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Mating behaviour of the small hive beetle, 
Aethina tumida. 
ZEN 363 Practical 2 
Pitswane M.L 
10294547 
Department of Zoology & Entomology, University of Pretoria, Pretoria, 0002
Abstract 
In many insect species females commonly mate with more than one male. In such events, one 
must consider if there is a selection advantage to first male mating, second male mating, or to 
either. The objective of the current study was to analyse the mating behaviour in Aethina 
tumida, small hive beetles under laboratory conditions. We found that males and females of 
the small hive beetles did not differ in their body size (head, length, and width). It was also 
found that there was a negative correlation between size and mounting-copulation (P<0.05); 
however a positive correlation was observed between size and mounting duration (P>0.05). 
No relationship was observed between mounting and copulation frequency with antennating 
time (R=0.02). It was found that experiment one and two differed significantly in terms of 
mounting duration with relation to length, width and size. In addition, there was a significant 
difference in terms of mounting frequency for length and width. There is too much 
consistency in the data collected from this experiment, and we therefore suggest that more 
experiments should be conducted and we advise future researchers to pay attention to detail. 
Introduction 
Sexual selection is defined as natural selection arising through preference by one sex 
for certain characteristics in individuals of the other sex. Mate choice, a component of sexual 
selection, is an evolutionary process in which selection of a mate depends on the 
attractiveness of his or her traits. Sexual selection may be influenced by length, and body size 
may have a large influence on male-male competition (Forslund 2000). It can be found that 
male-male competition may determine which males are able to access females, and further, 
female choice among available males (Forslund 2000). 
Previous studies have found that in many species females commonly mate with more 
than one male. In such events, one must consider if there is a selection advantage to first male 
mating, second male mating, or to neither (Ginsberg and Huck 1989). If a single female had 
to mate with sat male one (P1) and later male two (P2), the probability of success of sperm 
from P2 can be low (first male precedence as seen in spiders, Austad 1984), or it can be high 
(as seen in many insects, Parker and Simmons 1991). 
In this experiment we look at the mating behaviour of small hive beetles, Aethina 
tumida Murray (Coleoptera: Nitidulidae). These species are native to sub-Saharan Africa, and 
are considered serious pests of honeybees in countries such as Canada and Australia (Hood 
2004). Honeybees are economically valuable to man- in US alone the European honeybee, 
Apis mellifera Lepeletier (Hymenoptera: Apidae) pollinates about US$215 billion worth of
crops (Gallai et.al 2009). The knowledge obtained from this paper can be used in Integrated 
Pest Management to work against small hive beetles and ensure less damage to honeybees. 
The current study looked at the beetle behaviour of males under no competition vs. 
competition. The objectives of the study was to check if there is a relationship between size 
and copulation and mounting time, (2) to test if there is correlation between antennating and 
copulation/mounting frequency or time, and (3) compare the general behaviour performance 
of each male when they are alone with the female (experiment one) vs. when the other male 
is present (experiment two). Our null hypothesis is that there will be significant difference in 
male behaviour between experiment one and experiment two. 
Methods and Materials 
Experiment 
Male and female beetles were collected from the Department of Zoology & Entomology, 
University of Pretoria. The insects were maintained in the fridge prior to experiment. The 
females were marked using paint-like marker on their head. One male (P1) was placed inside 
a petri-dish with the female and their behavioural pattern was observed for 10 minutes. After 
10 minutes the first male was replaced with the second male (P2), and the same observations 
were made. In the second part of the experiment, all beetles, male one, male two and the 
female were all in one petri-dish together for 10 minutes, and their behavioural pattern was 
observed. 
In each case, the following behaviours were noted: mounting, antennating, standing, walking, 
and walk-over, grooming, and copulation. We assumed that mounting is a proxy for mating 
in addition to copulation. 
