibiotic residues degrade in the field or accumulate as they ascend trophic levels in food chains. We need to understand how field-realistic doses affect wild insects directly and via interactions with the myriad of other stressors wild insects encounter. In particular, research exploring antibiotic-insecticide synergisms would offer tremendous insight into the potential for even low antibiotic concentrations to have costly ecological impacts.

In concert, it is important that we reduce antibiotic use in agriculture and animal husbandry and adopt alternative strategies to reduce disease spread, increase animal health, and treat infections. This can be achieved by flock and herd management strategies that facilitate disease control e.g. hygienic environments and biosecurity measures such as air filtration systems (Kahn et al., 2019). Another way to reduce disease spread is vaccination against common diseases; vaccinations against the bacterial pathogen *Staphylococcus aureus* can reduce mastitis incidence / severity in dairy cows for example (Ismail, 2017). Other alternatives to antibiotics include insect derived antimicrobial peptides (AMPs) and the
addition of insect protein to animal feed. Resistance evolution against AMPs is slower than resistance evolution against antibiotics (Lazzaro et al., 2020) and animals fed with an insect diet showed strengthened immunity and increased antibacterial activity (Patyra and Kwiatek, 2023). Phytogenic feed alternatives (feed containing pharmacological plant components) have been shown to increase growth, performance and health of animals and are less likely to induce resistance evolution (Wang et al., 2024b). Where alternatives to antibiotics are not available, research examining how to minimize the pollution caused by antibiotic use is another important research frontier. Here, as highlighted above, insects may help provide a solution, as species including black soldier flies can degrade some antibiotic residues in manure without accumulating them in their tissues (Gao et al., 2019). Finally, we urge sectors that consider use of antibiotics to manage insect pests or in mass-rearing of beneficial insects, to balance the benefits of doing this against the risk of antibiotic resistance evolution, potential increases in disease vectoring capacity and direct harmful impacts on wild insect populations, including human-beneficial insects such as pollinators.

Finally, in legislation that regulates antibiotic use, increased awareness of potential impacts on wild species, including insects, would be welcome. While the use of streptomycin as a pesticide has been banned in major agricultural producers such as the EU and Brazil (Donley, 2019), the U.S. Environmental Protection Agency (EPA) approved streptomycin use on citrus crops, which has been linked to streptomycin-resistant bacteria in the honey bee gut microbiome (Ludvigsen et al., 2018). Although this approval was recently withdrawn after a court ruled that the EPA had not sufficiently assessed environmental risks, including risks to pollinators (US Court of Appeals, 2023). In the EU, proposals have been made to establish environmental quality standards for antibiotics in surface waters, and some European countries have implemented national guidelines (Agerstrand et al., 2023), yet, there are no legally binding standards. The recent addition of oxytetracycline and tetracycline to the EU surface water watch list as contaminants of emerging concern is expected to improve environmental monitoring (Gomez Cortez et al., 2025) and may support the development of protective standards, particularly for aquatic insects. This is a welcome step in a world where antibiotic use is on the rise and the negative consequences of antibiotic pollution are increasingly clear.

## CRediT authorship contribution statement

C. Ruth Archer: Writing – original draft, Visualization, Conceptualization. Jana Dobelmann: Writing – review & editing, Conceptualization. Vincent Doublet: Writing – review & editing, Conceptualization. Svea Lindeboom: Writing – review & editing, Conceptualization. Luisa Linke: Writing – review & editing, Conceptualization. Lena Wilfert: Writing – review & editing, Conceptualization.

## Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

## Acknowledgements

CRA is funded by the German Research Council (DFG AR 1369/2-1). JD is funded by an incubator grant from the University of Ulm. LL is funded by the German Research Council (DFG DO 2570/2-1 to VD). This study is part of the BeePath project that has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (Grant Agreement No. 864642).

# Appendix A. Supplementary data

Supplementary data to this article can be found online at https://doi.org/10.1016/j.jinsphys.2025.104837.

## Data availability

No data was used for the research described in the article.

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