Journal of Apicultural Research

ISSN: 0021-8839 (Print) 2078-6913 (Online) Journal homepage: http://www.tandfonline.com/loi/tjar20

Increased food supply to all larvae after

dequeening honey bee colonies

J Woyke

To cite this article: J Woyke (1999) Increased food supply to all larvae after
dequeening honey bee colonies, Journal of Apicultural Research, 38:3-4, 117-123, DOI:
10.1080/00218839.1999.11101002

To link to this article: https://doi.org/10.1080/00218839.1999.11101002

Published online: 24 Mar 2015.

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Journal of Apiru/wra/ Research 38(3---4): 117-123 (1999) © 19991BRA

Increased food
supply to all
larvae after
dequeening
honey bee
colonies SUMMARY
Five queenright honey bee (Apis mel/ifera)
colonies were divided into equal queen right and
JWCNKE
queenless subunits, two in the autumn and
Bee Division, Agricultural University- SGGW, three in the spring. Brood food was measured
166 Nowoursynowska, 02-787 Warsaw, Poland from a total of 10 850 cells with larvae identi-
(Received 28 November 1997, fied as 1, 2, 3 and 4 days old. Larvae in queen-
accepted subject to revision 19 May 1998, less subunits received more food than similar
accepted for publication 24 March 1999)
aged larvae in queenright colonies. In the
autumn, 10 larvae, 1 and 4 days old, received
Keywords: Apis mellifera, larvae, royal jelly, on average 12.8 vs. 16.6 mg of food (130%) and
brood food, larval food, queenless colonies 145.4 vs. 163.0 mg (112%) in queenright and
queen less subunits, respectively. Larvae of inter-
mediate ages received intermediate amounts of
food. The largest differences in ratios of food
were noted the day after the subunits were
formed. In queenright colonies, larvae of all ages
received 124% of the nourishment received in
queenless colonies. With time, the amount
decreased to 108%, the sixth day after
dequeening. In the spring, 10 larvae, 1 and 4
days old, received an average of 32.1 vs. 43.9 mg
of food (137%) and 126.3 vs. 144.7 mg (115%)
in queenright and queenless subunits, respec-
tively. Larvae of intermediate ages received
intermediate amounts of food. The largest dif-
ferences in ratios of food were noted the day
after the subunits were formed. In queenright
colonies, larvae of all ages received 175% of the
nourishment in queenless colonies. The largest
increase of 201% was recorded in cells with 1-
day-old larvae, the next day after dequeening.
As time passed, the relative food amount in
queenless colonies decreased to 105% the sev-
enth day after dequeening.
118 Woyke

INTRODUCTION
Many queen honey bees (Apis meHifera) are reared each
year. The most popular method is to rear the queens
in queen less colonies. However, brood feeding behav-
iour of bees after dequeening a colony is not well
known. It is generally known that worker bees con-
struct emergency queen cells after a colony is
dequeened. According to Gontarski (1956), workers
at first alter some worker cells, containing young
female larvae, into queen cells. Under this stimulus,
nurse bees start to supply the larvae with larger
amounts of royal jelly. However, it has not been demon-
strated that this sequence is correct. Kuwabara (1947)
reported that the number of nurse bee visits to cells
containing larvae increased after queen removal.
In the present investigation, the amount of food sup-
plied to larvae in normal colonies and those recently
deprived of a queen was compared.
MATERIALS AND METHODS
Five unrelated queenrightA m. me/lifera hybrid colonies
were divided into otherwise equal queenright, and
queen less subunits, two in the autumn and three in the
spring.
The day after division, brood food was aspirated and
measured from a total of 10 850 cells with larvae iden-
tified as 1, 2, 3 and 4 days old. Larval age was deter-
mined according to drawings by Nelson et a/. (1924).
Larval food was collected with the aid of specially con-
structed pipette tubes. Food from 10 larvae of the
same age was sucked into one tube. Empty and filled
tubes were weighed, and the weight of larval food was
the difference between the weight of filled and empty
tubes. This procedure was repeated each day for lar-
vae of all four ages, six times in the autumn and five
times in the spring. The food was collected over six
days in the autumn and seven days in the spring.
Over several years the time period between dequeen-
ing of bee colonies and the construction of emergency
queen cells was recorded.
One-way analysis of variance (AN OVA) was applied for
the amounts of larval food in queenright (QR) and
queenless (QL) colonies. Autumn and spring results
were analysed separately. To analyse amounts of food
supplied to larvae on particular days, a series of
AN OVA was applied separately for each age of larvae
present in both QR and QL colonies (means in tables
followed by lowercase letters). To analyse overall
means of the amounts of food supplied to larvae of a
particular age during all days of collection, all results
were arranged into eight groups: age 1, 2, 3 and 4 days

