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Review · Volume 40, Issue 2, 8 December 2025 · Open Access

Assessing the Impact of Pesticides on Earthworm Health: A Comprehensive Review

Affiliations & Notes ⌄
  1. 1Department of Pesticide Chemistry and Technology, Faculty of Agriculture, Alexandria University, Aflaton St., 21545, El-Shatby, Alexandria, Egypt Corresponding author: Mohamed R. Fouad mohammed.riad@alexu.edu.eg, mohammed.reid@yahoo.com
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Punjab University Journal of Zoology
Published:
8 December 2025
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From the article

  • This review underscores the complex and multifaceted threats that pesticides pose to earthworm populations and, consequently, to soil health and ecosystem stability.
  • Pesticides can also induce significant histopathological alterations, including degeneration of the epidermis and gut epithelium, alongside genotoxic effects that compromise cellular integrity and genetic stability, potentially impairing earthworm populations and their ecological functions.
  • By synthesizing these patterns and translating them into targeted actions and research priorities, we can better safeguard earthworm populations, preserve vital ecosystem services, and promote sustainable agricultural practices for future generations.

Selected from the article's own text.

Abstract

This review consolidates recent global research on pesticide production, usage patterns, and their ecological impacts on earthworms, emphasizing their role as bioindicators of soil health. The main objective is to identify key pesticide groups and biomarkers associated with adverse effects on earthworm physiology and ecology, filling existing knowledge gaps in understanding mechanisms of toxicity. We introduce a conceptual framework linking pesticide classes to specific biochemical markers and ecological consequences, highlighting how certain pesticide groups (such as neonicotinoids and organophosphates) pose significant risks. Our synthesis reveals that sub-lethal effects, including enzyme activity disruptions, DNA damage, and reproductive impairments, are crucial indicators of environmental health. These findings advance current understanding by pinpointing sensitive biomarkers and emphasizing the importance of sustainable pesticide management strategies to mitigate ecological harm and protect terrestrial ecosystems.
 
Novelty Statement | This study presents a comprehensive synthesis of recent research on the effects of various pesticides on earthworm health, highlighting novel insights into the mechanisms of toxicity and ecological implications that have not been systematically reviewed before. 

Introduction

The widespread use of synthetic pesticides has greatly increased the agricultural productivity by reducing losses resulted from presence of weeds, insects, and diseases ; ; . However, this intensification has led to declining biodiversity in farm ecosystems ; . Pesticide droplets often settle on both pests and non-target organisms, affecting a broad range of species sharing similar traits . Despite efforts to limit their spread, pesticides drift, volatilize, runoff, and penetrate soil, impacting surrounding environments ; , b; . These pesticides (including insecticides, herbicides, fungicides, and others) target specific pests but can harm soil and environmental life at even trace levels, posing ecological and health risks ; ; ; -c; ; . Most pesticides contaminate air, water, and soil, with less than 0.1% reaching intended targets, leading to accumulation in ecosystems and entering the food chain. Recognizing soil’s vital ecological role (supporting food production, nutrient cycling, carbon storage, and biodiversity) is increasingly important. Yet, agricultural and industrial activities intensify pressure on this finite resource, especially as pesticides contaminate soil directly or via runoff ; -e; . Earthworms, key soil inhabitants, are particularly vulnerable they absorb pesticides through skin contact and ingestion, depending on chemical adsorption and soil conditions -b; ; ; , b). Moving up to 6.3 tons of soil per hectare annually, earthworms process organic and mineral matter, influencing soil health profoundly (Mosleh et al., 2003a, b). Pesticides can harm earthworms by reducing cocoon production, juvenile density, and growth, and increasing mortality and tissue damage; they also induce genetic, biochemical, and physiological stress (Zhang et al., 2014a). Despite developing some defenses, earthworms remain sensitive bioindicators of soil contamination ideal for ecotoxicological assessments due to their ease of study and consistent responses . Their burrowing and casting activities enhance soil structure, nutrient cycling, and plant growth, making them vital for sustainable agroecosystems . As key ecosystem engineers, earthworms support soil fertility and help process pollutants, with their abundance reflecting soil health and contamination levels . Consequently, they are frequently used to evaluate the ecotoxic effects of pesticides, highlighting the importance of understanding and mitigating chemical impacts on soil biodiversity -e).

