AN ASSIGNMENT

SUBMITTED BY: MEHAK WAHAB

SUBMITTED TO: MA’AM HIRA MUSHTAQ
SUBJECT: MEDICAL MICROBIOLOGY
CLASS: MICROBIOLOGY, 4TH
SEMESTER, MORNING
ROLL NO: 44
TOPIC: PROTEUS SPP.
DATE: 20-06-2022

CENTRE OF BIOTECHNOLOGY AND

MICROBIOLOGY
 PROTEUS SPP.
PROTEUS:
Proteus is a member of the Enterobacteriaceae family. The genus of Proteus
consists of motile, aerobic and facultatively anaerobic, Gram-negative rods.
Proteus is a member of the tribe Proteeae, which also includes Morganella and
Providencia.

CHARACTERISTICS:
• Proteus spp. are Gram-negative aerobic bacteria.
• Their size is 0.4–0.8 μm in diameter and 1.0–3.0 μm in length.
• Proteus spp. are motile.
• Rod shape
• They are non-capsulated.
• They possess peritrichous flagella.
• Non-spore forming
• Non-lactose fermenting

SPECIES OF GENUS PROTEUS:

The genus Proteus currently consists of five named species:
P. mirabilis
P. vulgaris
P. penneri
P. myxofaciens
P. hauseri
Three unnamed genomospecies: Proteus genomospecies 4, 5, 6. However, a
recent study indicated that P. myxofaciens may represent a separate genus with
low similarity to tribe Proteeae, and it has been suggested that this organism be
renamed Cosenzaea myxofaciens.

OCCURRENCE:
Proteus spp are most commonly found in the human intestinal tract as a part of
normal human intestinal flora but it becomes pathogenic when it leaves the
intestinal tract especially when it become in contact with urea in the urinary tract.

MAIN SPECIES OF MEDICAL IMPORATNCE:

• P.mirabilis
• P.vulgaris
P.mirabilis causes urinary tract infections (UIT) . P.vulgaris causes nosocomial
infections.

VIRULENCE FACTORS:
• Urease
• Hemolysins
• Motility & Swarming
• Flagella & fimbriae
• Proteinase

1. UREASE:
• Is a critical virulence determinant for colonization of the urinary tract ,stone
formation & the development of acute pyelonephritis.
• Urease –induced formation of ammonia protects the bacterial cell from the
effects of complements.
• => causes rapid hydrolysis of urea with liberation of ammonia . Thus, in urinary
tract infections with Proteus, the urine becomes alkaline ,promoting stone
formation.

2.FIMBRIAE:
• Also play a significant role in virulence for the urinary tract also have a role in
the colonization of the upper part of the urinary tract.
• Their presence correlates with the development of acute pyelonephritis.

3.HEMOLYSINS:
• Also known to be important virulence factor of proteus spp.
• Both HpmA & HlyA are cytotoxic for a wide variety of cell types , together with
urease they play an important role in cell invasion & internalization.
• Hemolysins and urease are expressed at higher levels in P. mirabilis than in
P.vulgaris and this may be the explanation for the greater virulence o.f P.mirabilis.

4.PROTEINASE:
Play a role in :
• Degrading antibacterial peptides in the innate defense system.
• Generating products like glutamine to induce invasive swarm cell.

ANTIGENIC CHARACTERISTICS:
→ The bacilli possess thermostable, ‘O’ (somatic) and thermostable ‘H’ (flagellar)
antigens, based upon which several serotypes have been recognized.
→ Certain strains of Proteus vulgaris(OX-19, OX-2 ) and proteus mirabilis (OX-K)
produce O antigens that are shared by some rickettsiae.
→ These Proteus strains are used in an agglutination test (the Weil-Felix test) for
serum antibodies produced against rickettsiae of the typhus and spotted fever
groups.

