Microbio Lab

8/6/2019 Microbio Lab

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Spherical

Diplococcus Pneumoniae

Diplococcus Pneumoniae is a minute, slightly lancet-shaped, non-motile, non-liquefying, optionally anaerobic diplococcus. Usually occurs in pairs, surrounded by acapsule that is not present when the organism is grown on culture medium.

It is found in the sputum of lobar pneumonia, in the exudate of meningitis, andsometimes in the saliva of healthy people. Is the common cause of croupouspneumonia, but is also found in inflammations of theserous membranes.

Stains

Ordinary methods and Gram's.

Fig. 83. - Gonococci in Urethral Pus (McFarland).

Culture

Grows best at 370 C, but has a range from 240 to 420 C. Will grow upon allculture media except potato. Gelatin plates (15 per cent, gelatin) give colonies thatare small, round, circumscribed white points. On agar-agar the growth is almost

invisible.
http://chestofbooks.com/health/disease/Pathology/Chapter-VII-Inflammation-And-Regeneration.htmlhttp://chestofbooks.com/health/disease/Pathology/Serous-Or-Edematous-Infiltration.htmlhttp://chestofbooks.com/health/disease/Pathology/Serous-Or-Edematous-Infiltration.htmlhttp://chestofbooks.com/health/disease/Pathology/Chapter-VII-Inflammation-And-Regeneration.html


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Streptococcus

The RightHealth Community

Author: Steven Miller

Streptococcus is a spherical positive bacterium that is responsible for causingmeningitis, bacterial pneumonia, endocarditis, erysipelas and flesh eating bacteriacalled necrotizing fasciitis. There are some non-pathogenic streptococcus strand andthey are present in some varieties of cheese and the human mouth, skin, intestines,and upper respiratory tract.

When streptococcus bacteria are stained by the Gram Staining it turns dark

blue or violet. This means that this bacterium doesn't have an outer membrane likeother types of bacteria. This particular bacteria is also known to cause hemolysis andthis means that this particular bacteria can break down blood cells by digesting them.There are non-hemolytic streptococci and they rarely cause illness.

Streptococcus also is broken down in various groups which include A, B, C, Dand G. These groups define different kinds of the bacteria strain and the problemsthat they pose to the health of humans. Many strains of streptococcus can be treatedwith antibiotics.


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Staphylococcus

Staphylococcus, bunch of grapes", is a genus ofGram-positivebacteria.Under the microscope they appear round (cocci), and form in grape-like clusters.The Staphylococcusgenus includes at least forty species. Of these, nine have twosubspecies and one has three subspecies. Most are harmless and reside normallyon the skin and mucous membranes of humans and other organisms. Foundworldwide, they are a small component of soil microbial flora.

Spherical Gram-positive parasitic bacteria that tend to form irregular colonies;

some cause boils, septicemia or infections.

Sarcina lutea

Sarcina lutea is an older name (not used anymore) for Micrococcus luteus. It

is a Gram positive bacterium in the Firmicutes phylum. It is found in soil and air, andcan also live on human skin, and in the mouth. Micrococcus luteus is a Gram-positive, spherical, saprotrophicbacterium that belongs to the
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family Micrococcaceae. An obligate aerobe, M. luteus is found in soil, dust, waterand air, and as part of the normal flora of the mammalian skin. The bacterium alsocolonizes the human mouth, mucosae,oropharynx and upper respiratory tract.

Considered a contaminant in sick patients, M. luteus is resistant to

reduced water potential and can tolerate drying and high salt concentrations.M. luteus is coagulase negative, bacitracin susceptible, and forms bright

yellow colonies on nutrient agar. To confirm it is not Staphylococcus aureus, abacitracin susceptibility test can be performed.

M. luteus has been shown to survive in oligotrophic environments forextended periods of time. Recent work by Greenblat et al. demonstratethat Micrococcus luteus has survived for at least 34,000 to 170,000 years on thebasis of 16S rRNA analysis, and possibly much longer. [Micrococcus luteus wasformerly known as Micrococcus lysodeikticus.Micrococcus luteus reclassifyas Kocuria rhizophila.

