EXTRACTS FROM THE BERGEY'S MANUAL OF SYSTEMATIC BACTERIOLOGY

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Family Anaplasmataceae

GENUS ANAPLASMA
(Bergey 's Manual of Systematic Bacteriology)

Small, often pleomorphic, coccoid to ellipsoidal cells 0.3 – 0.4 µm in diameter found in cytoplasmic vacuoles in
mammalian host cells, often in inclusion bodies (morulae) in mature or immature hematopoietic cells, in
peripheral blood, or in tissues, usually organs containing mononuclear phagocytes (spleen, liver, bone marrow)
of mammalian hosts. Morulae are suspended in a non-fibrillar matrix. Two morphologic forms: larger reticulate
cells and smaller “dense core” forms. In blood smears stained with Romanowsky methods, the organisms
appear as dense, homogeneous, bluish-purple, round inclusions 0.3 – 2.5 µm in diameter. Gram negative. Non-
motile. No spores or resistant stages. Ticks are the only known biological vectors; mechanical vectors include
biting flies or other fomites. The organism can be propagated in tick spp. Some Anaplasmas can be propagated
in tick cell lines or mammalian cells of hematopoietic origin. Cause disease in canids, humans, and ruminants.
Variably pathogenic in cattle, goats, sheep, deer, horses and rodents.

The genus Anaplasma includes A. phagocytophilum, infecting numerous species of mammals including
humans; the ruminant pathogens A. marginale, A. centrale, A. ovis, and A. bovis; and the canine pathogen A.
platys.

Type species: Anaplasma marginale

Cell morphology: Anaplasma species stain bluish-purple with Romanowsky methods. They occur in
membrane-bound vacuoles in the cytoplasm of cells of hematopoietic origin, forming inclusions that contain
variable numbers of organisms. Various species infect predominantly erythrocytes, neutrophils, or
mononuclear phagocytes in vivo. The individual organisms are approximately 0.3 µ in diameter but may vary
considerably in size and shape. Inclusions (morulae) range in size up to 4.0 µm in diameter. The organisms are
weakly Gram-negative. Anaplasmas are best stained with Romanowsky stains, where the organisms develop a
dark blue appearance as opposed to the violet colour of the eukaryotic cell nucleus. The organisms stain poorly
with the Giménez method but easily with acridine orange.

GENUS EHRLICHIA
(Bergey 's Manual of Systematic Bacteriology)

Coccoid to ellipsoidal cells. Found in cytoplasmic vacuoles in endothelial or hematopoietic mammalian host
cells, including macrophages, monocytes, neutrophils, and cells of the bone marrow, liver, spleen, or lymph
nodes. Often occur in clusters (morulae). Pleomorphic; occur as reticulate cells and “dense core forms” with
condensed cytoplasm. Gram negative. Non-motile. Some species grow in cultured tick cells or in cultured

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mammalian monocytes, macrophages, or endothelial cells. Cause disease in ruminants, canids, rodents, and
humans. Ticks are the primary vectors and hosts.

The genus Ehrlichia includes five species. Three species (E. canis, E. ewingii, and E. chaffeensis) cause infection
in dogs and humans; E. muris is a murine pathogen, and E. ruminantium causes heartwater disease in domestic
ruminants.

Type species: Ehrlichia canis

Cell morphology: Ehrlichia occur in membrane-bound vacuoles in the cytoplasm of host cells of hematopoietic
origin and some species in endothelial cells. The bacteria form inclusions (morulae) that contain variable
numbers of organisms. The individual organisms are approximately 0.5 µm in diameter, but size may vary
considerably. The morulae range in size up to 4.0 µm in diameter. The organisms are weakly Gram negative.
Ehrlichia are best stained with Romanowsky stains, where the organisms develop a dark blue appearance as
opposed to the violet colour of the host cell nucleus. Organisms stain poorly with the Giménez method, but
easily with acridine orange.

GENUS NEORICKETTSIA
(Bergey 's Manual of Systematic Bacteriology)

Coccoid or pleomorphic cells that reside in cytoplasmic vacuoles within monocytes and macrophages of dogs,
horses, bats, and humans. Small (0.2 – 0.4 µm) electron dense forms and relatively large (0.8 – 1.5 µm) lighter
forms may occur. Tissues of adult trematode (fluke) vectors and all other fluke stages (eggs, miracidia, rediae,
sporocysts, cercariae and metacercariae) produce infection when injected into susceptible hosts, indicating
transovarial transmission in the vector. Transstadial transmission also occurs. Gram negative. Non-motile.
Most species are cultivable in peripheral blood monocytes, myelomonocytic cell lines, or promyelocytic cell
lines. Some species are pathogenic to laboratory mice. Etiologic agents of diseases of dogs and other canids,
horses, and humans.

