*3.3.5 Colony control—Rotation of colonies*

Additionally, when excess number of artificial queen cell cups is used, the quality of RJ thus harvested markedly deteriorates, and the stored RJ becomes watery and exhibits low viscosity, and the quality may be markedly deteriorated. When such RJ is given to larvae hatching from fertilized eggs, they cannot become decent queen bees and most of them become to grow up to worker bees, demon-

RJ should be harvested 48 h after larva transfer to artificial queen cell cups. This is because the RJ harvested 48 h after larva transfer is very active although the amount is smaller than the amount harvested after 72 h. As described below (Section 3.2.1), the content of 10-HDA in RJ is significantly higher in the harvest after 48 h than that after 72 h. The various effects on humans and experimental animals are also known to be more excellent in the former harvest. In the case of 72 h harvest, the operation efficiency is higher since a larger amount of RJ is pooled in each queen cell cup and the operation can be performed once every 3 days, but such the RJ harvested is far

In the Natural Beekeeping, RJ is manufactured in single bee forage of rape blossoms. The RJ obtained from rape blossoms is highly active. In particular, the pollen of rape blossoms is active and strengthens the colonies after wintering.

On the other hand, the bee forage is not specified in ordinal beekeeping, and RJ is manufactured even in the absence of bee forage by feeding with sugar water and artificial pollen. The rape blossoms growing in the flat plains of mainland China can no longer be used due to contamination from agrochemicals, and my group has

*Harvesting of RJ at 48 h (A) and 72 h (B) after larva transfer. Accumulation of RJ is much greater in the cup*

*72 h after larva transfer than that 48 h after larva transfer.*

*Collection of RJ from the artificial queen cell cup.*

strating that such RJ is of low nutritive value and is inactive.

*Modern Beekeeping - Bases for Sustainable Production*

less active compared to that after 48 h harvest (**Figures 7** and **8**).

*3.3.3 Time before harvest*

*3.3.4 Bee forage*

**Figure 7.**

**Figure 8.**

**128**

Originally, the mass production of RJ using artificial queen cell cups became feasible based on the biological and ecological properties of honeybees. Only one queen bee in the hive keeps laying 2000–3000 eggs per day. At first glance, the queen bee appears to be playing the central role in the colony, but the colony is actually controlled by the worker bees. Neither the queen bee nor the drone bees can live unless fed by the worker bees, since they have no habit of procuring foods by themselves. Since the worker bees have the feeding rights, in the absence of queen bee after isolating the queen bee from the hive, the worker bees rush towards rearing a new queen bee. When the worker bees find a third-instar or younger larva in a hive cell, they break the cell by eating, enlarge the cell to a queen bee-rearing cell cup and prepare a temporary queen cell cup (this is referred to as an "emergency queen cell"), and thereafter they start secreting RJ into the emergency queen cell. The larva that up until this point was going to grow as a worker bee is given RJ due to a policy change, and this larva ultimately emerges as a queen bee. In RJ production, utilizing the habit of honeybees to change an abnormal state without a queen bee into a normal state with a queen bee, the worker bees are made to secrete RJ by introducing artificial queen cell cups into the hive in place of the emergency queen cell.

In spite of the industrial policy for mass production of RJ, it must not be forgotten that honeybees are living organisms. Furthermore, they are extremely delicate living organisms due to their high and sophisticated capability and sociality. In order to avoid weakening the colony by reducing its disease resistance due to overloading the honeybees, I have adopted the bee colony rotation system in harvesting RJ. When harvesting, only two queen cell cup frames are inserted and only 60–100 artificial plastic artificial queen cell cups are put into each queen cell cup frame. In this way, it was confirmed that the colony with sufficient feeding becomes powerful: 100 mg of RJ can be harvested from one queen cell cup in 24 h and 300–480 mg in 48 h. This is performed in a rotation system (**Figure 9**). The