Rod-shaped

Bacillus subtilis

Bacillus subtilis, known also as the hay bacillus orgrass bacillus, isa Gram-positive, catalase-positive bacterium commonly found in soil. A member ofthe genusBacillus, B. subtilis is rod-shaped, and has the ability to form a tough,protective endospore, allowing the organism to tolerate extreme environmentalconditions. Unlike several other well-known species, B. subtilis has historically beenclassified as an obligate aerobe, though recent research has demonstrated that thisis not strictly correct.
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Clostridium botulinum

Clostridium botulinum is a Gram-positive, rod-shaped bacterium thatproduces neurotoxins, known as botulinum neurotoxins types A-G, that cause theflaccid muscularparalysis seen in botulism. It is also the main paralytic agentin botox. C. botulinum is an anaerobic spore-former, which produces oval,subterminal endospores and is commonly found in soil.

Clostridium botulinum is a rod-shaped microorganism. It is an obligateanaerobe, meaning that oxygen is poisonous to the cells. However, C.botulinumtolerates traces of oxygen due to the enzyme superoxide dismutase (SOD)which is an important antioxidant defense in nearly all cells exposed to oxygen.C.botulinum is only able to produce the neurotoxin during sporulation, which can onlyhappen in an anaerobic environment. Other bacterial species produce spores in anunfavorable growth environment to preserve the organism's viability and permit

survival in a dormant state until the spores are exposed to favorable conditions.

In the laboratory Clostridium botulinum is usually isolated in tryptosesulfite cycloserine (TSC) growth media in an anaerobic environment with less than2% of oxygen. This can be achieved by several commercial kits that use a chemicalreaction to replace O2 with CO2 (E.J. GasPak System). C. botulinumisa lipase negative microorganism that grows between pH of 4.8 and 7 and it can'tuse lactose as a primary carbon source, characteristics important during abiochemical identification.
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Corynebacterium diphtheria

Corynebacterium diphtheriae is a pathogenic bacterium thatcauses diphtheria. It is also known as the Klebs-Lffler bacillus, because it wasdiscovered in 1884 by GermanbacteriologistsEdwin Klebs (1834 1912)and Friedrich Lffler(1852 1915).

Escherichia coli
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Escherichia coli is a Gram-negative, rod-shapedbacterium that is commonlyfound in the lowerintestine ofwarm-blooded organisms (endotherms). Most E.colistrains are harmless, but some serotypes can cause serious foodpoisoning in humans, and are occasionally responsible forproduct recalls. Theharmless strains are part of the normal flora of the gut, and can benefit their hosts by

producing vitamin K2, and by preventing the establishment ofpathogenicbacteriawithin the intestine.[4][5]

E. coliand related bacteria constitute about 0.1% ofgut flora, and fecal-oraltransmission is the major route through which pathogenic strains of the bacteriumcause disease. Cells are able to survive outside the body for a limited amount oftime, which makes them ideal indicator organisms to test environmental samplesforfecal contamination. The bacterium can also be grown easily and inexpensively ina laboratory setting, and has been intensively investigated for over 60 years. E.coliis the most widely studied prokaryotic model organism, and an important speciesin the fields ofbiotechnology and microbiology, where it has served as the hostorganism for the majority of work with recombinant DNA.

German peadiatrician and bacteriologist Theodor Escherich discovered E. coliin1885, and it is now classified as part of the Enterobacteriaceae family ofgamma-proteobacteria.

Mycobacterium tuberculosis

Mycobacterium tuberculosis (MTB) is a pathogenicbacterial species in thegenus Mycobacterium and the causative agent of most cases oftuberculosis.First
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discovered in 1882 by Robert Koch, M. tuberculosis has an unusual, waxy coatingon the cell surface (primarily mycolic acid), which makes the cells imperviousto Gram staining so acid-fast detection techniques are used instead. The physiologyofM. tuberculosis is highlyaerobic and requires high levels of oxygen. Primarily apathogen of the mammalian respiratory system, MTB infects the lungs. The most

frequently used diagnostic methods for TB are the tuberculin skin test, acid-faststain, and chest radiographs.

The M. tuberculosisgenome was sequenced in 1998.

Proteus vulgaris

Proteus vulgaris is a rod-shaped, Gram negativebacterium that inhabits theintestinal tracts of humans and animals. It can be found in soil, water and fecalmatter. It is grouped with the enterobacteriaceae and is an opportunistic pathogen ofhumans. It is known to cause urinary tract infections and wound infections.

The term Proteus signifies changeability of form, as personified in theHomeric poems in Proteus, "the old man of the sea," who tends the sealflocks ofPoseidon and has the gift of endless transformation. The first use of the termProteus in bacteriological nomenclature was made by Hauser (1885) whodescribed under this term three types of organisms which he isolated from putrefiedmeat. One of the three species Hauser identified was Proteus vulgaris so thisorganism has a long history in Microbiology.

