Research Article

Rearing of Galleria mellonella Under Controlled Environments: A Study on Diets, Temperature, and Management Strategies

Hussain Ali1*, Mirzakhmedov Muhammadjon Fazliddin ogli2, Saparov Kuanishbay Abdirakhmanovich3, Sultanov Esbosin Kosimbetovich3 and Embergenov Muratbay Amanbaevich4,5

1Directorate of Agriculture Research Peshawar, Khyber Pakhtunkhwa, Pakistan; 2Namangan State University, Namangan, Uzbekistan; 3Nukus State Pedagogical Institute Named after Ajiniyaz, Nukus, Uzbekistan; 4Republican Scientific and Production Centre for Termite Control at the Institute of Zoology, Academy of Sciences of Uzbekistan, Tashkent, Uzbekistan; 5Institute of Zoology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan.

Received | July 27, 2025; Accepted | October 28, 2025; Published | November 27, 2025

*Correspondence | Hussain Ali, Directorate of Agriculture Research Peshawar, Khyber Pakhtunkhwa, Pakistan; Email: [email protected]

Citation | Ali, H., M.M.F. Ogli, S.K. Abdirakhmanovich, S.E. Kosimbetovich and E.M. Amanbaevich. 2025. Rearing of Galleria mellonella under controlled environments: a study on diets, temperature, and management strategies. Pakistan Journal of Agricultural Research, 38(4): 33-38.

DOI | https://dx.doi.org/10.17582/journal.pjar/2025/38.4.33.38

Keywords | Galleria mellonella, wax, Development, Instar, Mortality, Neem

Copyright: 2025 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Honey beekeeping is practiced in Khyber Pakhtunkhwa on large scale. Two honey bee species are reared for honey production, the Asian honey bee A. cerana) and the exotic honey bees A. mellifera. Both have good potential for honey production and more 0.5 million people of Khyber Pakhtunkhwa are involved in this sector. The sector is greatly challenged by climate change, floral resources, agricultural pesticides application, pests and diseases. Among the pests mites and wax moth have great impact on honey beekeeping (Ali et al., 2022).

Wax moths tend to cause more problems in leftover combs and weak colonies. Their larvae typically eat wax combs, pollen, larval exuviae, and honey (Graham, 1992). The presence of wax moth infestation can be detected by silken threads or tunnels within the wax combs (Hepburn and Hepburn, 2005). Heavy larval feeding can break down the combs into piles of debris made up of frass and wax moth webbings, and the larvae may even bore into the midribs (bases) of the combs. They usually prefer dark combs, which are those used for brood rearing, making these combs more vulnerable to damage (Ellis et al., 2013).

The G. mellonella consume wax combs, discarded honey bee larval skins, honey and pollen (Graham, 1992). Their occurence within frames is indicated by the formation of silken threads, which often create tunnel-like structures inside the wax. This pest tends to cause more damage in weak or stressed colonies compared to stronger ones (Ellis et al., 2013; Sohail et al., 2017; Hood et al., 2003).

Average temperatures ranges from 25-30°C, to complete the developmental stages the duration is 5 days for the egg hatching, 30-40 days larva, 14 days for pupal stage. Under warmer conditions (33.8 °C), the larvae go through 8 to 9 molts during their development (Charriere and Imdorf, 1999).

Chemical fumigants are commonly used to control greater wax moth (GWM) in beekeeping. These include ethylene bromide, methyl bromide, sulphur, phosphine etc. However, these fumigants are toxic to bees and beneficial insects (Ellis et al., 2013).

The present study was conducted to evaluate the larval development of the greater wax moth (Galleria mellonella) on various diets across different temperature conditions, and to assess the effectiveness of multiple control methods for its management under laboratory conditions.

Materials and Methods

Location

The trials were carried out at Entomology Research Programme at Agricultural Research Institute (ARI), Tarnab farm Peshawar during 2019.

