8 THE BACTEEIA OF THE APIAEY. 



comes in loaded with pollen and honey. After the victim alights on 

 the entrance board, by the aid of a pair of forceps, before she disap- 

 pears within, one can easily lodge her safely in a petri dish. It is, 

 however, an advantage to study the young adult bees as well as the 

 older ones, and if young ones are desired they may be taken from 

 the combs or from the front of the hive, near the entrance. 



Obtaining Cultures. 



(a) From combs. — With sterile forceps small pieces of the comb 

 are put directly into gelatin or agar for plates or incubated in bouil- 

 lon for 24 hours and then plated. Growing in bouillon and plat- 

 ing on gelatin is usually preferable. 



{h) From pollen. — The same technique is used as for combs, but 

 the direct inoculation of gelatin tubes for plates is generally pre- 

 ferable. 



(c) From honey. — With sterile loops honey is taken from uncapped 

 and capped cells. The caps are removed with sterile forceps and the 

 honey is plated directly on gelatin or agar. Bouillon tubes are in- 

 oculated also with varying quantities of the honey. 



{d) From larvm. — The larva is carefully removed to a sterile dish, 

 and with sterile scissors the body is opened and the contents plated 

 directly, or bouillon cultures are first made and later plated, if a 

 growth appears. 



(e) From parts of the adult hee. — In studying the adult bee, a 

 small piece of blotting paper wet with chloroform is slipt under 

 the cover of the petri dish in which the insects have been placed, and 

 in a short time the bees are under the influence of the anesthetic. 

 Then with sterile scissors a leg, a wing, the head, the thorax, or the 

 abdomen, the intestine being removed, is placed in bouillon and, after 

 24 hours incubation, plated, preferably on gelatin. 



When it is desired to make a study of the bacteria of the intestine, 

 the intestinal tract is removed and studied as follows: The bee is 

 flamed and held in sterile forceps. With another sterile pair of for- 

 ceps the tip. of the abdomen is seized and, by pulling gently, the tip 

 and the entire intestine are easily removed. This can then be plated 

 directly. If gelatin, which is preferable, is used, the intestine itself 

 must not be left in the gelatin or the medium will become liquefied 

 by the presence of the tissue. If one desires to obtain cultures of the 

 anaerobe, which is quite common in the intestine, it is most easily 

 obtained in pure culture by the use of the deep glucose agar (Liborius's 

 method). Cover glass preparations made direct from the walls of 

 the intestine or its contents give one some idea of the great number of 

 bacteria frequently present. 



