# Impact of different constant temperatures on the survival of honey bee, Apis mellifera (Hymenoptera: Apidae)

Muhammad Sarmad, Pavel Ryšánek

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Muhammad Sarmad, Pavel Ryšánek. Impact of different constant temperatures on the survival of honey bee, Apis mellifera (Hymenoptera: Apidae). Apidologie, 2025, 56 (4), pp.68. 10.1007/s13592-025-01191-y. hal-05160427

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# Impact of different constant temperatures on the survival of honey bee, *Apis mellifera* (Hymenoptera: Apidae)

Muhammad SARMAD¹, and Pavel RYŠÁNEK¹

¹ Department of Plant Protection, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences, Prague 16500, Czech Republic

Received 13 November 2024 – Revised 19 May 2025 – Accepted 4 June 2025

## Abstract

Honey bees play a crucial role as beneficial insects for producing honey and pollinating crops. *Apis mellifera* (Linnaeus) (Hymenoptera: Apidae) is one of the most important species of honey bees globally. Insects experience various temperature stresses in their natural habitat that differ from their typical temperature range. Being an important pollinator, the current study is designed to investigate the effect of temperature stress during summer and winter on bees' survival, recovery rate, and longevity. To check their sensitivity, the worker bees were exposed to summer (31 to 19 °C) and winter (12 to 0 °C) temperatures. Different temperatures in summer were significantly influenced the survival rate and the time required to achieve 50% mortality. The 50% mortality was observed in 13.79 days at 31 °C and gradually decreased with the decrease in temperature. The recovery rate and longevity of the bees decreased, when exposed to cold temperatures, 12 to 0 °C for 1, 2, and 3 h. Both aspects eventually decreased with the increase in exposure duration. Based on the results, it is concluded that the survival rate, recovery rate, and longevity of the bees were greatly affected by the change in temperature.

## Honey bees / Summer / Winter / Survival rate / Recovery rate / Longevity

### 1. INTRODUCTION

Honey bees, *Apis mellifera* L. (Hymenoptera: Apidae), are an important pollinator for agricultural ecosystems (Calderone 2012; Hackett 2004; Kevan et al. 2007; Southwick and Southwick Jr 1992). Honey bees remain vital to the global food supply, even with the presence of competing pollinators (McGregor 1976; Watanabe 1994). The decline of these pollinators can result in a decrease of up to 90% in crop yields for certain fruits, seeds, and nuts (Southwick and Southwick Jr 1992). In the absence of wild bees, farmers often rely on honey bee hives to pollinate their crops.

Honey bees are versatile, cost-effective, and convenient when compared to the management of various wild bee species; however, for certain crops, they are not the most effective pollinators for each individual flower (Parker et al. 1987; Richards 1996; Torchio 1990). The number of well-managed honey bee colonies is decreasing in certain parts of the globe (Delaplane and Mayer 2000; Klein et al. 2007; Matheson 1996; Williams et al. 1991) due to parasitic mites, *Varroa destructor* (Mesostigmata: Varroidae) (Downey and Winston 2001), the application of different pesticides, a reduction in younger beekeepers worldwide, and the effects of climate change (Hristov et al. 2021; Ingram et al. 1996).

Fluctuations in environmental factors are recognized as a stressor for insect populations worldwide (Hoffmann and Sgrò 2011). The

<figure><img src="image_2.png" /></figure>

<footnote>Corresponding author: M. Sarmad, sarmad@af.czu.cz
Communicated by James Nieh.</footnote>
temperature range within *A. mellifera* bee hives is between 33.5 and 35 °C. Temperature variations at the hive entrance occur due to change in environmental conditions (Fahrenholz et al. 1989). Many studies have shown that honey bee capped brood raised at suboptimal temperatures can have negative effect on adult bees such as deformed wings, legs, and abdomen (Himmer 1927, 1932). When pupae of *A. mellifera scutellata* are exposed at 20 °C for 96 h, they show a high percentage of deformed wings (Chacon-Almeida et al. 1999). Adult bees raised at lower temperatures as brood also show differences in behavior, learning, and memory (Jones et al. 2005).