0
1 2 3 4 5 6 Mean
124 121 120 114 109 108
D 119 113 112 111 110 108 112
D 119 116 112 111 109 113
lm1 131 130 123 121 126
• 127 124 137 130

FIG. 1. Percentage of the amount of larval food observed in brood cells in queenless colonies in
ratio to that found in queenright colonies in autumn. The curve indicates means for larvae of all
ages. The first row beneath the graph indicates days after dequeening. Data in the table indicate
percentage of food provided.
Larval food in gueenless colonies 119

in each QR and QL colonies (means in tables followed
by capital letters). Two-way ANOVA was applied sep-
arately for each age of larvae in QR and QL colonies.
The comparison model was QR/QL x day of food col-
lection. All means were compared using LSD 0.95 test.
Standard error of the means for each age of larvae sep-
arately in QR and QL colonies were calculated.
RESULTS
Larvae up to three days old received white royal jelly
and older ones received transparent mixed food.
Amo...,t of food supplied in autumn to larvae in
queenright and queenless colonies in subse-
quent days after dequeening Table 1 shows that
in autumn, ten 1-day-old larvae, received 12.8 ± 0.18
mg liS. 16.6 ± 0.42 mg offood in queen right and queen-
less subunits, respectively. Similarly, 4-day-old larvae
received 145.4 ± 0.72 mg 1/S. 163.0 ± 1.06 mg offood,
respectively. Larvae of intermediate ages were supplied
with intermediate amounts of food. The overall mean
amounts of food supplied to larvae of the same age
were significantly higher in queenless than in queen-
right colonies for larvae of all four ages.
In subsequent days after the experiment, larvae of each
age in queenright colonies received mostly similar
amounts of food over the days following collection. Lar-
vae of the same age received significantly more food in
queenless than in queenright colonies over consecu-
tive days after dequeening. However, a tendency was
observed to supply larvae in queenless colonies with
decreasing amounts of food as time passed. This was
proven statistically only for some 4-day-old larvae. Sig-
nificant interaction (QR/QL X days of collection) was
found for those 4-day-old larvae (P 0.0064. Conse- =
quently, the increased amount of larval food in queen-
less colonies over that in queenright colonies
decreased as time passed.
The amount of larval food in queenless colonies was
calculated as a percentage relative to that in queen right
units. Figure 1 shows that the relative amount of food
in queenless colonies was the highest in cells with 1-
day-old larvae at 130%. The percentage gradually
decreased for older larvae, reaching the lowest level of
112% for 4-day-old larvae. This contrasts with the total
amount of larval food, where 4-day-old larvae received
the greatest supply (table 1). In successive days, the
largest increase in ratios of food provided was found
to be the day after dequeening. Larvae of all ages in
queenless colonies received 124% of the nourishment
supplied to larvae in queenright colonies. With time,
the amount decreased to 108% on the sixth day after
dequeening. Thus, as time passed, the amount of food
provided to larvae in queenless colonies approached
that found in the queen right units.
Amount of food supplied in spring to larvae in
queenright and queenless colonies in subse-
quent days after dequeening Table 2 shows that
in the spring ten 1-day-old larvae received on average
32.1 ± 0.74 mg liS. 43.9 ± 1.78 mg of food in queen-
right and queenless subunits, respectively. Similarly, lar-
vae 4 days old received 126.3 ± 3.16 mg 1/S. 144.0 ±
4.89 mg of food, respectively. Larvae of intermediate
ages were supplied with intermediate amounts of food.
The overall amount of food fed to larvae of the same
age was significantly higher in queenless than in queen-
right colonies. The total amount of food supplied in the