Methodology

The methodology for assessing the impact of pesticides on earthworm health in a comprehensive review involves a systematic approach to gather, evaluate, and synthesize existing research. Initially, a thorough literature search is conducted across multiple scientific databases such as PubMed, Scopus, Web of Science, and Google Scholar to identify relevant studies spanning all years. Inclusion criteria are established to select peer-reviewed articles, experimental studies, field surveys, and meta-analyses that investigate various aspects of pesticide exposure and earthworm health. Data extraction involves collecting information on pesticide types, concentrations, application methods, duration of exposure, earthworm species studied, and health parameters assessed (e.g., survival rate, reproduction, behavioral changes, physiological and biochemical markers). The review critically analyzes experimental designs, methodologies, and results to identify patterns, correlations, and disparities across different studies. Additionally, the methodology may include a quality assessment of the selected studies to ensure reliability and validity. Synthesis of data involves qualitative and quantitative approaches, such as meta-analysis if applicable, to derive overarching conclusions about the effects of pesticides on earthworms. Overall, this methodology aims to provide a comprehensive understanding of how pesticides influence earthworm health over time and across various environmental contexts. While numerous reviews have addressed the general effects of pesticides on soil organisms, many lack a focused synthesis on earthworms, particularly in relation to emerging pesticide formulations and application methods. This review uniquely consolidates recent data on the sub-lethal and long-term impacts of various pesticide classes on earthworm physiology, behavior, and reproductive success, highlighting inconsistencies and gaps in current methodologies.

Pesticide production and consumption in the world

In 2022, global pesticide active ingredient use reached 3.7 million tons up 4% from 2021, 13% over the past decade, and three times more than in 1990. Compared to the 1990s, pesticide use has increased by 48% for insecticides, 54% for fungicides and bactericides, and 121% for herbicides. During this period, herbicides’ share of total pesticide use grew from 40–50%, while insecticides decreased from 26% to 22%, and fungicides and bactericides from 25% to 22% (see Figure 1). In 2022, pesticide exports totaled approximately 6.9 million tonnes (down 1% from 2021) but their value rose by 13% to USD 48.8 billion. Between 2013 and 2022, exports increased by 50%, and their value grew by 53%. The top five pesticide-consuming countries, from most to least, are China, the United States, Argentina, Thailand, and Brazil (FAO, 2024; .

Ecological classification of earthworms

Over 7,000 earthworm species have been described globally, spread among 23 families and 700 genera. There are three categories into which earthworms fall depends on the area of soil that earthworms live in, as well as their feeding and burrowing habits. Lumbricus castaneus is an example of an epigeic species, which live and feed on soil surface. Anecic earthworms, such as Lumbricus terrestris, may emerge from their permanent vertical burrows in soil to feed onto surface litter. Endogeic species, such as Allolobophora chlorotica, feed on soil and dwell in transient horizontal burrows . Each category uniquely supports ecosystem services, with epigeic focusing on surface processes, anecic on vertical mixing and aeration, and endogeic on subsurface soil health. Together, they play vital roles in maintaining soil fertility and ecosystem productivity.

LC50 of pesticides on earthworms

Eisenia fetida

Tables 1 and 2, the LC50 values of different pesticides on Eisenia foetida are shown using filter paper contact technique and soil mixing technique. E. fetida exposed to atrazine (0.1–5 µg/cm²) had a mean 96-h LC50 of 2.9 µg/cm². For cyanazine (0.5–10 µg/cm²), the LC50 was 4.9 µg/cm², and for chlorpyrifos (1–15 µg/cm²), it was slightly higher at 8.3 µg/cm² . LC50 value of chlorpyrifos was 0.047 and 0.037 µg/cm2 at 24 and 48 hours by Rao et al. (2003). The LC50 of azodrin was 0.46 µg/cm² via paper contact after 48 hours, and 132 mg/kg in artificial soil after 14-d . Lindane was more toxic than deltamethrin to E. fetida over 14-d, with LC50 values of 162.1 mg/kg and 432.9 mg/kg soil, respectively . The LC50 values for benomyl, carbendazim and lambda-cyhalthrin were 633, >1000, and 23.9 mg/kg into tropical artificial soil, 22, 5.8, and 99.8 mg/kg into artificial soil, 14.6, 4.1, and 139.9 mg/kg into loamy sand soil, respectively . The LC50 values for profenofhos were 4.56 and 3.55 µg/cm2 after 24 and 48 hours, respectively . Wang et al. (2012) found that insect growth regulators were moderately toxic to worms (LC50: 117.6–564.6 µg/cm² at 48 hours), pyrethroids were highly toxic (LC50: 10.55–25.7 µg/cm²), and neonicotinoids were extremely toxic (LC50: 0.0088–0.45 µg/cm²). Using an artificial soil bioassay, neonicotinoids exhibited greater acute toxicity than antibiotics, carbamates, insect growth regulators, organophosphates, and pyrethroids, which were the least hazardous. Terbuthylazine herbicide was the most ecologically harmful to E. fetida, followed by galition (fenitrothion and malathion) insecticides, and metaldehyde limacid . The 96-h acute toxicity test in natural soil showed that carbendazim was more toxic than captan . The LC50 of acetamiprid, imidacloprid, nitenpyram, thiacloprid and clothianidin was 2.69, 3.05, 4.34, 2.68 and 0.93 mg/kg artificial soil, respectively , b). It was found that carbendazim is highly toxic to E. fetida, with a 14-day LC50 of 2 mg/kg. Dimethoate shows moderate toxicity (LC50 of 28 mg/kg), whereas trichlorfon, tebuconazole, and prochloraz exhibit relatively low toxicity . According to Chen et al. (2018), tribenuron-methyl exhibited low toxicity to E. fetida in studies using contact filter paper and artificial soil. Isoprocarb increased mortality in a dose-dependent manner, with LC50 values of 8.2 μg/cm² (48-h) on filter paper and 3.37 mg/kg (14-d) in soil . In the artificial soil, the 14-day LC50 value of flupyradifurone was 157.6502 mg/kg for juvenile E. fetida and 186.9773 mg/kg to adult E. fetida . Acetamiprid was 167.9-times more toxic for E. fetida than tetraconazo after a 7-day break. Likewise, acetamiprid’s toxicity was 142.2 times greater than tetraconazo’s over a 14-d interval .