Data analysis 
All analysis was conducted in Microsoft Excel (Microsoft 2010) and IBM SPSS (Statistics 
22). A correlation under case-wise deletion was used to analyse relationship between size and 
copulation and mounting time as well as frequency. To determine if there was correlation 
between antennating, mounting and copulation, a correlation case-wise deletion analysis was 
conducted and a simple scatter plot graph was done. To compare the behavioural patterns 
(copulation time, mounting duration, and mounting frequency) of each male between the two 
experiments in relation to their size (length, width and head), a Wilcoxon matched pairs test
was conducted. The Wilcoxon test is a non-parametric hypothesis test used for comparing 
related samples, and it serves as an alternative for data that is not normally distributed. 
In total 49 beetles, 22 of which were males and 18 of which were females. 
Results 
Body size in males and females 
It was found that on average the body length, body width and head size of male and female 
subjects did not differ (Table 1). 
Table 1: The average sizes of the length, width and head of the female and male Aethina tumida, small hive 
beetles. 
Beetles Length Width Head 
Females 6.3 3.8 1.3 
Males 6.7 3.8 1.7 
In the first correlation test analysing the relation between size and mounting-copulation, we 
found that there was a significant correlation for size and mounting duration (P<0.05), 
however the opposite was observed for mounting frequency (P>0.05) (Figure 1 and Figure 2). 
Mount duration (sec) 
Small Size category Big 
Figure 1: Correlation between mounting duration and size category of male Aethina tumida species. 
The males were classified as either small or big based on their size, and therefore the data was not 
continuous. Data was obtained from both experiments where (1) each male was given ten minutes 
with the female alone, and (2) both males and the females were together in one petri-dish. N=16
Small Size category Big 
Mount frequency 
Figure 2: Correlation between mounting frequency and size category of male Aethina tumida species. The 
males were classified as either small or big based on their size, and therefore the data was not continuous. 
Data was obtained from both experiments where (1) each male was given ten minutes with the female 
alone, and (2) both males and the females were together in one petri-dish. N=16 
Correlation between antennating and mounting/copulation frequency and duration 
Analysis of mounting and copulation frequency combined against antennating time revealed 
that there was no relationship between the two parameters (R族=0.02) (Fig.1). 
y = -0.1538x + 95.747 
R族 = 0.0131 
600 
500 
400 
300 
200 
100 
0 
0 100 200 300 400 500 
Mounting/Copulation time (sec) 
Antennating time (sec) 
Figure 3: A scatter plot showing the relationship between antennating time and mounting/copulation time 
of Aethina tumida male species. Data was collected from 22 males that were singularly exposed to one 
female (different for each male) for ten minutes each.
Comparison of male behaviour between experiment one (single male and female) and 
experiment two (two males and one female) 
It was found that experiment one and two differed significantly in terms of mounting duration 
with relation to length, width and size (Table 2). In addition, there was a significant 
difference in terms of mounting frequency for length and width. Data for experiment two for 
copulation time was missing. 
Table 2: Comparison of mating behaviour patterns of Aethina tumida males when 
exposed to one female (exp 1) and when exposed to one female in presence of another 
male (exp 2). Those behaviours that showed a significant difference between the two 
experiments are highlighted (dark writing). (-) indicates no data. 
Mount duration Mount frequency Copulation time 
Exp 1 Exp 2 Exp 1 Exp 2 Exp 1 Exp 2 
Length 0.313464 0.025094 0.000276 0.000112 0.575487 - 
Width 0.287338 0.021679 0.006947 0.001507 0.575487 - 
Head 0.145401 0.005685 0.663197 0.687409 0.575487 - 
Discussion 
P-values 
In this species, we found that males and females do not differ very much in terms of their 
body size (head, length, and width). Given these results, we cannot expect size to have a 
significant difference on mating behaviours; however care must be taken that this lack of 
difference is observed between the sexes, there might be a large variation within the same 
sex. Mounting duration showed a negative correlation with size, and surprisingly, mounting 
frequency showed a positive correlation. In this study there were a lot of inconsistencies with 
regards to the data and this proved a difficulty in data analysis. For example, measurements 
were not consistent, in experiment two we failed to record copulation duration. 