There is a shortage of comprehensive studies examining how varying summer and winter temperatures affect the sensitivity of *A. mellifera* in the Czech Republic. In Prague, the temperature has been varying, with the maximum and minimum temperatures recorded each day throughout the summer months (May to October 2024), which might affect the survival of pollinators. Considering the importance of *A. mellifera* in the Czech Republic, this study sought to determine the survival rate, recovery rate, and longevity at different constant temperatures.

# 2. MATERIAL AND METHODS

## 2.1. Honey bees

Honey bee hives were managed at the Green House Site, Department of Plant Protection, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences, Prague. This area is full of flowering plants to get pollen and nectar and is free from pesticides. All managing practices were done according to the standard beekeeping rules (Flottum 2024). Bee hives were evaluated after an interval of 5 to 7 days to monitor the normal activity of the bees, eliminate new queen cells to prevent swarming, and guard against diseases and parasites.

A brood frame containing at least five emerging worker bees was placed into the box with wire mesh walls and moved into the incubator (33 ± 1 °C; 65 ± 3 RH; Photoperiod L16: D8 h) to stimulate natural environmental conditions.

## 2.2. Effect of summer temperatures on survival rate of worker bees

Newly emerging five hundred worker bees (<1 day old) were shifted into twenty plastic boxes (1 box = 16×11×5.5 cm) with muslin cloth on the top for aeration. Each treatment was replicated five times, with 20 bees in each replication. The pollen was placed with water and a 65% sugar solution in each box for the first seven days. After 7 days, two pady pods with plastic tubes (15 ml) were used in each box to provide a 65% sugar solution and water as a diet. The bees in boxes were placed at 19 °C, 23 °C, 27 °C, and 31 °C in the growth chamber to check the survival rate and lethal time 50% (Ltime50). The control bees were placed at 33 ± 1 °C to check the normal movement of the bees. The bees were kept in each mentioned temperature until 100% mortality. The mortality was observed after 24 h.

## 2.3. Effect of winter temperatures shock on recovery rate and longevity of worker bees

Newly emerging fifteen hundred bees (hundred bees in each treatment) were kept at 33 ± 1 °C for 7 days before cold shock. The pollen was placed in each box with water and 65% sugar solution for the first 7 days. The bees (7 days old) in plastic boxes were placed at 12 °C, 8 °C, 4 °C, and 0 °C in the growth chamber for 1, 2, and 3 h (Sarmad et al. 2020a). The control bees were placed at 33 ± 1 °C for comparison in each exposure duration. Each treatment was replicated five times, with 20 bees in each replication. After exposure to the particular
<table><caption>**Table I** Lethal time (Ltime50) and confidence interval (95% CI) for worker bees at four summer temperatures of Prague</caption><thead><tr><th>Temp (°C)</th><th>d.f.</th><th>Slope (±SE)<sup>a</sup></th><th>Ltime50 (95% CI)</th><th>χ<sup>2</sup></th><th>P</th></tr></thead><tbody><tr><td>31</td><td>3</td><td>4.00 ± 0.36</td><td>13.79 (11.79–15.78)</td><td>42.33</td><td>&lt;0.001</td></tr><tr><td>27</td><td>3</td><td>3.53 ± 0.33</td><td>10.77 (9.15–12.31)</td><td>32.54</td><td>&lt;0.001</td></tr><tr><td>23</td><td>3</td><td>3.16 ± 0.30</td><td>9.12 (7.51–10.67)</td><td>36.84</td><td>&lt;0.001</td></tr><tr><td>19</td><td>3</td><td>3.09 ± 0.27</td><td>6.43 (5.55–7.32)</td><td>21.2</td><td>&lt;0.001</td></tr></tbody></table>

temperature for the mentioned hours, the bees were moved to an incubator at 33 ± 1 °C for 2 h (Sarmad et al. 2020b) to check the recovery rate. The same cold shock stressed bees were later kept at 33 ± 1 °C until 100% mortality to check their longevity.

## 2.4. Statistical analysis

The percentage mortality was calculated by using probit analysis in POLO-Plus software (LeOra 2003). Time mortality data (in days) were analyzed to estimate the 50% lethal time (Ltime50) with 95% confidence intervals.

