*3.3.1 Larva transfer*

RJ is a functional food containing various physiologically active substances. It is therefore the basic principle in RJ production to harvest RJ with high content of functional components and appropriately ensure that the physiological activities are maintained. In RJ production using artificial queen cell cups, the timing of larva transfer to queen cell cups and the timing of harvest are important.

### *Kikuji Yamaguchi Principles of Natural Beekeeping: A Novel Bio-Method of Natural Beekeeping… DOI: http://dx.doi.org/10.5772/intechopen.89647*

The influence on RJ production of timing of larva transfer to queen cups was studied extensively to find that it is preferable to transfer second-instar worker bee larvae (early second-instar, larva size: about 1 mm). First-instar larvae are too small and too soft making the larva-transferring operations difficult. The larvae grow quickly, and a surprising increase of body size is achieved in only 1 day.

In the case of early second-instar larvae, the amount of RJ ingested after larva transfer is not so great, and RJ rich in effective components can be harvested. Also, the larva transfer must be performed in a tent to protect it as much as possible from ultraviolet light. The larvae are quickly transferred into the artificial queen cell cups using a transferring tool called a "larva-transferring needle" while taking care not to hurt the larvae (**Figure 6**).

#### *3.3.2 Number of artificial queen cell cups*

It is well known that RJ is stored in the queen cell cups where queen bees are reared. In ordinal beekeeping, the number of artificial queen cell cups used is approximately 200–250. On the other hand in the Natural Beekeeping (KYAMENABEE), the maximum number of artificial queen cell cups is limited to 100. It is well known that RJ is secreted by young worker bees (aged 3–12 days after becoming mature insects). However, the number of such worker bees is limited accounting for about 20% of the entire colony. Large amounts of pollen and honey are necessary for the worker bees to secrete RJ. Young worker bees eat these and secrete RJ at full power into queen bees in the artificial queen cell cups for the larvae growing.

Not a few beekeepers who cherish honeybees hate the production of RJ because it impairs honeybees and reduces the size of colony. Young worker bees will soon become foraging bees to collect flower nectar and pollen and complete their lifespan lasting 1 month. However, when they overworked to secrete too much RJ, they die within 21–30 days. The colony is gradually weakened if the number of such worker bees increases.

#### **Figure 6.**

when the apiary is located in a mountain area, apicultural products are generally collected at a processing plant located in an urban area for filtration and packaging. It is problematic to leave the unfiltered apicultural products for a long time. In the meantime, the products become less active due to the impurities contained therein. Honey may be fermented when the Brix degree is low, while RJ shows signs of

*Filtration of harvested RJ at apiary. The harvested fresh RJ is immediately filtered under a tent avoiding direct*

The apicultural products carried into the processing plant are once frozen, thawed for filtration and mixing performed in turn and then frozen again. The

The principle of natural beekeeping (KYAMENABEE) is to produce apicultural products using healthy colonies. It is therefore necessary to keep the bee hives clean all the time and protect the colonies from diseases by performing cleaning and fumigation. It is important to avoid artificial mating which leads to the weakening of colonies and antibiotics should not be used to prevent diseases and infections. Methods for improvement of the health of colonies should be adopted before using

RJ is a functional food containing various physiologically active substances. It is therefore the basic principle in RJ production to harvest RJ with high content of functional components and appropriately ensure that the physiological activities are maintained. In RJ production using artificial queen cell cups, the timing of larva

transfer to queen cell cups and the timing of harvest are important.

repeated freeze/thaw procedures cause quality to deteriorate.

*sunlight to remove foreign matters such as dead bees, hive scum and dust.*

*Modern Beekeeping - Bases for Sustainable Production*

denaturation.

**Figure 5.**

*3.2.9 Antibiotics*

antibiotics.

**126**

**3.3 RJ production**

*3.3.1 Larva transfer*

*Transferring of larvae. The larvae are quickly transferred into the artificial queen cell cups using a transferring tool called "larva-transferring needle" paying attention not to hurt the larvae, but this operation needs considerable mastery.*

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, demonstrating that such RJ is of low nutritive value and is inactive.

been manufacturing RJ since 1993 in a declared un-contaminated area of Chinese highland over 3000 m (3200–3600 m) above sea level (Qing Hai Haibei Menyuan).

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

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

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

*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*

*3.3.5 Colony control—Rotation of colonies*

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

queen cell.

**Figure 9.**

*every 4 days.*

**129**