In comparing the two experiments to find out if presence of second male played a role 
in the mating behaviour, we found that there was no difference. Whether the male was alone 
with the female or with yet other male, mating behaviours were similar, in other words, we 
could say that there was no competition. However, in some individuals we observed some 
dominance during the second experiment, of which varied from the larger male to the smaller 
male within the enclosure.
In certain individuals we observed that other males would mount on the other male 
instead of the female. Switzer et.al (2014) suggests that the reasons behind this observation 
may lie in understanding the selection pressures resulting from the mating systems of the 
species. In addition, we did not observe any aggressive competition between the beetles. 
However, the males did sometimes attempt to take over ecen after the first male had 
mounted/copulated with the female. Kruse and Switzer (2007) contest that even though such 
events may last longer, the chances of successfulness is minimal. Thus, a males best option 
may be to be the first male and find an available female (Switzer et.al 2014)
References 
Bonduriansky, R. 2001. The evolution of male mate choice in insects: a synthesis of ideas 
and evidence. Biology Review 76: 305-339 
Fisher, R. A. 1930. The Genetical Theory of Natural Selection. Oxford, Clarendon Press 
Forslund P. 2000. Male-male competition and large size mating advantage in European 
earwigs, Forficula auricularia. Animal Behaviour 59:753-762. 
Gallai N., Salles J.M, Settele J., Vaissiere B.E. 2009. Economic valuation of the vulnerability 
of world agriculture confronted with pollinator decline. Ecological Economics 68(3):810821 
Ginsberg, J.R., Huck, U.W. 1989.Sperm competition in mammals. Trends Ecological 
Evolution 4: 74-79 
Kruse, K.C., Switzer, P.V. 2007. Physical contests for females in the Japanese beetle, 
Popillia japonica. Journal of Insect Sciences 7:34 
Switzer, P.V., Forsythe, P.S., Krusel, K.C. 2014. Male-male mounting and the unreliability of 
body size as a character for mate choice in male Japanese beetles (Popillia japonica 
Newman). Zoological Studies 53:53 
Appendix 
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Mating behaviour of the small hive beetle, Aethina tumida.

  • 1. Mating behaviour of the small hive beetle, Aethina tumida. ZEN 363 Practical 2 Pitswane M.L 10294547 Department of Zoology & Entomology, University of Pretoria, Pretoria, 0002
  • 2. Abstract In many insect species females commonly mate with more than one male. In such events, one must consider if there is a selection advantage to first male mating, second male mating, or to either. The objective of the current study was to analyse the mating behaviour in Aethina tumida, small hive beetles under laboratory conditions. We found that males and females of the small hive beetles did not differ in their body size (head, length, and width). It was also found that there was a negative correlation between size and mounting-copulation (P<0.05); however a positive correlation was observed between size and mounting duration (P>0.05). No relationship was observed between mounting and copulation frequency with antennating time (R=0.02). It was found that experiment one and two differed significantly in terms of mounting duration with relation to length, width and size. In addition, there was a significant difference in terms of mounting frequency for length and width. There is too much consistency in the data collected from this experiment, and we therefore suggest that more experiments should be conducted and we advise future researchers to pay attention to detail. Introduction Sexual selection is defined as natural selection arising through preference by one sex for certain characteristics in individuals of the other sex. Mate choice, a component of sexual selection, is an evolutionary process in which selection of a mate depends on the attractiveness of his or her traits. Sexual selection may be influenced by length, and body size may have a large influence on male-male competition (Forslund 2000). It can be found that male-male competition may determine which males are able to access females, and further, female choice among available males (Forslund 2000). Previous studies have found that in many species females commonly mate with more than one male. In such events, one must consider if there is a selection advantage to first male mating, second male mating, or to neither (Ginsberg and Huck 1989). If a single female had to mate with sat male one (P1) and later male two (P2), the probability of success of sperm from P2 can be low (first male precedence as seen in spiders, Austad 1984), or it can be high (as seen in many insects, Parker and Simmons 1991). In this experiment we look at the mating behaviour of small hive beetles, Aethina tumida Murray (Coleoptera: Nitidulidae). These species are native to sub-Saharan Africa, and are considered serious pests of honeybees in countries such as Canada and Australia (Hood 2004). Honeybees are economically valuable to man- in US alone the European honeybee, Apis mellifera Lepeletier (Hymenoptera: Apidae) pollinates about US$215 billion worth of