Over the past two decades the genus Proteus, and in particularP. vulgaris,has undergone a number of major taxonomic revisions. In 1982, P. vulgaris wasseparated into three biogroups on the basis ofindole production. Biogroup one was

indole negative and represented a new species: P. penneri; while biogroup two andthree remained together as P. vulgaris.
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Spirals

Spiral shaped cells can be one of two types: either rigid called spirilla (singularspirillum) or flexible called spirochaetes (singular spirochaete). Spiral-shapedbacteria are distinguished by their length, the number and size of the spirals, anddirection of the coil. Short segments or incomplete spirals are common, as thecomma-shaped Vibrios. The spirochetes ofsyphilisare typical spiral bacteria.Diseases caused by spirochaetes include the following: syphilis, yaws, leptosporosis,and Lyme disease.

Spirals come in one of three forms, a vibrio, a spirillum, or a spirochete.

A. vibrio: a curved or comma-shaped rod

Vibrio is a genus ofGram-negativebacteria possessing a curved rodshape, several species of which can cause food borne, usually associated with

eating undercooked seafood. Typically found in saltwater, Vibrio is facultative

anaerobes that test positive foroxidase and do not form spores. All members of the

genus are motile and have polarflagella with sheaths. Recent phylogenies have

been constructed based on a suite of genes (multi-locus sequence analysis).

The name Vibrio derives from Filippo Pacini who isolated microorganisms he

called "vibrions" from cholera patients in 1854.
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Pathogenic strains

Several species ofVibrio are pathogens. Most disease causing strains are

associated with gastroenteritis but can also infect open wounds and

cause septicemia. It can be carried by numerous sea-living animals, such as crabsor prawns, and has been known to cause fatal infections in humans during exposure.

Pathogenic Vibrio include V. cholerae (the causative agent ofcholera), V.

parahaemolyticus, and V. vulnificus.Vibrio cholerae is generally transmitted via

contaminated water. Pathogenic Vibrio can cause foodborne infection, usually

associated with eating undercooked seafood.

Vibrio vulnificus outbreaks commonly occur in warm climates and small,

generally lethal, outbreaks occur regularly. An outbreak occurred in New Orleans

after Hurricane Katrina and several lethal cases occur most years in Florida.

V. parahaemolyticus is also associated with the Kanagawa phenomenon, in

which strains isolated from humanhosts (clinical isolates) are hemolytic on blood

agar plates, while those isolated from non-human sources are non-hemolytic.

Many Vibrio are also zoonotic. They cause disease in fish and shellfish, and

are common causes of mortality among domestic marine life.

Other strains

Vibrio fischeri, Photobacterium phosphoreum, and V. harveyiare notable for

their ability to communicate. BothV. fischeriandPh. phosphoreum are symbiotes of

other marine organisms (typicallyjellyfish, fish, orsquid), and produce light

via bioluminescence through the mechanism ofquorum sensing.Vibrio harveyiis a

pathogen of several aquatic animals and notable as a cause of luminous vibriosis in

shrimps (prawns)

Flagella

The "typical", early-discovered Vibrio such as V. cholerae have a single polar

flagellum (monotrichous) with sheath. Some species such as V.

parahaemolyticus and V. alginolyticus have both a single polar flagellum with sheath

and thin flagella projecting in all directions (peritrichous), and the other species such

as V. fischerihave tufts of polar flagella with sheath (lophotrichous).

B. spirillum: a thick, rigid spiral
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Spirillum in microbiology refers to a bacterium with a cell body that twists like

a spiral. It is the third distinct bacterial cell shape type besides coccus and bacillus

cells. Spirillum is a genus ofGram-negative bacteria.

Spirillum minus is associated with rat-bite fever.

Appearance

It is a genus comprising elongated forms having tufts offlagellae at both poles

and usually living in stagnant water rich in organic matter. They are twisted and

aerobic; certain species are pathogenic for humans.

It is the type genus for the family Spirilliaceae in some bacterial

classifications.

C. spirochete: a thin, flexible spiral
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Spirochaetes (also spelled spirochetes) belong to a phylum of

distinctive Gram-negativebacteria, which have long, helically coiled (spiral-shaped)

cells. Spirochaetes are chemoheterotrophic in nature, with lengths between 5 and

250 m and diameters around 0.1-0.6 m.

Spirochaetes are distinguished from other bacterial phyla by the location of

theirflagella, sometimes called axial filaments, which run lengthwise between the

bacterial inner membrane and outer membrane in periplasmic space. These cause a

twisting motion which allows the spirochaete to move about. When reproducing, a

spirochaete will undergo asexual transverse binary fission.