TABLE 1. Mean amount offood {mg) in 10 cells with larvae of different age on subsequent
days after dequeening, in two queenright {QR) and two queenless {QL) colonies in autumn
{25-30 August). Each number for a particular day represents a mean of 12 replicates of food
collected from 120 larvae.
Age of QR Days after de9ueening Overall
larvae {days) orQL 1 2 3 4 5 6 mean± s.e.

QR 13.0a 13.5a 12.0a 12.8A 0.18

QL 16.5b 16.8b 16.5b 16.6B 0.42
2 QR 32.5a 33.2a 32.8a 34.0a 33.1C 0.28
2 QL 42.5b 43.0b 40.3b 41.0b 41.70 0.78
3 QR 108.7a 108.5a 108.5a 109.5a 108.5a 108.7E 232
3 QL 129.6b 126.3ab 121.5ab 121.1ab 117.7ab 123.2F 3.27
4 QR 144.8a 146.6a 144.8a 145.2a 145.4a 145.5a 145.4G 0.72
4 QL 172.3d 165.0cd 162.0bc 161.0bc 160.1bc 157.5b 163.0H 1.06

'"-"·...,Different letters indicate statistically significant differences (ANOVA; LSD 0.95 test). Lowercase letters indicate differences between means
for larvae of the same age in queenrisht (QR) and queenless (QL) colonies (6 means for 1-<hy-old larvae, and 12 means for 4-<hy-old larvae).
Capital letters indicate differences between overall means for all days, for larvae of all fo<r ages in queenright and queenless colonies (8 means).
120 Woke

TABLE 2. Mean amount of food (mg) in 10 cells with larvae of different age on subsequent
days after dequeening, in three queenright (QR) and three queenless (QL) colonies in spring
(17 May to 19 June). Each number for a particular day presents a mean of 15 repetitions of
food collected from 150 larvae.

Age of QR Oars after de9ueening Overall

larvae orQL 1 2 3 4 5 6 7 mean
(days) ± s.e
QR 27.6a 329ab 35.3bc 32.5ab 32.1A 0.74
QL 55.5d 41.3c 38.1bc 40.7c 43.9B 1.78
2 QR 48.1a 58.0b 67.2cd 65.5bc 71.5cde 62.1C 1.56
2 QL 88.7gf 79.9f 78.9e 71.7cdef 75.1def 78.9D 1.49
3 QR 79.3a 93.2b 98.1bc 97.2bc 109.1c 113.5c 98.4E 2.56
3 QL 126.9d 108.9c 104.7bc 106.9bc 104.1bc 100.4bc 108.7F 2.07
4 QR 104.7a 128.0abc 130.5abc 114.7ab 126.5abc 142.9bcd 130.5abc 126.3G 3.16
4 QL 164.0d 149.5cd 143.5bcd 144.2bcd 149.3cd 1225abc 137.5bcd 144.0H 4.89

A-''-'1:>ifferent letters indicate statistically slgniflcallt differences (ANOVA; LSD 0.95 test). Lowercase letters indicate d~ferences between m~Bns for
larwe of the same age In queenright (QR) and queenless (QL) colonies (8 means for 1-day-old larwe, and 14 means for 4-day-old larwe). Capital
letters indicate differences between overall means for aD days, for larwe of all four ages In queenright and queenless colonies (8 m~Bns).