Table 1: LC50 of pesticides to Eisenia foetida by filter paper contact (µg/cm2) technique.

Pesticide

Time (h)

LC50

Ref.

Chlorpyrifos, Atrazine, Cyanazine

96

8.3, 2.9, 4.9

Lydy and Linck, 2003

Chlorpyrifos

24 and 48

0.047 and 0.037

Rao et al., 2003

Azodrin

48

0.46

Rao and Kavitha, 2004

Profenofos

24 and 48

4.56 and 3.55

Reddy and Rao, 2008

Acetamiprid, Clothianidin, Imidacloprid, Nitenpyram, Thiacloprid, Abamectin, Emamectin benzoate, Ivermectin, Buprofezin, Chlorfluazuron, Hexaflumuron, Tebufenozide, Cyhalothrin, Cypermethrin, Fenpropathrin, Lambda-cyhalthrin, Carbosulfan, Isoprocarb, Metolcarb, Promecarb, Chlorpyrifos, Phoxim, Pyridaphenthion, Triazophos

48

0.0088, 0.28, 0.027, 0.22, 0.45, 23.08, 30.2, 4.40, 564.6, 536.2, 117.6, 508.1, 24.30, 10.63, 10.55, 25.7, 75.75, 3.64, 9.18, 10.58, 14.19, 54.65, 3.84, 14.21

Wang et al., 2012

Tribenuron-methyl, Tebuconazole

24 and 48

377.9 and 9.4 –135.6 and 5.7

Chen et al., 2018

Isoprocarb

24 and 48

32.04 and 8.20

Gu et al., 2021

Table 2: LC50 of pesticides to Eisenia foetida by soil mixing (mg/kg soil) technique.

Pesticide

Soil type

Time (d)

LC50

Ref.

Azodrin

Artificial

7 and 14

171 and 132

Rao and Kavitha, 2004

lindane, deltamethrin

Artificial

14

162.1, 432.9

Shi et al., 2007

Benomyl, Carbendazim, Lambda-cyhalthrin

Tropical artificial

2

633, >1000, 23.9

Garcia et al., 2008

Artificial

22.0, 5.8, 99.8

Loamy sand

14.6, 4.1, 139.9

Acetamiprid, Clothianidin, Imidacloprid, Nitenpyram, Thiacloprid, Abamectin, Emamectin benzoate, Ivermectin, Buprofezin, Chlorfluazuron, Hexaflumuron, Tebufenozide, Cyhalothrin, Cypermethrin, Fenpropathrin, Lambda-cyhalthrin, Carbosulfan, Isoprocarb, Metolcarb, Promecarb, Chlorpyrifos, Phoxim, Pyridaphenthion, Triazophos

Artificial

7 and 14

1.72 and 1.52 –7.44 and 6.06, 3.15 and 2.82, 4.42 and 3.91, 12.13 and 10.96, 31.50 and 27.86, 196.2 and 175.3, 68.11 and 56.34, 425.1 and 363.3, 406.5 and 381.8, 420.1 and 374.2, 434.8 and 386.7, 1530 and 1369, 1467 and 1272, 1532 and 1246, 1623 and 1370, 146.8 and 130.1, 69.4 and 60.8, 108.1 and 93.8, 31.23 and 28.43, 421.3 and 384.9, 1083 and 901.5, 273.3 and 243.7, 381.4 and 347.5