Type species: Neorickettsia helminthoeca

Cell morphology: Neorickettsia species are found in the cytoplasm of monocytes in the blood and in
macrophages of lymphoid tissues of infected Canidae, humans, bats and horses. In Romanowsky-stained
preparations the organisms are dark blue to purple. Individual round forms are most common, but small
clusters (micro-colonies) of bacteria in inclusions (morulae) are also present. These may be mistaken for large
bacteria. The morulae are less compact and are smaller than those of Ehrlichia spp. in DH88 cells.

GENUS WOLBACHIA
(Bergey 's Manual of Systematic Bacteriology)

Pleomorphic bacteria that appear as small rods (0.5 – 1.3 µm in length) and coccoid forms (0.25 – 1.0 µm);
large forms (1.0 – 1.8 µm in diameter) may be observed. Grow in vacuoles of host cells. Do not grow outside of
cells. Associated with arthropods and filarial nematodes. Responsible for reproductive alterations in

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arthropods. With few exceptions, not pathogenic for the host. Stains associated with nematodes responsible
for indirect pathogenesis in humans. Susceptible to doxycycline and rifampin demonstrated in vitro and in vivo.

Type species: Wolbachia pipientis

Cell morphology: Wolbachia pipientis is not visible by Gram staining in spite of its Gram-negative cell wall
structure. The organisms can be stained using conventional Giemsa’s stain or rapid staining such as Diff-Quick.
With the Giménez stain, the bacteria can be visualized, but they appear as dark blue structures within a blue-
green cytoplasm and not as pink-red structures as do rickettsiae.

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Family Rickettsiaceae

GENUS RICKETTSIA
(Bergey 's Manual of Systematic Bacteriology)

Short, often paired rods, 0.3–0.5 x 0.8-2.0 µm. The rickettsial envelope has a typical Gram-negative structure
with a bilayer inner membrane, a peptidoglycan layer, and a bilayer outer membrane. The cells are often
surrounded by a protein microcapsular layer and slime layer. Rickettsiae retain basic fuchsin when stained by
the method of Giménez. The organisms are obligately intracellular and reside free in the cytoplasm of the
eukaryotic host cell, where they divide by binary fission. Rickettsiae of the spotted fever group (SFG) may also
reside in the nucleus of the eukaryotic host cells. Rickettsiae are closely associated with arthropods (ticks,
mites, fleas, lice and other insects) for their maintenance in nature. Their natural cycle usually involves both a
vertebrate and invertebrate host. For some, the arthropod host is both a reservoir and a vector. Transovarian
transmission of the agent from the infected female to the next generation is the essential mechanism for the
maintenance of many species. Rickettsiae are etiological agents of typhus and spotted fevers in humans. There
are 21 recognized species.

Rickettsia species are divided into three groups based on phylogeny. The typhus group (TG) includes R.
prowazekii and R. typhi. The spotted fever group (SFG) includes R. aeschlimannii, R. africae, R. akari, R.
australis, R. conorii, R. felis, R. helvetica, R. honei, R. japonica, R. massiliae, R. montanensis, R. parkeri, R.
peacockii, R. rhipicephali, R. ricketsii, R. sibirica and R. slovaca. The third group, the ancestral group, includes R.
belli, R. canadensis and the AB male-killing bacterium; this group represents early divergent lineages within the
genus.

Type species: Rickettsia prowazekii

Cell structure: Although rickettsiae are relatively small bacteria, they closely resemble other Gram-negative
bacteria. In smears from yolk sac, tissue, or cell culture, rickettsiae are best visualized by the Giménez stain. By
this procedure, Rickettsiae stain bright red with basic fuchsin, while the background is decolorized and stains
pale greenish blue with the malachite green counterstain.

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 GENUS ORIENTIA
(Bergey 's Manual of Systematic Bacteriology)

Short rods 0.5–0.8 x 1.2-3.0 µm. Obligately intracellular. Grow free in the cytosol of the host cell; rarely invade
the nucleus. Bacterial cells are released surrounded by host cell membrane. Cell walls lack lipopolysaccharide
and peptidoglycan. The outer leaflet of the cell wall is considerably thicker than the inner leaflet, unlike the
wall of rickettsiae. Maintained by transovarian transmission in its various trombiculid mite hosts. Etiological
agents of scrub typhus in humans. Transmitted to humans by mite larvae. Found in Asia and Australia.

Type species: Orientia tsutsugamushi

Cell structure: Orientia tsutsugamushi is stained by a modification of the Giménez procedure used for the
other rickettsiae; this modified procedure require preliminary destaining with ferric nitate and counterstaining
with fast green. This species is also satisfactorily stained by Giemsa stain after Carnoy’s fixation.

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