  • 3. crops (Gallai et.al 2009). The knowledge obtained from this paper can be used in Integrated Pest Management to work against small hive beetles and ensure less damage to honeybees. The current study looked at the beetle behaviour of males under no competition vs. competition. The objectives of the study was to check if there is a relationship between size and copulation and mounting time, (2) to test if there is correlation between antennating and copulation/mounting frequency or time, and (3) compare the general behaviour performance of each male when they are alone with the female (experiment one) vs. when the other male is present (experiment two). Our null hypothesis is that there will be significant difference in male behaviour between experiment one and experiment two. Methods and Materials Experiment Male and female beetles were collected from the Department of Zoology & Entomology, University of Pretoria. The insects were maintained in the fridge prior to experiment. The females were marked using paint-like marker on their head. One male (P1) was placed inside a petri-dish with the female and their behavioural pattern was observed for 10 minutes. After 10 minutes the first male was replaced with the second male (P2), and the same observations were made. In the second part of the experiment, all beetles, male one, male two and the female were all in one petri-dish together for 10 minutes, and their behavioural pattern was observed. In each case, the following behaviours were noted: mounting, antennating, standing, walking, and walk-over, grooming, and copulation. We assumed that mounting is a proxy for mating in addition to copulation. Data analysis All analysis was conducted in Microsoft Excel (Microsoft 2010) and IBM SPSS (Statistics 22). A correlation under case-wise deletion was used to analyse relationship between size and copulation and mounting time as well as frequency. To determine if there was correlation between antennating, mounting and copulation, a correlation case-wise deletion analysis was conducted and a simple scatter plot graph was done. To compare the behavioural patterns (copulation time, mounting duration, and mounting frequency) of each male between the two experiments in relation to their size (length, width and head), a Wilcoxon matched pairs test
  • 4. was conducted. The Wilcoxon test is a non-parametric hypothesis test used for comparing related samples, and it serves as an alternative for data that is not normally distributed. In total 49 beetles, 22 of which were males and 18 of which were females. Results Body size in males and females It was found that on average the body length, body width and head size of male and female subjects did not differ (Table 1). Table 1: The average sizes of the length, width and head of the female and male Aethina tumida, small hive beetles. Beetles Length Width Head Females 6.3 3.8 1.3 Males 6.7 3.8 1.7 In the first correlation test analysing the relation between size and mounting-copulation, we found that there was a significant correlation for size and mounting duration (P<0.05), however the opposite was observed for mounting frequency (P>0.05) (Figure 1 and Figure 2). Mount duration (sec) Small Size category Big Figure 1: Correlation between mounting duration and size category of male Aethina tumida species. The males were classified as either small or big based on their size, and therefore the data was not continuous. Data was obtained from both experiments where (1) each male was given ten minutes with the female alone, and (2) both males and the females were together in one petri-dish. N=16
  • 5. Small Size category Big Mount frequency Figure 2: Correlation between mounting frequency and size category of male Aethina tumida species. The males were classified as either small or big based on their size, and therefore the data was not continuous. Data was obtained from both experiments where (1) each male was given ten minutes with the female alone, and (2) both males and the females were together in one petri-dish. N=16 Correlation between antennating and mounting/copulation frequency and duration Analysis of mounting and copulation frequency combined against antennating time revealed that there was no relationship between the two parameters (R族=0.02) (Fig.1). y = -0.1538x + 95.747 R族 = 0.0131 600 500 400 300 200 100 0 0 100 200 300 400 500 Mounting/Copulation time (sec) Antennating time (sec) Figure 3: A scatter plot showing the relationship between antennating time and mounting/copulation time of Aethina tumida male species. Data was collected from 22 males that were singularly exposed to one female (different for each male) for ten minutes each.