Wang et al., 2012

Galition, Fenitrothion+Malathion, Terbuthylazine, Metaldehyde

Artificial

14

368.25, 1.264, 3.66

Jovana et al., 2014

Carbendazim, Captan

Natural

4

5.38, 10.41

Mandal et al., 2015

Imidacloprid, Acetamiprid, Nitenpyram, Clothianidin, Thiacloprid

Artificial

14

3.05, 2.69, 4.34, 0.93, 2.68

Wang et al., 2015a, b

Trichlorfon, Dimethoate, Carbendazim, Tebuconazole, Prochloraz

Artificial

14

122, 28, 2, 180, 261

Rico et al., 2016

Tribenuron-methyl, Tebuconazole

Artificial

7 and 14

1007.3 and 746.3, 511.0 and 287.9

Chen et al., 2018

Isoprocarb

Artificial

7 and 14

6.95 and 3.37

Gu et al., 2021

Flupyradifurone

Artificial

14

186.9773 (adult) and 157.6502 (juveniles)

Qiao et al., 2022

Acetamiprid, Tetraconazo

Artificial

7 and 14

1.96 and 1.54, 329 and 219

An et al., 2025

Eisenia andrei

LC50 values of various pesticides against E. andrei under different exposure regimes are shown in Table 3. The LC50 values were 492-154, 23.4-13.5, and 7.10-38.9 mg/kg in artificial soil, while they were 481-106, 8.46-11.9, and 2.70-41.5 mg/kg in loamy sand soil for chlorpyrifos, carbofuran, and carbendazim at 20 and 26°C, respectively . The toxicity for mixture of chlorpyrifos, and cypermethrin (LC50= 35.06) was significantly higher than either of cypermethrin (LC50 = 86.04), and chlorpyrifos (LC50 = 116) individually onto Eisenia fetida Andrei . The LC50 value is 25.34 mg/kg at 14-day for organophosphate azinphos-methyl on E. Andrei . Quantified impact of imidacloprid, thiametoxam, fipronil, and captan (thiram + carboxin) on reproduction, survival, and behavior of E. andrei, with exception of imidacloprid (LC50 = 53.25 mg/kg soil), none of pesticides tested caused mortality . LC50 values were 102.035, 10.149, 249.804, and 558.376 mg/kg dry soil after 1-d of treatment for thiacloprid, esfenvalerate, dimethenamid-p, and prosulfocarb, respectively .

Aporrectodea caliginosa

Mosleh et al. (2003a) studied toxicity of cypermethrin, profenofos, aldicarb, atrazine, metalaxyl and chlorfluazuron to earthworms, A. caligenosa. The result was that aldicarb (LC50 = 0.68) was the most toxic to A. caligenosa in artificial soil after 28 days, followed in order by cypermethrin (LC50 = 72.96), profenofos (LC50 = 127), chlorfluazuron (LC50 = 139.9), atrazine (LC50 = 381.2), and metalaxyl (LC50 = 518) (Table 4). The LC50 value of acetamiprid was 0.199 and 0.009 after 5 and 10 day for A. caliginosa in alluvial soil . The LC50 values were (334.27 and 55.45 mg/kg) into clay soil, and (415.9 and 25 mg/kg) into sandy clay loam soil for fenitrothion after 5 and 10 days of exposure. LC50 values were (0.93 and 0.41 mg/kg) into clay soil, and (55.28 and 10.65 mg/kg) into sandy clay loam soil after 5 and 10 day of exposure for thiobencarb (El-Aswad et al., 2023a-b). Atrazine had higher intrinsic toxicity than metribuzin to earthworms (A. caligenosa), with LC50 of 1.693 and 0.026 μg/cm2 after 48 and 72 hours by filter paper contact test. LC50 was reduced from 11.121 to 3.118 and 164.824 to 19.113 mg/kg into clay soil, from 32.221 to 17.33 and 324.141 to 41.028 mg/kg into clay soil: Sandy clay loam soil (1:1), and from 41.234 to 30.804 and 462.255 to 70.902 mg/kg into sandy clay loam soil of atrazine, and metribuzin after 5 and 10 days .

Table 3: LC50 of pesticides to Eisenia andrei, in different exposure techniques.

Pesticides

Exposure method

Time (d)

LC50

Ref.

Chlorpyrifos, Carbofuran, Carbendazim

In artificial soil (mg/kg dry soil) at 20 and 26 °C

28

492 and 154, 23.4 and 13.5, 7.10 and 38.9

De Silva et al., 2009

In loamy sand soil (mg/kg dry soil) at 20 and 26 °C

481 and 106, 8.46 and 11.9, 2.70 and 41.5

Cypermethrin, Chlorpyrifos, Cypermethrin+Chlorpyrifos

In artificial soil (mg/kg dry soil)

14

86.04, 116.00, 35.06

Zhou et al., 2011

Azinphos-methyl

In artificial soil (mg/kg dry soil)

14

25.34

Jordaan et al., 2012

Imidacloprid, Thiametoxam, Fipronil, Carboxin+Thiram

In artificial soil (mg/kg dry soil)

14

25.53, >1000, >1000, >1000

Alves et al., 2013

Difenoconazole+Fludioxonil, Fluopyram+Tebuconazole, Pyrimethanil, Thiram+Carboxin, Flutriafol+Thiabendazole, Fluroxypyr-meptyl, Thiamethoxam