Wax moth culture

Greater Wax moth (Galleria mellonella) cultures were developed using old, infested frames recovered from bee colonies at ARI Tarnab, Peshawar. These frames provided a consistent and reliable source of wax moth culture for the experiments.

Rearing and testing of development on different cultures

The trials for rearing and treatments were started in first week of April 2019 and ended in June 2019. First instar larvae were selected from the culture. Ten larvae were assigned to each treatment and placed in separate containers enclosed with mesh. The containers were made of plastic and were then placed in an incubator (Infitek-ICB Series, China) set to three temperatures: one group at 25 °C, second at 30 °C, and third at 35 °C to observe the effects of temperature variation. At each temperature, four types of diets were provided as food sources: new wax (new prepare wax sheets), old wax (one year old frame wax), a mixture of old and new wax (50:50 of both), and paper wasp nests (abandoned nests rom trees and old buildings).

Treatments for wax moth control

Different larval instars were separated for experimental purposes, specifically the 1st, 3rd, and 5th instars. Ten larvae from each instar stage were selected for each treatment group. They were placed in individual plastic containers (7x4.75x2.5 inches), which were enclosed with mesh. These containers were then kept in an incubator, where various treatments were applied. Temperature (25±2°C) and humidity at (60+5%) was maintained according to the recommended laboratory standards for wax moth rearing (Ellis et al., 2013). These treatments were applied T1= Cold treatment for three hours, T2= 5% neem spray (extracted from need seeds) on larva, T3= Marigold extraction (flowers extraction) spray and T4= control no treatment was applied. The mortality of wax moth larvae was recorded after 24 hours, 48 hours, 72 hours and 7days.

Statistical analysis

The data were recorded for rearing and treatments trials and means were calculated, the mean were subject to statistical analysis using computer program Statistix-10.

Results

Development under varying feeds and temperatures

The G. mellonella development on four diets varied at 25 °C as presented in Figure 1. Significant differences were recorded in development on different feeds. The development was completed in significantly less time on old wax (54 days), combination of old and new wax (58 days) as compared to new wax (62 days) however development on paper nest was was slow (73 days).

 

 

The development of the wax moth on four diets varied at 30 °C as presented in Figure 2. Significant different were recorded in development on different feeds. The development was completed in significantly less time on old wax (40 days), combination of old and new wax (42 days) as compared to new wax (50 days) however development on paper nest was slow (60 days).

The development at 35° showed accelerated development as compared with 25 °C and 30°C. The development of the wax moth on four diets varied at 35 °C as presented in Figure 3. Significant differences were recorded in development on different feeds. The development was completed in significantly less time on old wax (28 days), combination of old and new wax (33 days) as compared to new wax (37 days) however development on paper nest was slow (40 days).

 

Management of wax using different control strategies

1st instar larvae: Figure 4 illustrates the percentage of dead wax moth larvae (1st instar) under various treatments. After 24 hours, the highest mortality was observed in the cold treatment (T1), with 10 dead larvae, while no mortality was recorded in the control group (T4). At 48 hours, complete mortality (all larvae dead) was again recorded in the cold treatment (T1). In contrast, the lowest mortality was observed in both the marigold treatment (T3) and the control (T4), with only one dead larva in each. After 72 hours, cold treatment (T1) continued to show the highest mortality (5 larvae), whereas the control (T4) had the lowest. By the seventh day, T1 (cold treatment) still exhibited the highest mortality (10 dead larvae), while the control group (T4) had the lowest, with only 1 larva dead.

 

3rd instar larvae: The percentage of dead wax moth larvae (3rd instar) due to different treatments is presented in Figure 5. After 24 hours, the highest mortality (2 larvae) was observed in both the cold treatment (T1) and the marigold treatment (T3), while no mortality was recorded in the control group (T4). After 48 hours, the same trend continued, with the highest mortality (3 larvae) again observed in cold (T1) and marigold (T3), and no mortality in the control. After 72 hours, the maximum deaths (3 larvae) reported in the neem (T2) and marigold (T3) treatments. After 7 days, the maximum mortality (5 larvae each) was recorded in cold (T1), neem (T2), and marigold (T3) treatments. The minimum mortality (1 larvae) in the control group (T4).