Most spirochaetes are free-living and anaerobic, but there are numerous exceptions.

Classification

The spirochaetes are divided into three families

(Brachyspiraceae, Leptospiraceae, and Spirochaetaceae), all placed within a single

order (Spirochaetales). Disease-causing members of this phylum include the

following:

Leptospira species, which causes leptospirosis

Borrelia burgdorferi, which causes Lyme disease

Borrelia recurrentis, which causes relapsing fever

Treponema pallidum subspeciespallidum, which causes syphilis

Treponema pallidum subspeciespertenue, which causes yaws

Brachyspira pilosicoliand Brachyspira aalborgi, which cause intestinal

spirochetosis
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Cavalier-Smith has postulated that the Spirochaetes belong in a

largerclade called Gracilicutes.

Historical

Salvarsan, the first partially organic antimicrobial drug in medical history, was

effective against spirochaetes only and was primarily used to cure syphilis.

Leptospira

Leptospira (Greekleptos, "fine, thin" and Latinspira, "coil") is a genus

ofspirochaete bacteria, including a small number

ofpathogenic and saprophytic species. Leptospira was first observed in 1907

in kidney tissue slices of a leptospirosis victim who was described as having died of

"yellow fever."

Taxonomy
http://en.wikipedia.org/wiki/Cavalier-Smithhttp://en.wikipedia.org/wiki/Cladehttp://en.wikipedia.org/wiki/Gracilicuteshttp://en.wikipedia.org/wiki/Salvarsanhttp://en.wikipedia.org/wiki/Antimicrobial_drughttp://en.wikipedia.org/wiki/Syphilishttp://en.wikipedia.org/wiki/Greek_languagehttp://en.wikipedia.org/wiki/Latin_languagehttp://en.wikipedia.org/wiki/Spirochaetehttp://en.wikipedia.org/wiki/Pathogenichttp://en.wikipedia.org/wiki/Saprophytichttp://en.wikipedia.org/wiki/Kidneyhttp://en.wikipedia.org/wiki/Leptospirosishttp://en.wikipedia.org/wiki/Yellow_feverhttp://en.wikipedia.org/wiki/Cavalier-Smithhttp://en.wikipedia.org/wiki/Cladehttp://en.wikipedia.org/wiki/Gracilicuteshttp://en.wikipedia.org/wiki/Salvarsanhttp://en.wikipedia.org/wiki/Antimicrobial_drughttp://en.wikipedia.org/wiki/Syphilishttp://en.wikipedia.org/wiki/Greek_languagehttp://en.wikipedia.org/wiki/Latin_languagehttp://en.wikipedia.org/wiki/Spirochaetehttp://en.wikipedia.org/wiki/Pathogenichttp://en.wikipedia.org/wiki/Saprophytichttp://en.wikipedia.org/wiki/Kidneyhttp://en.wikipedia.org/wiki/Leptospirosishttp://en.wikipedia.org/wiki/Yellow_fever


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Genus species serovar Serovar_name

For example:

Leptospira interrogans serovar Australis

Leptospira biflexa serovar Patoc

Morphology

Although over 200 serotypes ofLeptospira have been described, all members

of the genus have similar morphology. Leptospira are spiral-shaped bacteria that are

6-20 m long and 0.1 m in diameter with a wavelength of about 0.5 m. One or

both ends of the spirochete are usually hooked. Because they are so thin,

live Leptospira are best observed by darkfield microscopy.

The bacteria have a number of degrees of freedom; when ready to proliferate

via binary fission, the bacterium noticeably bends in the place of the future split.

Cellular structure

Leptospira have a Gram-negative-like cell envelope consisting of a

cytoplasmic and outer membrane. However, the peptidoglycan layer is associated

with the cytoplasmic rather than the outer membrane, an arrangement that is uniqueto spirochetes. The two flagella ofLeptospiraextend from the cytoplasmic membrane

at the ends of the bacteria into theperiplasmic space are necessary for the motility

ofLeptospira.

The outer membrane contains a variety of lipoproteins and

transmembraneouter membrane proteins. As expected, the protein composition of

the outer membrane differs when comparing Leptospira growing in artificial medium

with Leptospira present in an infected animal. Several leptospiral outer membrane

proteins have been shown to attach to the hostextracellular matrix and to factor H.

These proteins may be important foradhesion ofLeptospira to host tissues and in

resisting complement, respectively.