On filter paper (µg/cm2)

1, 2 and 3

1.4, 0.6 and 0.3, 5.9, 2.9 and 1.0, 40.9, 6.5 and 2.7, 31.8, 8.2 and 1.1, 74.3, 16.2 and1.2, 19.7, 17.3 and 14.1, 724.0, 110.3 and 19.2

Velki et al., 2019

Thiacloprid, Esfenvalerate, Dimethenamid-p, Prosulfocarb

In artificial soil (mg/kg dry soil)

2

102.035, 10.149, 249.804, 558.376

Lackmann et al., 2023

Lumbricus terrestris

L. terrestris were exposed for commercial formulations of aldicarb and endosulfan at 2, 7, 15 days, with LC50 were 12.29, 5.82 and 3.36 mg/kg for endosulfan, 3.09, 1.51 and 1.5 mg/kg for aldicarb (Mosleh et al., 2003b). The LC50 value was 26.804 and 7.001 mg/kg on earthworms in sandy loam soil after 21 and 42 days of glyphosate exposure (Table 4) .

Effect of pesticides on enzyme activities in earthworms

Acetylcholinesterase

Carbamate and organophosphorus pesticides primarily inhibit acetylcholinesterase (AChE). Using a standardized paper contact method, E. fetida shows a time-dependent decrease in AChE activity when exposed to azodrin and chlorpyrifos . As chlorpyrifos and imidacloprid concentrations increased, AChE activity in E. foetida was progressively suppressed . Azodrine also decreased AChE activity in a concentration-dependent manner . Diazinon reduced AChE activity in E. fetida body wall and gut , with the lowest activity observed at 120 ng/cm² of temephos after 1 and 2 hours . Methiocarb significantly decreased AChE activity , while ethyl-4-chlorophenyl, ethyl-4-bromophenyl carbamates, and propoxur caused weak inhibition . Isoprocarb also decreased AChE activity , and compounds like carbendazim, trichlorfon, dimethoate, prochloraz, and tebuconazole significantly inhibited AChE in artificial soil . Exposure to imidacloprid over 3 to 21 days reduced AChE activity in E. fetida by up to 46.7%, with inhibition rates of 28.9–46.7% . Flupyradifurone at 25 and 50 mg/kg maintained AChE inhibition over 28 days . Conversely, atrazine exposure increased AChE activity in E. fetida . In E. andrei, carbaryl primarily inhibits AChE competitively . Organophosphates like diazinon and chlorpyrifos suppressed AChE by 72–87% within a day and remained suppressed in A. caliginosa for 14-d . Parathion caused rapid AChE inhibition (~70%) within three days, reaching 80–90% after a week; similar patterns were observed in A. chlorotica, though at a slower pace . AChE activity in A. caliginosa and A. caligenosa was inhibited after exposure to lambda-cyhalothrin, chlorpyrifos-ethyl, and fungicides like myclobutanil and folpet within three days ; . L. terrestris showed significant AChE inhibition as an indirect measure of chlorpyrifos bioavailability . Conversely, Dendrobaena veneta exhibited increased AChE activity after a week of glyphosate exposure at 2.16 mg/kg soil . Chlorpyrifos and cypermethrin suppressed AChE in Eudrilus eugeniae in a dose and region-dependent manner, while glyphosate had no significant effect on AChE activity in E. eugeniae, Libyodrilus violaceus, and Alma millsoni . Variations in AChE response are likely influenced by pesticide type and concentration, earthworm species, and soil properties .

Glutathione-S-transferase

When glutathione is used, glutathione-S-transferase (GST) plays a vital role in detoxifying electrophilic chemicals . Elevated GST levels can enhance defense against pesticides, making GST a potential biomarker for pollution monitoring . Studies show that earthworms exposed to chemicals like glyphosate, QYR301, aldrin, endosulfan, and lindane exhibit significantly increased GST activity . Similarly, GST activity in

Table 4: LC50 of pesticides to Aporrectodea caliginosa and Lumbricus terrestris in different exposure systems.

Earthworms

Pesticides

Exposure method

Exposure time

LC50

Ref.

