Septicemia
Pseudomonas aeruginosa (= Pseudomonas apiseptica) is the bacterium that causes septicemia in honey bees. This disease results in the destruction of connective tissues of the thorax, legs, wings, and antennae. Consequently, the affected bees fall apart when handled. Dead or dying bees may also have a putrid odor.

Pseudomonas aeruginosa rods measure 0.5- 0.8 by 1.5- 3.0 µm. They are Gram-negative and occur singly, in pairs, or in short chains. A bacterial smear and Gram stain can be easily prepared after removing a wing from the thorax and dipping the wing base in a drop of water on a microscope slide. To isolate this organism, streak the base of a wing across Difco Pseudomonas isolation agar or Pseudomonas Agar F. The optimum temperature for growth is 37?C. Pseudomonas aeruginosa in culture is characterized by the excretion of diffusible yellow- green pigments that fluoresce in ultraviolet light (wavelength below 260 nm).

Septicemia disease can also be diagnosed by reproducing the disease symptoms in healthy caged bees. This is accomplished by preparing a water extract (macerate the equivalent of one suspect bee per mL of water) and inoculating healthy bees through the thorax or dipping them in the water extract. Bees with septicemia die within 24 hours and exhibit the typical odor and the "break apart" symptom after approximately 48 hours.

Spiroplasmosis
Spiroplasma species is the bacterium that causes spiroplasmosis. Spiroplasma is a helical, motile, cell-wall-free prokaryote that is found in the hemolymph of infected adult honey bees. The organism is a tiny, coiled, and sometimes branched filament 0.7-1.2 µm in diameter (fig. 12). Its length increases with age and ranges from 2 to >10 µm (Clark 1977, 1978a).

Spiroplasma can be seen in the hemolymph using the oil immersion objective of a phase-contrast microscope. Hemolymph can be taken from adult bees by puncturing the intersegmental membrane directly behind the first coxae with a fine capillary tube made from the tip of a Pasteur pipet.

This organism can be cultured in standard mycoplasma broth medium (GIBCO) and in Singh's mosquito tissue culture medium with 20% fetal calf serum.