Swarming Growth of Proteus on Blood Agar: Description of Swarming and Proteus Details
Swarming Growth of Proteus on Blood Agar
Swarming growth of Proteus on blood agar as shown above image. The morphological events associated with the swarming phenomenon of Proteus have been described by a number of investigators. From these studies, it is possible to describe the events that normally occur during the process. Broth cultures of Proteus normally consist of very short rods about 0.6µm wide and 1-2 µm long. Filamentous forms in broth have been reported, but their presence seems to be associated with variant types to be discussed later. When a suitable solid medium is centrally inoculated with a suspension of the very short rods, the cells enlarge slightly, forming rods about 0.8 µm wide and 2-4 µm long. These cells undergo normal division and growth, producing a typical bacterial colony. After a period of time that depends on culture conditions, some of the cells near the perimeter of the colony begin to undergo a dramatic morphological change to highly elongated forms that are about 0.7µm wide and 20-80 µm long. This increase in cell length is associated with a corresponding striking increase in flagellation. Hoeniger has characterized the development of flagella associated with the process of swarming. The short cells produced by the division of the swarm cells continue to grow and divide, and the cycle is repeated with a second swarm band emerging from the edge of the first band. This process, known as the zonation phenomenon, continues until the entire surface of the plate is covered by several concentric bands of heavy and light growth as shown above picture.
Proteus General Characteristic
They are Gram-negative rods and facultative anaerobes.
They are motile due to peritrichous flagella, non encapsulated, non-spore-forming.
They are most commonly found in the human intestinal tract as part of normal human intestinal flora as well as saprophytic and are widely distributed in nature.
They are non-lactose fermenting (NLF).
The main species of medical importance are P. mirabilis and P. vulgaris. They are opportunist pathogens and may cause many types of infection.
They are actively motile, non-capsulated, Gram-negative bacilli measuring 1-3µm x 0.5 µm in size. Motility is not as easily observed at 35-37 °C at room temperature ( 20-28°C). Culture on blood agar shows a characteristics’ fishy or seminal odor’ with swarming growth as shown above picture.
MacConkey agar: Proteus produces individual non-lactose fermenting colonies after overnight incubation at 35-37°C and swarming is prevented due to bile salt present in the medium.
CLED agar: Blue-gray translucent colonies and absence of swarming are due to electrolyte deficiency.
XLD agar: Proteus produces individual non-lactose fermenting colonies after overnight incubation at 35-37°C and swarming is prevented due to bile salt present in the medium
The Proteus possess thermostable, somatic (O), and thermolabile flagellar (H) antigens upon which, several serotypes have been recognized. WeIl and Felix ( 1966) observed flagellated strains of Proteus grew on agar as a thin surface film (named Hauch) whereas non flagellated variant strains without surface film ( Ohne Hauch). They also noted that certain non-motile strains of Proteus vulgaris, called X strains were agglutinated by sera of typhus patients. The sharing of these polysaccharide antigens of Proteus witH some rickettsiae forms the basis of the Weil-Felix reaction for the diagnosis of some rickettsial infections. Total three non-motile strains, among them two of Proteus vulgaris OX2, OX19, and one Proteus mirabilis OX K are applicable as the antigens for this test.
They can cause –
Abdominal and wound infections
Infection of ear
Specimens: It depends on the site of infection and common samples are urine, pus, and ear discharge. Midstream urine (MSU) in UTI whereas pus in pyogenic lesions.
Specimens collection: The sample should be collected in a sterile container under all aseptic conditions and transported to the laboratory immediately.
Gram stain: Gram-negative rods which are non-capsulated and non-sporing ( evidence).
Culture:Swarming effect over blood agar plate as a consequence of the organisms active motility and NLF colonies on Macconkey agar whereas blue-gray translucent colonies on CLED agar with fishy or seminal odor.
Dienes phenomenon: When two different strains of Proteus species inoculate at different places of the same non-inhibitory medium (blood agar), swarming of the two strains remains separated by a narrow visible furrow. However, in the case of two identical strains of Proteus, swarming of two coalesce without signs of demarcation. Such a condition is called the Dienes phenomenon.