#### **Figure 9.**

*Rotation of Colony. The colonies are divided into four groups, and the colony used once for larva transfer and harvest is made to rest, and another colony is used for production. This is rotated in sequence at a rate of once every 4 days.*

colonies are divided into four groups, and the colony used once for larva transfer and harvest is made to rest, with another colony used for production. This is rotated in sequence once every 4 days. In other words, the colony made to work once will then be given a 2-day rest. Needless to say, a greater amount can be produced when all of the colonies are made to work fully, but this is a fundamentally mistaken policy. Originally, in the method of producing RJ using artificial queen cell cups, the circumstance in which worker bees prepare the emergency queen cell in response to the emergency situation after disappearance of the queen bee was created artificially. In this sense, RJ production is a significant stress on worker bees. Through the rotation of colonies, colony momentum is maintained and the lifespan of worker bees is also prolonged. Most important is that the disease resistance of the colony is increased by this rotation, leading to avoidance of the use of drugs such as antibiotics. Even when the rotation is adopted in RJ production, the colony in the resting state can be dedicated to the collection of nectar and pollen, as the queen cell cup frame is not inserted. Honeybees are insects, and it is impossible to prepare genuine RJ required by consumers unless the ecology and providence of honeybees are followed.

### *3.3.6 Filtration at apiary and temperature control*

The RJ harvested from the beehive should be subjected to primary filtration at the apiary in order to remove bee wings, etc. and is then transported to a processing plant.

It should be recommend to filter the harvested highly-active RJ immediately at the apiary and store it at 2°C under complete protection from sunlight. When RJ comes into contact with oxygen or carbon dioxide in the air or is exposed to ultraviolet light, it quickly becomes less active. It is also known that denature and turn-over of components occur immediately when RJ is left at ordinary temperature. The activities, however, can be maintained for a far long time when stored at low temperature.

the standard value. Under such circumstances, we cannot even hope to receive high-quality RJ. Therefore, in the Natural Beekeeping, the harvested RJ is filtered immediately at the apiary so as to remove foreign matters, and sufficient effort is made to avoid component changes. The RJ is stored at 2°C and cryopreserved at

*Generator and refrigerator. An electric refrigerator combined with gasoline-driven generator was supplied to*

*Kikuji Yamaguchi Principles of Natural Beekeeping: A Novel Bio-Method of Natural Beekeeping…*

Concerning the suitable temperature for storage of RJ (2°C in refrigeration and 18°C in cryopreservation), Smith had already reported in the "Bee World" journal in 1959 that "The harvested RJ must be immediately stored in a refrigerator. Oneyear storage is probably possible at 2°C. No changes were observed in royal jelly stored at 18°C for several years", and this storage method was established worldwide [23]. Thereafter, the optimal temperature for storage being 2°C was also reported in the German beekeeping journal "Archive Hule Bienenkunde", and Inoue, a Japanese beekeeping researcher [3], stated that "The opinions of the world's researchers are mostly in unison" (cited from "New technology for higher

In 1967, I supplied an electric refrigerator combined with a gasoline-driven generator to Japanese beekeepers so that five beekeepers could share one set. At that time, prevailing wisdom dictated that RJ should be stored at ordinary temperatures

under a tent, and the beekeepers resisted the introduction of refrigerators, complaining that they were "troublesome", "owner indistinguishable", "may be stolen", "have questionable security", and so on. I took the following countermeasures: the neat fluids were accommodated in plastic bags with different colors, and the manufacturing site, manufacturer, date and time of harvest, etc. were written on each bag with a magic marker. In other words, attention was paid to perfect traceability even at that point in time. This system was also introduced to the rape

blossom bee forages in Qing Hai Haibei Menyuan.

18°C after packaging. It is frozen only once on this occasion.

*beekeepers so that the temperature control was easily performed at the site.*

*DOI: http://dx.doi.org/10.5772/intechopen.89647*

yield of royal jelly").

**131**

**Figure 10.**

Therefore, in the Natural Beekeeping (KYAMENABEE), the harvested RJ is immediately filtered so as to remove foreign matter such as dead bees, hive scum and dust. Although denaturation occurs immediately when left at ordinary temperature, it is known that RJ can remain active for a fairly long time when stored as at 2°C. I recommend filtering the harvested RJ on every harvest so as to remove impurities, followed by temporary storage at 2°C under protection from sunlight and final cryopreservation below 18°C (**Figure 10**).