  • 6. Comparison of male behaviour between experiment one (single male and female) and experiment two (two males and one female) It was found that experiment one and two differed significantly in terms of mounting duration with relation to length, width and size (Table 2). In addition, there was a significant difference in terms of mounting frequency for length and width. Data for experiment two for copulation time was missing. Table 2: Comparison of mating behaviour patterns of Aethina tumida males when exposed to one female (exp 1) and when exposed to one female in presence of another male (exp 2). Those behaviours that showed a significant difference between the two experiments are highlighted (dark writing). (-) indicates no data. Mount duration Mount frequency Copulation time Exp 1 Exp 2 Exp 1 Exp 2 Exp 1 Exp 2 Length 0.313464 0.025094 0.000276 0.000112 0.575487 - Width 0.287338 0.021679 0.006947 0.001507 0.575487 - Head 0.145401 0.005685 0.663197 0.687409 0.575487 - Discussion P-values In this species, we found that males and females do not differ very much in terms of their body size (head, length, and width). Given these results, we cannot expect size to have a significant difference on mating behaviours; however care must be taken that this lack of difference is observed between the sexes, there might be a large variation within the same sex. Mounting duration showed a negative correlation with size, and surprisingly, mounting frequency showed a positive correlation. In this study there were a lot of inconsistencies with regards to the data and this proved a difficulty in data analysis. For example, measurements were not consistent, in experiment two we failed to record copulation duration. In comparing the two experiments to find out if presence of second male played a role in the mating behaviour, we found that there was no difference. Whether the male was alone with the female or with yet other male, mating behaviours were similar, in other words, we could say that there was no competition. However, in some individuals we observed some dominance during the second experiment, of which varied from the larger male to the smaller male within the enclosure.
  • 7. In certain individuals we observed that other males would mount on the other male instead of the female. Switzer et.al (2014) suggests that the reasons behind this observation may lie in understanding the selection pressures resulting from the mating systems of the species. In addition, we did not observe any aggressive competition between the beetles. However, the males did sometimes attempt to take over ecen after the first male had mounted/copulated with the female. Kruse and Switzer (2007) contest that even though such events may last longer, the chances of successfulness is minimal. Thus, a males best option may be to be the first male and find an available female (Switzer et.al 2014)
  • 8. References Bonduriansky, R. 2001. The evolution of male mate choice in insects: a synthesis of ideas and evidence. Biology Review 76: 305-339 Fisher, R. A. 1930. The Genetical Theory of Natural Selection. Oxford, Clarendon Press Forslund P. 2000. Male-male competition and large size mating advantage in European earwigs, Forficula auricularia. Animal Behaviour 59:753-762. Gallai N., Salles J.M, Settele J., Vaissiere B.E. 2009. Economic valuation of the vulnerability of world agriculture confronted with pollinator decline. Ecological Economics 68(3):810821 Ginsberg, J.R., Huck, U.W. 1989.Sperm competition in mammals. Trends Ecological Evolution 4: 74-79 Kruse, K.C., Switzer, P.V. 2007. Physical contests for females in the Japanese beetle, Popillia japonica. Journal of Insect Sciences 7:34 Switzer, P.V., Forsythe, P.S., Krusel, K.C. 2014. Male-male mounting and the unreliability of body size as a character for mate choice in male Japanese beetles (Popillia japonica Newman). Zoological Studies 53:53 Appendix Available on :