spring was much higher than in the autumn in both
queenless and queenright colonies.
Larvae in queenright colonies received increasing
amounts of food each day as time passed. This differed
from the autumn results, when the food supply was
similar in successive days. The gradual increase in food
supply observed in the spring is probably due to
improving nectar and pollen flows, and increases in the
nurse bee population.
On consecutive days after division, larvae of the same
age received more food in queen less than in queenright
colonies, although, this was significant only for the first
two or three days after dequeening. These results dif-
fered from those obtained in the autumn, when signif-
icant differences also were found several days later.
However, the tendency to supply larger amounts of lar-
val food in queen less colonies in later days was similar
in both seasons.
Although the amount of food provided was higher in
queenless than in queenright colonies, a tendency to
decrease the food supplied to larvae in queenless
colonies as time passed was observed. The difference
in decreased food supply was significant in several
instances. Significant interaction (QR!QLx days of col-
lection) was found for larvae of all four ages (P <
0.0001-0.0175). Consequently, the surplus of larval
food in queen less colonies over that in queen right units
decreased as time passed.
Figure 2 shows that the relative food supply in queen-
less colonies was the highest for 1-day-old larvae
(137%). The relative amount decreased for older larvae
reaching the lowest level for 4-day-old ones (114%).
This is opposite to the total amount, which was high-
er in cells with older larvae (table 2).
Over consecutive days, the largest increase in ratios of
food provided was recorded the day after dequeening.
Larvae of all ages present in queenless colonies
received 175% of the food supplied to larvae in queen-
right colonies. The largest supply (201%) was observed
with 1-day-old larvae, the day after dequeening. This is
opposite to the absolute amount of food supplied to
larvae of different ages (table 2). As time after dequeen-
ing passed, the relative amount offood in cells with lar-
vae of all ages decreased to 105% the seventh day.
Thus, with passing of time, the amount of food pro-
vided to larvae in queenless colonies approached that
present in the queen right ones, after one week.
Relationship between amounts of food supplied
to larvae in queenright and queenless colonies
in autumn Table 3 shows that in autumn, the mean
amount offood supplied to larvae was higher in queen-
less than in queen right colonies. The additional amount
of food supplied to older larvae, (2--4-days-old), over
that given to 1-day-old larvae was higher in queen less
(25.1-146.4 mg) than in queenright colonies
(20.3-132.6 mg), (table 3, x,-x,). However, 3- and 4-
day-old larvae received as much as 8.5 and 11.4 times
more food than the 1-day-old larvae in queen-right
colonies, while only 7.4 and 9.8 times more in queen-
less ones, respectively (table 3, x,: x,). Thus, the mag-
nitude of the increased amount of food supplied to
older larvae was lower in queenless colonies. This
lower ratio is probably the result oft he greater amount
Larval food in gueenless colonies 121

._
250

200

150
~ ~r:-
~
rl. ~ ~
r-7· _;:-
100 1- 1-
~
r--

50 1::: 1::: ;:::: ~

'---
~ ,..:::: ~
'---- I--
~
0
Days 1 2 3 4 5 6 7 Mean
Mean El 175 124 111 118 106 87 105
4 days D 157 117 110 ! 126 118 86 105 114
3days o 160 117 107 110 95 88 111
2 days Iilli 184 138 117 110 105 127
1 day • 201 126 108 125 137

FIG. 2. Percentage of the amount of larval food observed in brood cells in queenless colonies in
ratio to that in queenright colonies in spring (see fig. 1).

TABLE 3. Comparison of the amount of larval food ( mg) in different seasons in queen right
(QR) and queenless (QL) colonies.

Age of Mean mgfood Difference x,-x1 Magnitude x, : x1 Relation QL QR

larvae QR QL QR QL QR QL difference magnitude
(days) QL-QR QUQR

Autumn
12.8 16.6 0.0 0.0 1.0 1.0 3.8 1.30
2 33.1 41.7 20.3 25.1 26 2.5 8.6 1.26
3 108.7 123.2 95.9 106.6 8.5 7.4 14.5 1.13
4 145.4 163.0 132.6 146.4 11.4 9.8 17.6 1.12
Spring
1 32.1 43.9 0.0 0.0 1.0 1.0 11.8 1.37
2 62.1 78.9 30.0 35.0 1.9 1.8 16.1 1.27
3 98.4 108.7 66.3 64.8 3.1 2.5 10.3 1.11
4 126.3 144.0 94.2 100.1 3.9 3.3 17.7 1.14

~ Means for 1-day-old lartae.