A. caliginosa

Aldicarb, Cypermethrin, Profenofos, Chlorfluazuron, Atrazine, Metalaxyl

In artificial soil (mg/kg dry soil)

28 days

0.68, 72.96, 127.00, 139.90, 381.20, 518.00

Mosleh et al., 2003a

Acetamiprid

In alluvial soil (µg/g soil)

5 and 10 days

0.199 and 0.009

Abdel-Raheem et al., 2023

Fenitrothion, Thiobencarb

On filter paper (µg/cm2)

48 and 72 hours

83.16 and 24.67, 288.26 and 39.98

El-Aswad et al., 2023a

In clay soil (µg/g soil)

5 and 10 days

334.27 and 55.45, 0.93 and 0.41

In sandy clay loam soil (µg/g soil)

415.90 and 25.00, 55.28 and 10.65

Atrazine, Metribuzin

On filter paper (µg/cm2)

24, 48 and 72 hours

43.007, 10.260 and 0.910, 65.620, 1.693 and 0.063

Fouad et al., 2023b

In clay soil (µg/g soil)

5 and 10 days

11.121 and 3.118, 164.824 and 19.113

In sandy clay loam soil (µg/g soil)

41.234 and 30.408, 462.255 and 70.902

In a mixture of clay soil and sandy clay loam soil (µg/g soil)

32.221 and 17.330, 324.141 and 41.028

L. terrestris

Endosulfan, Aldicarb

In artificial soil (mg/kg dry soil)

2, 7 and 15 days

12.29, 5.82 and 3.36, 3.09, 1.51 and 1.50

Mosleh et al., 2003b

Glyphosate

In sandy loam soil (g/m2)

21 and 42 days

26.804 and 7.001

Stellin et al., 2018

E. fetida rises upon exposure to compounds such as flupyradifurone, azoxystrobin, fomesafen, sulfentrazone, and pyraclostrobin ; Zhang et al., 2014b; ; . Long-term exposure to cyflumetofen also activates GST . Conversely, some chemicals like cyantraniliprole initially decrease and later increase GST activity, while thifluzamide inhibits it ; . Soil contamination with oxyfluorfen causes significant GST reductions in earthworms such as Allolobophora molleri, E. fetida, and L. terrestris . Similarly, exposure to lambda-cyhalothrin, chlorpyrifos-ethyl, and other pesticides over 14-d suppresses GST activity in A. caliginosa nocturnus, especially at higher concentrations .

Superoxide dismutase

Superoxide dismutase (SOD) is the primary antioxidant enzyme defending against reactive oxygen species. Owagboriaye et al. (2020) found that glyphosate-treated earthworms (L. violaceus, E. eugeniae, A. millsoni) exhibited significantly higher SOD activity than unexposed ones. Conversely, Liu et al. (2020) observed different SOD responses in E. fetida after trifloxystrobin exposure. Other studies reported significant variations in SOD activity in E. eugeniae and E. fetida when exposed to nitenpiram, malathion, triflumezopyrim, and sulfoxaflor -b; ; ; . Exposure to azoxystrobin, QYR30, and flupyradifurone increased SOD activity in E. fetida, indicating detoxification and oxidative stress mitigation . SOD activity also rose with higher sulfentrazone levels, showing a dose-dependent effect . A maximum dose of spirotetramat (2.5 mg/kg) significantly elevated SOD activity during exposure . Conversely, atrazine and thifluzamide inhibited SOD activity, while increasing concentrations of isoprocarb reduced it ; . Responses to cyantraniliprole varied, with initial decreases followed by increases in SOD activity .

Catalase

Catalase (CAT) converts hydrogen peroxide into oxygen and water, reducing its toxicity . In E. fetida, CAT activity decreases at higher chlorpyrifos doses but remains unaffected at low levels . Tebuconazole (0.5, 5, 50 mg/kg soil) showed no significant impact on CAT activity , and azoxystrobin did not increase CAT activity . Exposure to atrazine and isoprocarb reduced CAT activity, while diazinon increased it ; . Triflumisopyrim initially elevated CAT activity, which normalized after 28 days . Similarly, flupyradifurone increased CAT activity . Some fungicides, like spirotetramat, pyraclostrobin, thifluzamide, and QYR301, initially activated CAT but inhibited it over time ; Yao et al., 2020). Cyantraniliprole caused an initial decrease followed by an increase in CAT activity . During 7-21 days, sulfentrazone increased CAT activity at all concentrations, while at 7 and 28 days, there was no significant change . In A. caligenosa nocturnus, seven days of exposure to lambda-cyhalothrin, chlorpyrifos-ethyl, folpet, fosetyl-Al, myclobutanil, and metalaxyl decreased CAT activity .

Cellulase

Cellulase is a digestive enzyme involved in cellulose breakdown. Acute deltamethrin exposure decreased E. fetida cellulase activity, while lindane increased it . Fomesafen at 500 mg/kg for 14 days markedly inhibited cellulase activity in E. fetida (Zhang et al., 2014b), and high-dose spirotetramat also significantly reduced activity during exposure . Sublethal doses of dichlorovos and endosulfan significantly inhibited cellulase activity in E. foetida . Exposure to imidacloprid and acetamiprid increased cellulase activity, peaking at 14 days before returning to normal. Combined chlorpyrifos with these insecticides caused initial activation after one day, with levels normalizing after seven days . Patnaik and Dash (1993) reported reduced cellulose activity in three earthworm species (Drawida willsi, Lampito mauritii, and Octochaetona surensis) after malathion exposure.