The mean comparison “ANOVA” for recovery rate and longevity was performed, followed by the LSD test by randomized complete design (CRD), using the Statistical Analysis Software (SAS Institute 2002). The SPSS software package (version 21 for Windows, SPSS Inc., Chicago, IL, USA) was used to make Kaplan-Meier survival curve.

# 3. RESULTS

## 3.1. Effect of summer temperatures on survival rate of worker bees

The lethal time (Ltime50) was decreased 13.79, 10.77, 9.12, and 6.43 days when exposed to 31, 27, 23, and 19 °C, respectively (Table I). The data was statistically different at all temperatures, i.e.,

19 °C (df=3, *F*=115.88, *P*<0.001), 23 °C (df=3, *F*=59.01, *P*<0.001), 27 °C (df=3, *F*=116.29, *P*<0.001), and 31 °C (df=3, *F*=89.26, *P*<0.001). The Kaplan-Meier survival analysis presented in the plot illustrates the cumulative survival probabilities of four different temperature treatments: 19, 23, 27, and 31 °C, over different time periods. The survival rate of bees decreased with the decline in temperature (Figure 1). Among all treatments, individuals maintained at 31 °C exhibited the highest survival rate throughout the experimental duration. The individuals exposed to 19 °C demonstrated the lowest survival rate. The 23 °C and 27 °C treatments showed intermediate survival outcomes, with 27 °C supporting relatively better survival than 23 °C (Figure 2). The coefficient of determination (r²) range was recorded as high, from 0.99 to 0.96 (Figure 3).

## 3.2. Effect of winter temperatures shock on recovery rate and longevity of worker bees

The percentage of recovery rate declined with the increase in exposure duration (1, 2, and 3 h) to low temperature stress. The maximum recovery rate was 81, 78, and 66% at 12 °C, while the minimum recovery rate was 35, 17, and 8% observed at 0 °C after 1, 2, and 3 h of exposure, respectively. The data was statistically (df=14, *F*=89.05, *P*<0.001) different after exposure to different durations. The maximum number of bees was recovered after 1 h of

<figure><img src="image_3.png" /></figure>

<footnote><sup>a</sup>Estimated parameter of Probit analysis</footnote>
<figure><img src="image_4.png" /><figcaption>**Fig. 1** Temperature recorded on a daily basis at Green House Site, Czech University of Life Sciences, Prague, from May to October 2024. A May, B June, C July, D August, E September, F October</figcaption></figure>

exposure as compared to 2 and 3 h. The 100% of bees were alive in the control group at 33 °C (Figure 4).

The longevity of bees was decreased due to short-term cold shock. The maximum longevity was observed in the control at 33 °C. The higher longevity was observed at 18.6, 15, 10.6, and 6.6 days after 1 h (df=4, F=32.20,

P<0.001) cold shock exposure as compared to 2 h (15.2, 13, 8.4, and 2.8 days) (df=4, F= 36.75, P<0.001) and 3 h (8.6, 3.8, 1.6 and 0.4 days) (df=4, F= 114.37, P<0.001) at 12, 8, 4, and 0 °C, respectively. Meanwhile, the minimum longevity was observed at 0 °C (Figure 5).

<figure><img src="image_5.png" /></figure>
<figure><img src="image_6.png" /><figcaption>**Fig. 2** Survival rate of the worker bees from 0 to 27 days after exposure to summer temperatures of Prague. Each treatment was replicated five times, with 20 bees in each replication</figcaption></figure>
<figure><img src="image_7.png" /><figcaption>Duration of exposure (days)</figcaption></figure>

**Fig. 3** Line graph representing the linear relationship between the duration of exposure in each temperature and the mortality rate in probit units. **A** 19 °C, **B** 23 °C, **C** 27 °C, and **D** 31 °C
<figure><img src="image_8.png" /><figcaption>**Fig. 4** Recovery rate of worker bees after exposure to winter temperature shock for 1, 2, and 3 h. Each treatment was replicated five times, with 20 bees in each replication. The control bees were placed at 33 ± 1 °C for comparison</figcaption></figure>