' Means for 2-. 3- or 4-day-old lartae. respectively.
122
of food supplied to 1-day-old larvae in queen less than
in queen right colonies, i.e. 16.6 mg vs. 12.8 mg. respec-
tively.
Table 3 shows that the increased amount offood sup-
plied to larvae in queen less colonies over that in queen-
right ones, reached from 3.8 mg to 17.6 mg for larvae
1--4 days old, (QL-QR), respectively. The amount of
food supplied to larvae 1--4 days old was from 1.30 to
1.12 times higher in queenless than in queenright
colonies (QL/QR), respectively.
Relationship between amounts of food supplied
to larvae in queenright and queenless colonies
in spring Table 3 shows that in spring, larvae received
more food in queenless than in queenright colonies.
Larvae 2 to 4 days old received 30.0-94.2 mg more
food than the 1-day-old larvae in queenright colonies,
and as much as 35.0-100.1 mg more in queen less ones,
respectively (table 3, x.~,). Thus, additional food given
to older larvae was higher in queen less than in queen-
right colonies. The 2--4-day-old larvae received 1.9-3.9
times more food than the 1-day-old in queenright
colonies, while only 1.8 to 3.3 times more in queen less
ones, respectively (table 3, x. : x,). Thus, the addition-
al amount of food given to older larvae was relatively
lower in queenless than in queenright colonies. This
relatively lower increase of food supplied to older lar-
vae was probably caused by the much greater amount
of food supplied to 1-day-old larvae in queen less than
in queenright colonies (43.9 mg vs. 321 mg). Larvae 1
to 4 days old received from 11.8 mg to 17.7 mg more
food in queen less than in queenright colonies (table 3
QL-QR). The amount of food supplied to those lar-
vae was 1.37-1.14 times higher in queenless than in
queenright colonies (table 3, QLIQR).
Comparison of amounts of food supplied to lar-
vae in autumn and spring Great differences
between the autumn and spring results were noted in
both queenright and queen less colonies. Larvae 1-day-
old received about 2.5 times more food in the spring
than in the autumn in both queenright and queenless
colonies, i.e. 321 mg vs. 12.8 mg and 43.9 mg vs. 16.6
mg. respectively (table 3, x). In the autumn, 2--4-day-
old larvae received 2.6 to 11.4 times more food than
the 1-day-old larvae in queen right colonies, and 25 to
9.8 in queenless units, respectively (table 3, x. : x,).
However, in the spring, those larvae received only 1.9
to 3.9 times more food in queen right colonies, and 1.8
to 3.3 times more in queenless units, respectively. This
relatively lower increase in food for older larvae over
that given to 1-day-old larvae in the spring may be the
result of the 2.5 times greater food supply for the
youngest larvae in the spring than in autumn.
In autumn, nurse bees fed to 1-4-day-old larvae
3.8-17.6 mg more food in queen less than in queen right
colonies (table 3, QL-QR). In the spring they fed
11.8-17.7 mg more food to larvae in queenless units.
Thus, the surplus offood given to 1-2-day-old larvae
in queenless colonies in spring was higher than in
Woke
autumn. However, the surplus of food given to 4-day-
old larvae in queen less colonies was almost identical in
both seasons.
In the autumn, 1--4-day-old larvae received 1.30-1.12
times more food in queenless, than in queenright
colonies, vs. 1.37-1.14 times more in the spring (table
3, QUQR). Thus, the absolute amounts of food fed to
the larvae in autumn and spring were seasonally differ-
ent. However, the increased ratios of food fed to var-
ious aged larvae in queenless colonies over that in
queen right units remained constant over the seasons.
Time period between dequeening of colonies
and construction of emergency queen cells
Observations over several years often revealed that the
first emergency queen cells were present in colonies
the day after dequeening. However, emergency queen
cells were not found in all colonies the day after
dequeening, and in some cases even after 2 days. This
was observed more frequently in the autumn, than in
the full season. Later, additional emergency queen cells
were constructed 2 and 3 days after dequeening.
Unfortunately, the number of cases was not recorded.
DISCUSSION
Since egg developmental time is 3 days. 1-day-old lar-
vae should be present in queen less colonies only for
up to 3 days. However, in the spring, 1-day-old larvae
were found in queen less colonies on the fourth day
after dequeening, and those 2-, 3- and 4-day-old larvae,
also found on the fifth, sixth and seventh day. The
amount of larval food fed to those larvae in the last
days was similar to that found in the previous days
(table 2). This indicates that older larvae were not mys-
tically selected as younger ones. The presence of young
larvae of a particular age for longer periods may be
explained by findings the of Milne et a/. (1988) and
Woyke (1998). According to them, some larvae hatch
more than 79.5 h after egg deposition.
After dequeening colonies, all larvae were fed increased
amounts offood. Four-day-old larvae were also fed
larger amounts of mixed food, although larvae of this
age that receive this kind of food are not selected by
workers for emergency queen cell construction.
The increased amount of larval food in queenless
colonies is not the result of different ratios of the nu m-
ber of workers to the number of larvae in queenright
and queenless colonies. During the first 3 days after
dequeening, that ratio was similar in both types of
colonies. Despite this equality, there was more larval
food in queen less colonies. During the last 3 days that
open brood was present in queenless colonies, 1-, 2-
and 3-day-old larvae were successively absent. Thus,
the ratio of the number of workers to that of the lar-
vae increased. Despite this, the amount of larval food
in queenless colonies decreased as time passed. Thus,
the changes in the amount of larval food in queen less
colonies are caused by the absence of the queen and
Larval food in gueenless colonies 123