Effect of pesticides on histology in earthworms

Cellular and tissue damage in earthworms exposed to hazardous pesticides has been documented through histopathological studies. Imidacloprid induces dose and time dependent genotoxicity in coelomocytes of Metaphire posthuma and E. eugeniae, with midgut cells showing abnormal nuclei, reduced cytoplasm, and cellular breakdown as exposure increases ; . Multiple neonicotinoids (including thiamethoxam, dinotefuran, acetamiprid, clothianidin, nitenpyram, and thiacloprid) damage midgut and epidermal tissues ; , with acetamiprid causing severe structural disintegration . Acetamiprid also induces alterations in epidermal, muscle, and vascular tissues of Aporrectodea giardi . Other pesticides, such as abamectin, cause posterior segment loss, fragmentation, and body thinning in E. andrei across all tested concentrations . Lindane exposure results in vacuolation and muscle-epidermis separation, effects that intensify with higher doses of dimethoate and chlorpyrifos, leading to cellular destruction in A. caliginosa ; ; . Dimethoate affects neurosecretory cell morphology and urosecretory processes in Eudichogaste Kinneari . Profenofos treatment causes body rupture, bleeding, glandular proliferation, and muscle disintegration in E. fetida . Exposure to triazophos and deltamethrin damages the post-clitellar region of E. eugeniae, causing tissue rupture and necrosis . Monocrotophos induces rupture, necrosis, and tissue fusion in earthworm body walls, while also negatively impacting intestinal histology and microbial populations in L. mauritii ; . High concentrations of monocrotophos and glyphosate cause lesions, skin undulation, and deformities in E. eugeniae, with glyphosate damaging intestinal tissue in Pheretima elongata ; . Glyphosate exposure leads to cuticular degeneration, cytolysis, vacuolization, and muscle damage in Nsukkadrilus mbae . Similarly, atrazine causes vacuolation, loss of nuclei, and tissue disorganization in N. mbae . Exposure to butachlor results in vacuolation and glandular enlargement in Perionyx sansibaricus and E. fetida ; . Sulfentrazone damages the digestive and epidermal tissues of E. fetida after 21 days, with cellular disintegration observed . Triazole fungicides cause destruction of epidermal cells and disorganization of muscle layers, with higher concentrations inducing cytoplasmic staining and cell death .

Genotoxic effects of pesticides on earthworms

E. fetida exhibits antioxidant responses to lipid peroxidation and DNA damage caused by subchronic exposure to acetamiprid and imidacloprid . Imidacloprid at 0.1–2 mg/kg soil can induce DNA damage after 21 days, as shown by sperm deformity assessments . Nitenpyram also causes notable DNA damage in E. fetida, with effects increasing dose-dependently . DNA damage metrics, such as tail moment, rise with higher cyantraniliprole concentrations and longer exposure . Endosulfan and triflumezopyrim similarly induce DNA damage , while deltamethrin + triazophos and profenofos + cypermethrin show moderate genotoxicity . Chlorpyriphos increases DNA damage and reactive oxygen species levels, leading to oxidative stress at low concentrations ; . Glyphosate formulations are cytotoxic and genotoxic both in vivo and ex vivo , but glyphosate alone shows no clear DNA harm risk in Pheretima peguana . Conversely, paraquat is clastogenic and aneugenic , and atrazine damages E. foetida DNA . Sulfentrazone and QYR301 induce oxidative stress, lipid peroxidation, and DNA damage in E. fetida . Short-term exposure to diuron (0–5 mg/kg) causes minimal oxidative stress and DNA damage , while chloridazon, oxadiazon, bentazone, and pendimethalin increase DNA damage and chromosomal abnormalities in Eisenia hortensis ; . Azostrobin triggers oxidative stress and DNA damage in E. fetida , and carbendazim and metalaxyl-M cause significant DNA damage in E. foetida coelomocytes, with effects related to dose and exposure duration ; .

