MORAXELLA

Prepared by: Prem G.M

Class Roll no. 30
Exam roll no. : 227(5th sem)
 MORAXELLA
Introduction
 The genus Moraxella is a member of the family Moraxellaceae.
 It has been suggested that the genus should be divided into two subgenera,
Moraxella and Branhamella.
 The moraxellae are short, plump, gram negative rods (1.0–1.5 ×1.5–2.5 µm) that
characteristically occur in pairs (particularly those of the subgenus Branhamella).
 Some strains approach a completely coccal shape.
 They are strict aerobes, oxidative, oxidase-positive, usually catalase-positive
non-motile and do not attack carbohydrates.
 Although they will grow on non-enriched media, their growth is enhanced by the
addition of blood or serum and the optimal temperature for growth is 33–35°C.
 Most M. phenylpyruvica strains will grow on MacConkey agar, but M. bovis and M.
lacunata are unable .
 The principal animal pathogen in the group is Moraxella bovis but M. ovis and M.
equi are also associated with disease in animals.
 Morphology and Staining
 The cell wall is typical of gram-negative bacteria being composed of
lipopolysaccharide (LPS) and protein.
 The LPS of moraxellae does not contain O-repeat units many other
gram-negative microorganisms (e.g. members of the family
Enterobacteriaceae).
 The fimbrial adhesins (pili) of M. bovis are virulence determinants and
can be lost in subculture .
 Capsules maybe present on fresh isolates.
 It consists of following components:
Adhesins :
The role of adhesins, as in other microorganisms, is to allow the
bacterium expressing them to adhere to cells lining a particular niche, as
well as to the surface of the so called target cells prior to the initiation of
disease (in some cases, niche and target cells may be the same). M. bovis
produces a type 4 pilus (fimbria) that adheres to conjunctival and corneal
epithelial cells. This pilus is similar to those of Pseudomonas aeruginosa,
Neisseria gonorrhoeae, Dichelobacter nodosus, Pasteurella multocida, and
Vibriocholerae.Mutants unable to produce this adhesion are avirulent.
 Exotoxins:
The most noteworthy toxin produced by M. bovis is an RTX (repeats in
toxin so called because of the structural repeats of glycine-rich
sequences within the protein) type of cytotoxin .This cytotoxin,
sometimes referred to as “hemolysin” due to its behavior on blood
agar plates, has been termed Mbx (for M. bovis toxin). Mbx is a pore-
forming toxin with specificity for conjunctival and corneal epithelial
cells, and neutrophils. Mutants unable to produce Mbx are avirulent.

Iron Acquisition:
Because iron is an absolute growth requirement, microorganisms must
acquire this substance If they are to exist within the host. Moraxellae
acquire iron from their on-binding proteins of the host(transferrinand
lactoferrin)by expressing Tbp and Lbp (fortransferrin-and lactoferrin-
binding proteins ,respectively) on their surface. Tbp and Lbp bind their
respective proteins giving moraxellae access to iron.
Miscellaneous Products:
A number of proteins with toxic activities are produced in vitro by
M. bovis. These include complement-degrading proteases, lipases,
phosphoamidases, peptidases, and proteases. There is little
evidence showing that any of these play a role in vivo.

Growth Characteristics:
M. bovis grows best at 35 ◦C in the presence of serum and blood.No growth
occurs on MacConkey agar or anaerobically.In 48h,fresh isolates produce flat,
hemolytic ,friable colonies, about 1 mm in size, that corrode the agar and auto
agglutinate when suspended in saline.

Biochemical Activities:
M. bovis is oxidase-positive, asaccharolytic, non-fermenting, and
catalase-variable. Nitrates and urea are not digested, but proteins are
digested. Most other species of Moraxella do reduce nitrate and nitrite.
 Capsule:
The capsule plays many roles, the most important of which are
interference with phagocytosis (antiphagocytic), and protection of the
outer membrane from the deposition of membrane attack complexes
generated by activation of the complement system.

Cell Wall:
The cell wall of the members of this genus is one typical of gram-
negative bacteria (except for the absence of the O-repeat unit). The
LPS in the outer membrane is an important virulence determinant.
LPS binds to LPS binding protein (a serum protein), which transfers it
to the blood phase of CD14. The CD14–LPS complex binds to Toll-like
receptor proteins(seeChapter2)on the surface of macrophage cells,
triggering the release of pro inflammatory cytokines.
Resistance:
Resistance to physical and chemical agents is not remarkable.It is usually
susceptible to commonly used antibiotics including penicillin.Some
isolates of M.bovis are resistant to tetracyclines and tylosin.Moraxella
catarrhalisis the only species that commonly produces aβ-lactamase.

Isolation:
Lacrimal secretions should be inoculated as soon as possible after
collection on blood agar and incubated at 35°C for 48–72hours. The
inoculation of a MacConkey plate is useful to gauge the degree of
contamination by other Gram-negative bacteria, M. bovis is unable to
grow on MacConkey agar.

Pathogenesis and Pathogenicity:

Moraxella bovis, the cause of infectious bovine keratoconjunctivitis,
is the major animal pathogen in the genus .The pathogenicity of M
bovis strains depends on the production of type IV pili and a
cytotoxin.
Cont……
The cytotoxin is a haemolysin and belongs to the repeat-in-toxin family of toxins
(Angelos et al. 2001). Strains which are not haemolytic or lack pili are apathogenic
for cattle.In addition to the pili, which have a major role in attachment of the
organism, filamentous-haemagglutinin-like proteins may be important in the
infectivity of the organism (Kakuda et al. 2006). Other enzymes which may play a
role in tissue destruction include fibrinolysins, proteases and
phospholipases.Acquisition of iron through transferrin and lactoferrin-binding
proteins may contribute to virulence also. Predisposing environmental factors are
implicated in infectious bovine keratoconjunctivitis.These include irritation to the
eyes by ultra-violet light (in sunlight), dust, long vegetation and flies. The
incidence of the disease is highest in the summer months. Concurrent infections
by such agents as Mycoplasma bovoculi, Listeria monocytogenes, Chlamydophila
pecorum, bovine herpesvirus1, bovine adenoviruses and the nematode Thelazia
species may complicate the disease. Young animals under two years of age are
most commonly affected and in a typical outbreak of disease about 60% of the
young cattle maybe affected while only about 20% of the older animals are
affected. The early signs of infectious bovine keratoconjunctivitis are
acrimation,blepharospasm and conjunctivitis. Later an ulcer develops on the
cornea. Corneal opacity and oedema surround the ulcer and in severe cases
vascularization of the cornea occurs from the limb us to the ulcer. The corneal
opacity then involves the entire cornea. In the healing stage, granulation tissue
forms on the ulcer floor and a characteristic red cone of granulation tissue will
project from the cornea. The granulation tissue and the ulcer itself will eventually
regress leaving a white corneal scar. The scar may or may not be permanent.
Laboratory Diagnosis:
The agent maybe demonstrated in smears of exudate ,most convincingly
by immunofluorescence (by using the antibody specific to M. bovis
antigens). Exudate is cultured on blood agar, and Moraxella is identified
by colonial characteristics, oxidase activity, hemolysis, proteolysis, and
failure to ferment carbohydrates. Specific fluorescent antibody
conjugates can be applied directly to suspect colonies on plates for
identification even of dissociant colonies (epifluorescence). Polymerase
chain reaction-based assays utilizing M.bovis-specific primers are
available for detection and identification.