the lapse of time after dequeening. The absence of

queen substance probably releases behavioural changes
during the days after dequeening. Later, the bees grad-
ually become accustomed to this situation.
Kuwabara (1947) observed feeding activity of nurse
bees toward 50 larvae of the same age in a queen less
colony. The number of larval feedings shortly after
hatching was four times higher in the queen less than in
a queenright colony (20 vs. 5 times/h). As larval age
increased up to 3.5 days, the number of feedings in
queenless colonies decreased to that observed in the
queenright colonies. A relationship between the
amount of larval food in brood cells presented in this
paper and the number of feeds demonstrated by
Kuwabara is apparent. However, results presented in
this paper indicate that the decreased number of feeds
3 days after dequeening is not the result of aging of lar-
vae. The decreased feeding occurs as time after
dequeening passed, independent of the age of larvae.
In some colonies, emergency queen cells were not pre-
sent the day after dequeening. Many additional emer-
gency queen cells were started 2 or 3 days after
dequeening. However, the increased amount of food
supplied was noted already the day after dequeening.
This supports the view that all larvae are supplied with
larger amounts of food after dequeening and later
emergency queen cells are constructed over some lar-
vae. This means that nurse bees in queen less colonies
select larvae, which are already provisioned with a larg-
er amount of food, for queen rearing. The increased
food supply to larvae in queenless colonies is clearly
visible without making any measurements. Thus, the
beekeeper can suspect the loss of a queen before the
construction of emergency queen cells starts.

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GONTARSKI, H (1956) Der Nachsclv.fungsinstinkt beim Blenenvolk.
lnsectes Sociaux2(2): 347-349.
KUWABARA, M (1947) Ueber die Resulatlon im welsellosen Volke der
Honlsbiene (A pis me/lifica), besonders die Bestimmung des neuen
Weisels. journal of the Faculty of Saence, Hokkatdo Umversity, VI
Zoology 9: 359-381.
NELSON,] A, STURTEVANT. A P, UNEBURG, B (1924) Grov.th and feed-
ing ofhone)t!ee lame. United States Department of Asricult~re,
Department Bulletin No. 1222; 37 pp.
MILNE, C P; PHWPS,J P; KRELL, PJ (1988) A photomicrographic study
of worker honeybee embryogenesis. journal of ApiCultural
Research 27(2): 69-83.
WOYKE, J (1998) Size charwe of Apis me/l(era eggs do.rtng incui>atlon peri-
odjournalofAplwltural Research 37(4): 239-246.