Effects of pesticides on growth of earthworms

Although glyphosate is relatively non-toxic to Aporrectodea trapezoides, Aporrectodea rosea, A. caliginosa, and Aporrectodea longa in field studies, it has been shown to reduce A. caliginosa growth in laboratory settings, decrease cocoon viability, and lower juvenile counts in the field. It also affects reproduction and development in E. foetida , leading to fewer juveniles. Similarly, glyphosate decreases cocoon viability and juvenile production in E. fetida . Laboratory tests reveal that 2,4-D and glyphosate significantly impact E. foetida reproduction and development, with treated soils lacking cocoons and juveniles . These chemicals, along with carbendazim, dimethoate, and copper oxychloride, cause dose-dependent reductions in E. fetida growth . Both glyphosate and chlorpyrifos reduce E. fetida andrei feeding activity in field and lab conditions, though low chlorpyrifos concentrations do not affect reproduction . The weight of E. fetida is more sensitive than mortality to methamidophos and acetochlor exposure . Increased butachlor concentrations decrease E. fetida biomass and cocoon formation , and inhibit P. sansibaricus growth and cocoon production . Lindane exposure reduces the biomass of E. eugeniae . Pesticides such as chlorpyrifos, diazinon, aldicarb, profenofos, cypermethrin, chlorfluazuron, metalaxyl, and atrazine slow the growth of A. caliginosa ; , with diazinon and chlorpyrifos notably reducing growth during exposure . Endosulfan significantly decreases juvenile A. trapezoides weight within 35-days when applied at normal rates, while fenamiphos has similar effects only in the field. Malathion reduces E. foetida weight and sperm viability, with parathion also impairing E. andrei growth . Neonicotinoids such as imidacloprid, nitenpyram, acetamiprid, thiacloprid, and clothianidin are highly toxic to E. fetida, significantly inhibiting fecundity , b), with high doses adversely affecting reproduction, growth, and physiology of M. posthuma and E. eugeniae . Acetamiprid induces mortality in A. giardi proportional to dose and exposure time . Elevated abamectin levels decrease E. andrei reproductive efficiency , and cyantraniliprole exposure dramatically reduces E. fetida weights, likely due to loss of proteins, fats, and glycogen .

Effect of pesticides on the behavior of earthworms

These sublethal, organism-wide effects can occur even at low pesticide exposures, making behavioral changes highly sensitive and ecologically relevant . Earthworms play a crucial role in soil structure and aeration; their soil preference or avoidance can indicate soil quality issues . Avoidance behavior is a well-established, sensitive indicator of soil pesticide contamination, as pesticides have been shown to influence earthworm avoidance responses. For instance, exposure to sulcotrione and penoxsulam affected avoidance in standardized soils , while 2.5 and 5 mg/kg esfenvalerate increased avoidance . Similarly, E. fetida and E. andrei avoided glyphosate-treated soils , though low chlorpyrifos concentrations showed no effect. Elevated doses of deltamethrin + triazophos or profenofos + cypermethrin altered earthworm behavior, causing body folding and reduced activity . Additionally, imidacloprid significantly affected burrow depth, length, and branching in Aporrectodea nocturna and Allolobophora icterica.

Conclusion

This review underscores the complex and multifaceted threats that pesticides pose to earthworm populations and, consequently, to soil health and ecosystem stability. Analysis of the existing literature reveals several consistent patterns: organophosphates and carbamates emerge as the most acutely toxic pesticide classes, exhibiting high mortality rates in earthworms at recommended agricultural doses, while certain neonicotinoids, though less immediately lethal, induce sub-lethal effects such as impaired reproduction and growth over time. Among various biomarkers, reductions in reproductive output, alterations in antioxidant enzyme activities (such as catalase and superoxide dismutase), and increased DNA damage measured via comet assays are the most sensitive indicators of pesticide-induced stress, signaling early and sustained biological impacts before mortality occurs. Pesticides can also induce significant histopathological alterations, including degeneration of the epidermis and gut epithelium, alongside genotoxic effects that compromise cellular integrity and genetic stability, potentially impairing earthworm populations and their ecological functions. To address these issues, policymakers should implement strict regulations to limit or ban the use of the most toxic pesticide classes, promote integrated pest management strategies that prioritize biological control and biopesticides, mandate regular environmental monitoring using sensitive biomarkers for early detection, and educate farmers and stakeholders about the ecological costs of pesticide overuse and sustainable soil management practices. Future research should focus on longitudinal field studies to elucidate chronic and cumulative effects, standardize biomarker protocols for cross-study comparison, investigate potential bioaccumulation and transfer of pesticides through soil food webs, and develop earthworm-based bioassays as rapid, cost-effective soil health assessment tools. By synthesizing these patterns and translating them into targeted actions and research priorities, we can better safeguard earthworm populations, preserve vital ecosystem services, and promote sustainable agricultural practices for future generations.

Declarations

Funding

Not applicable.

IRB approval

The review was approved by the Department of Chemistry and Pesticide Technology.

Ethical statement

The study is a literature review on pesticides’ effects on earthworms, involving no direct experimentation. It uses publicly available, peer-reviewed sources, properly cites all data, and adheres to ethical standards. No ethical approval was needed since it does not involve new experiments or fieldwork.

Consent for publication

The review contains no such material that may be unlawful, defamatory, or which would, if published, in any way whatsoever, violate the terms and conditions as laid down in the agreement.

Generative AI and AI-assisted technology statement

No Generative AI and AI-assisted technologies wer used in the writing process.

Conflict of interest

The author has declared no conflict of interest.

Figures

Figure 1:
Figure 1:.

Worldwide pesticides consumption.

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Assessing the Impact of Pesticides on Earthworm Health: A Comprehensive Review · ResearchersLinks