Hanging drop preparation: Actively motile rods observation
Methyl red (MR) Test:The methyl red test is used to identify bacteria to produce pyruvic acid from glucose metabolism. Both Proteus vulgaris and mirabils are methyl red test positive.
Voges–Proskauer (VP) Test:It is a test used to detect acetoin in bacterial broth culture. A red-brown color indicates a positive result, while a yellow-brown color indicates a negative result. Both P. vulgaris and P. mirabilis are VP negative.
Triple Sugar Iron (TSI) Test: This test is used to determine the ability of bacteria to ferment sugars and to produce hydrogen sulfide ( H2S) and gas formation.
Proteus mirabilis: Red slope/yellow butt with H2S production i.e. black and presence of gas formation
Proteus vulagris: Red slope/yellow butt with H2S production i.e. black and gas formation -different strains give different results
SIM Test:This single panel test is useful for detecting H2S production, indole detection, and motility test.
Proteus mirabilis: H2S production, indole negative and motile
Proteus vulagris: H2S production, indole positive and motile
IMViC Test: The indole test is used to determine the ability of bacteria to convert tryptophan into indole. P. mirabilis can be differentiated from P. vulgaris by the indole test in which P. mirabilis is negative whereas P. vulgaris positive.
API test: By inoculating microorganisms to a strip. After inoculation metabolic cause color change, to complete the identification of the strains either identified to the genus only or that have multiple genera consist of profile number.
Vitek system: It is a new automatic system for identification and susceptibility testing for most clinically important bacteria. It compromises: -A filter/sealer: enable -inoculation of the card within a few minutes. An inoculator /leader – A computer and printer.
Antibiotic susceptibility test (AST): AST is important as Proteus bacilli are resistant to many of the common antibiotics.
P. mirabilis has fascinated scientists for over 125 years for its ability to differentiate from short swimmer cells into elongated swarm cells that express hundreds to thousands of flagella.
Swarming is distinct from swimming motility in that it refers to multicellular flagellum-mediated migration across a surface rather than movement in a liquid medium or through soft agar.
Multiple drug-resistant (MDR) strains to carry R plasmids have become very important in nosocomial infections.
Swarming phenomenon swarming is described as the formation of concentric zones of bacterial growth, able to cover the whole surface of a solid culture medium. P. mirabilis and P. vulgaris are known for their swarming ability over blood agar as shown above image.
The distinctive characters of the genus are PPA, urease, and H2S positive.
Indole helps to differentiate P. vulgaris ( positive) from P. mirabilis ( negative).
Dienes phenomenon or typing is using successfully to determine the relationship between strains of Proteus species in studies of cross-infection.
Swarming growth is a problem In the laboratory when mixed growth is obtained in which Proteus bacilli are present with other bacteria and thus in such scenario swarming inhibition is mandatory. Several methods have been used to inhibit warming are- √Increasing agar concentration up to 6% √incorporating alcohol 5-6% √Chloral hydrate √Sodium azide √Sulphonamide √Surface active agents and √boric acid.
#Proteus swarming growth on blood agar and its identification using various biochemical tests e.g. TSI test, MIU test, Citrate Utilization test as shown below-
#Proteus growth on blood agar and McConkey agar-
#Proteus swarming growth covering other bacteria
#Swarm and vegetative cells of Proteus spp.
#Swarming growth of Proteus having swimmer and swarmer cells: Swarming growth of Proteus having swimmer and swarmer cells can be determined using Gram’s staining- i.e. swimmer cells- small-near the center of growth plate while swarmer cells- large-away the center of growth plate as shown just above the video.
# Dienes Phenomenon of typing of Proteus
#Motile bacteria under hanging drop Preparation as shown below-Testing motility of bacteria under hanging drop preparation is a recommended method. Bacteria are motile due to having flagella. The objective of this observation includes- Active motile or true motility Brownian Movement Motile bacteria and non-motile bacteria Motile bacteria are-