PATHOPHYSIOLOGY:
• The first step in the infectious process is adherence of the microbe to host
tissue. Fimbriae facilitate adherence and thus enhance the capacity of the
organism to produce disease.
• Fimbriae are tiny projections on the surface of the bacterium. Specific chemicals
located on the tips of pili enable organisms to attach to selected host tissue sites
(eg, urinary tract endothelium).
• The presence of these fimbriae has been demonstrated to be important for the
attachment of P mirabilis to host tissue.
• The ability of Proteus organisms to produce urease and to alkalinize the urine by
hydrolyzing urea to ammonia makes it effective in producing an environment in
which it can survive. This leads to precipitation of organic and inorganic
compounds, which leads to struvite stone formation.
• Struvite (Mineral produced by bacteria) stone formation can be sustained only
when ammonia production is increased and the urine pH is elevated to decrease
the solubility of phosphate.
• Urease metabolizes urea into ammonia and carbon dioxide.
Urea → 2NH3 + CO2

PROTEUS INFECTION:
UTI Which progresses to :-
• Urethritis-infection of urethra
• Cystitis-inflammation of bladder
• Pyelonephritis -can be considered a progression of disease & symptoms are
therefore more profound.

SYMPTOMS:
Symptoms of pyelonephritis include flank pain , nausea & vomiting ,fever , costo
vertebral angle tenderness, hematuria & pyuria are frequently encountered.

Laboratory Diagnosis & Identification:

→The sample used for the isolation and identification of the Proteus species
depends on the nature of the disease/site of infections.
→ For UTI, midstream urine sample is used. vFor pyogenic lesions, its the pus
aspirate.
→ Sample should be collected in the sterile container maintaining aseptic
conditions and should reach laboratory within an hour of collections.

CULTURE CHARACTERISTIC:
→Blood Agar
P.mirabilis and P.vulgaris => does not form distinctive colonies on blood agar
,instead the bacteria swarm across the surface of the agar.
SWARMING
Swarming is described as the formation of concentric zonesof bacterial growth ,
able to cover the whole surface of solid culture medium as a consequence of the
organisms active motility.

ANTI-SWARMING AGENTS:
• This will physically restrict the movement of proteus cells By:
• 1- Increasing the agar concentration to 3-4% w/v
• 2- Affecting the flagella through incorporation with ethanol 5.5% , Boric acid
0.1% , detergents, bile salts etc..
• Retarding the cell growth rate by incorprtaing with growth inhibitors such as
sulfonamide ,purine bases, neomycin, sodium azide or chloral hydrite.
• Activated charcoal inhibit swarming without affecting motility.

Dienes phenomenon:
→ when two identical Proteus cultures are inoculated at different points on the
same plate of non-inhibitory medium, the resulting swarming of growth coalesce
without signs of demarcation.
→ When, however, two different strains of Proteus are inoculated, the spreading
films of growth fail to coalesce and remain separated by a narrow easily visible
area.
→ The observation of this appearance, the Dienes phenomenon has been used to
determine the identity or non-identity of strains in epidemiological studies.

Macconkey Agar
Proteus does not ferment lactose so acid is not produced and indicator will not
change color to pink organisms appears as colorless colonies (non lactose
fermenter)

Biochemical Tests:
Motility Test:
• Used to determine the motility of an organism whether it is equipped with
flagella and thus capable of swimming away from a stab mark.
• For example :
• Positive for proteus spp.
• Negative for S.aureus

INDOLE TEST:
P. mirabilis can be differentiad from P. vulgaris by INDOLE Test.
Results:
P. mirabilis => Negative
P. vulgaris => Positive

OXIDASE TEST:
• The oxidase test is used to determine if a bacterium produces cytochrome c
oxidases. The reagent turns dark blue when oxidized (oxidase positive) . The
reagent is colorless when reduced when reduced( oxidase negative )
Proteus species is oxidase negative = > colorless

PPA TEST:
Phenylalanine deaminase test also known as phenyl pyruvic acid (PPA) test is
used to test the ability of an organism to produce enzyme deaminase. This
enzyme removes the amine group from the amino acid phenylalanine and
produces phenyl pyruvic acid (PPA) and ammonia i.e. oxidative deamination of
phenylalanine. Phenyl pyruvic acid reacts with ferric iron (10% ferric chloride is
added in the medium) producing a visible green colour.

TREATMENT:
PROTEUS is sensitive to these antibiotics :
• beta-lactame except penicillin G & oxacillin
• aminoglycoside , ceftriaxone , fluoroquinolones , & trimethoprim-
sulfamethoxazole that can be used as a treatment for most proteus infection.