<figure><img src="image_9.png" /><figcaption>**Fig. 5** Longevity of worker bees after exposure to winter temperature shock for 1, 2, and 3 h. Each treatment was replicated five times, with 20 bees in each replication. The control bees were placed at 33±1 °C for comparison. The error bars represent the significant difference between exposure durations (1, 2, and 3 h)</figcaption></figure>

## 4. DISCUSSION

A change in temperature significantly impacts the honey bee individuals in terms of their survival rate, recovery rate, and lifespan. The current work was performed to evaluate the effect

of summer and winter temperatures on the honey bee individuals. Different temperatures have a direct effect on the insect fitness (Storey and Storey 2000). The behaviors of insects change until they stay under adverse conditions (Košťál et al. 2001). The survival duration varies for each

<figure><img src="image_10.png" /></figure>
species and can be affected by alterations in envi-
ronmental factors (Danks 1978). Most insects are
inactive or die when exposed to different tem-
perature stress (Staszak and Mutchmor 1973).
In this study, the survival rate of *A. mellifera*
decreased with the decrease in temperature from
31 to 19 °C. Our findings are related to the effect
of an increase in temperature on the foraging of
*Acromyrmex lobicornis* (Hymenoptera: Formi-
cidae). The foraging and walking behaviors of
*A. lobicornis* decreased as temperature increased
(Tizón et al. 2014); this is because *A. lobicornis*
may develop at temperatures of 25 °C, whereas
*A. mellifera* requires a higher temperature of 33
°C or more to develop effectively (Fahrenholz
et al. 1989).

The performance of some insects is totally
different under different conditions. The lon-
gevity of *Phlebotomus papatasi* (Diptera: Psy-
chodidae) was increased when exposed to low
temperatures (Guzmán and Tesh 2000). The
longevity of *A. mellifera* was decreased with the
decrease in temperature. The maximum longev-
ity, 18.6 days, was observed at 12 °C after 1 h of
cold shock and decreased to 15.2 and 8.6 days
with the increase in cold shock duration to 2
and 3 h, respectively. Our finding relates to the
thermal tolerance of *Frankliniella occidentalis*
(Thysanoptera: Thripidae). The longevity of *F.
occidentalis* was decreased due to a decline in
temperature. Males were greatly affected by the
change in temperature compared to females (Li
et al. 2011). It is concluded that ion homeosta-
sis may be involved in longevity that is highly
affected by cold temperature stress (Koštál et al.
2006). A study on *Megachile rotundata* (Hyme-
noptera: Megachilidae) showed low temperature
stress on pupae decreased the flight performance
in males and reduced feeding and activity level
(Bennett et al. 2015). Longevity of adult bees
is a key factor affecting colony health. Short-
lived workers can have a significant impact on
colony health, resulting in a shortage of forag-
ers (Thompson et al. 2007). It will cause earlier
foraging in younger bees, and this acceleration
causes a possible feedback loop to result in col-
ony failure (Perry et al. 2015).

In conclusion, this study evaluates the effect of
summer and winter temperature stress on the sensi-
tivity of *A. mellifera* in Czech Republic. Fluctuation
in temperature has a negative effect on survival of
the bees. The results could be helpful for beekeep-
ers in the management of bees at different tempera-
tures and could also be useful for future thermal and
cryobiology studies on pollinators.

ACKNOWLEDGEMENTS

The authors are thankful to Miloslav Zouhar for providing
the material that was used in the experiment.

AUTHOR CONTRIBUTIONS

MS performed the experiment, analysis, and wrote the
first draft of the manuscript. PR supervised the work.

FUNDING

Open access publishing supported by the institutions par-
ticipating in the CzechELib Transformative Agreement.

DATA AVAILABILITY

The datasets generated during and/or analyzed during
the current study are available from the corresponding
authors on reasonable request.

CODE AVAILABILITY

Not applicable

DECLARATIONS

Ethics approval Not applicable

Consent to participate Not applicable

Consent for publication Not applicable

**Competing interests** The authors declare no competing interests.

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