 

 

5th instar larvae: Figure 6 shows how many wax moth larvae (5th instar) died under different treatments at different times. After 24 and 48 hours, only the neem treatment (T2) caused any deaths, with 2 larvae dying. No larvae died in the other treatments (cold (T1), marigold (T3), and control (T4). After 72 hours, the control group (T4) had only one dead larvae. After 7 days, the highest number of deaths was again in the neem treatment (T2), with 7 larvae dead. The cold (T1) and marigold (T3) treatments had the lowest, with 2 larvae dead in each. Only one dead larvae was recorded in control (T4).

Discussion

The current study reported that temperature has great impact on the life cycle of G. mellonella. At 25°C the development took longer time as compared to 30°C and 35°C. Similar results were also has been reported by Hanumanthaswamy et al. (2013), who reported that the optimum temperature for wax moth rearing under lab condition ranges from 28°C and 34°C. Kwadha et al. (2017) also reported that temperature below than 25°C and above 35°C can affected life cycle of this pest. At this range of temperature, the metabolism remains at peak and is critical for development, growth rate and reproduction. So, the temperature has an important role in larval development, pupation and adult emergence. The humidity during the experiments were kept constant at 60± 5 %. The humidity is also an important factor in rearing of the wax moth under laboratory conditions. Earlier studies of Ranjbar-Aghdam et al. (2015) and Borah et al. (2024) supported our results that humidity ranges from 50±10% and temperature ranges from 28-35°C are ideal for mass rearing of wax moth under laboratory conditions. Krams et al. (2015) reported that diets with varying compositions affect the life cycle of the moth due to differences in their nutritional quality. Some diets promote faster and earlier development compared to others. Temperature also plays a crucial role in the development of the wax moth across different diets.

In the trials, cold treatment, neem extract spray, and marigold extract were applied at different larval stages of the wax moth to assess their effectiveness in causing mortality. Previous findings by Gulati and Kaushik (2004) indicated that early instar larvae of the wax moth are susceptible to cold treatment; however, this method is less effective against later larval stages, which show higher survival rates under cold conditions. Cold treatment remains a simple and effective method for managing early larval stages of moth in wax storage. In the current study, neem extract also demonstrated high effectiveness, resulting in significant mortality across all larval stages. These findings are consistent with the results of Elbehery et al. (2016), who reported neem’s efficacy in controlling wax moth larvae. Neem extract showed promising results both in laboratory and field conditions, with significantly higher larval mortality.

Conclusions and Recommendations

The temperature and diets influenced the life Cyle of G. mellonella. Optimal growth was observed on old wax at temperatures of 30°C and 35°C. Early instar larvae in infested frames can be effectively managed using cold treatment for 3–5 hours. However, later instar stages are more resistant to cold, requiring the implementation of proper management techniques for effective control. The beekeeper should used pollen diets in dearth period to improve the health and brood rearing of colonies.

Acknowledgment

The authors thanked the Entomological research program at ARI Tarnab for their support.

Novelty Statement

The paper provides insightful information on rearing wax moths on different diets and includes additional methods for their management.

Author’s Contribution

Hussain Ali: Conducted the experiments, collected the data and submitted the paper for publication.

Mirzakhmedov Muhammadjon Fazliddin Ogli: Did the analysis.

Saparov Kuanishbay Abdirakhmanovich, Sultanov Esbosin Kosimbetovich and Embergenov Muratbay Amanbaevich: Wrote this first draft.

Generative AI and AI-assisted technology statement

In this paper, AI was used primarily for correcting grammar and improving sentence structure only.

Conflict of interest

The authors have declared no conflict of interests.

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