The outer membrane ofLeptospira, like those of most other Gram-negative

bacteria, contains lipopolysaccharide (LPS). Differences in the highly immunogenic

LPS structure account for the numerous serovars ofLeptospira. Consequently,

immunity is serovar specific; current leptospiral vaccines, which consist of one or

several serovars ofLeptospiraendemic in the population to be immunized, protect

only against the serovars contained in the vaccine preparation. Leptospiral LPS has

low endotoxin activity. An unusual feature of leptospiral LPS is that it activates host
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binding protein, LipL41, may account for their ability to use hemin as a source of

iron. Although they do not secrete siderophores, L. biflexa and L. interrogans may be

capable of obtaining iron from siderophores secreted by other microorganisms.

Genome

The genome of pathogenic Leptospira consists of two chromosomes. The size

of the genomes ofL. interrogans serovars Copenhageni and Lai is approximately 4.6

Mb. However, the genome ofL. borgpeterseniiserovar Hardjo is only 3.9 Mb in size

with a large number of pseudogenes, gene fragments, and insertion

sequences relative to the genomes ofL. interrogans.L. interrogans and L.

borgpeterseniishare 2708 genes from which 656 are pathogenic specific genes. The

guanine plus cytosine (GC) content is between 35% and 41%. L.

borgpeterseniiserovar Hardjo is usually transmitted by direct exposure to infected

tissues, whereas L. interrogans is often acquired from water or soil contaminated by

the urine of carrier animals harboring Leptospirain their kidneys. The high number of

defective genes and insertion sequences in L. borgpeterseniiHardjo together with

the poor survival outside of the host and difference in transmission patterns

compared to L. interrogans suggest that L. borgpeterseniiis undergoing insertion-

sequence mediated genomic decay, with ongoing loss of genes necessary for

survival outside of the host animal.

Genotyping

Genome sequence determination (of Leptospira) lead to the development

ofMultilocus sequence typing (MLST) based scheme for species level identification

of pathogenic Leptospira species. The pioneering MLST method developed by Niyaz

Ahmed in Hyderabad, India and colleagues is widely used for molecular

epidemiology studies and holds the potential to replace the highly

ambiguous, serotyping method currently in vogue for leptospiral strain identification.

Borrelia
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Borrelia is a genus ofbacteria of the spirochete phylum. It causes borreliosis,

a zoonotic, vector-borne diseasetransmitted primarily by ticks and some by lice,

depending on the species. There are 36 known species ofBorrelia.

Lyme disease

Of the 36 known species ofBorrelia, 12 of these species are known to

cause Lyme disease orborreliosis and are transmitted by ticks. The

majorBorrelia species causing Lyme disease are Borrelia burgdorferi, Borrelia

afzelii, Borrelia gariniiand Borrelia valaisiana.

Relapsing fever

Relapsing fever borreliosis often occurs with severe bacteremia. Borrelia

recurrentis is transmitted by the human body louse; no other animal reservoir ofB.

recurrentis is known. Lice that feed on infected humans acquire

the Borreliaorganisms that then multiply in the gut of the louse. When an infected

louse feeds on an uninfected human, the organism gains access when the victim

crushes the louse or scratches the area where the louse is feeding. B.

recurrentis infects the person via mucous membranes and then invades the

bloodstream.

Other tick-borne relapsing infections are acquired from other species, such

as Borrelia hermsiiorBorrelia parkeri, which can be spread from rodents, and serve
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as a reservoir for the infection, via a tick vector. Borrelia hermsiiand Borrelia

recurrentis cause very similar diseases, although the disease associated

withBorrelia hermsiihas more relapses and is responsible for more fatalities, while

the disease caused by B. recurrentis has longer febrile and afebrile intervals and a

longer incubation period.

Laboratory test

Immunoflourascent or confirm by serology by observing the organism in blood

of patient.

Genetics

All members of the Borrelia genus that have been examined harbor a

linearchromosome that is about 900 kbp in length as well as a plethora of both linear

and circularplasmids in the 5-220 kbp size range. Genome sequences have been

determined forB. burgdorferi, B. garinii, B. afzelii, B. duttoniiand B. recurrentis. The

chromosomes, which carry the vast majority of the housekeeping genes, appear to

be very constant in gene content and organization across the genus. The content of

the plasmids, which carry most of the genes that encode the differentially-expressed

surface proteins that interact with Borrelia'sarthropod andvertebrate hosts, are much

more variable. B. burgdorferistrain B31, the B. burgdorferitype strain, has been

studied in the most detail and harbors twelve linear and nine circular plasmids thatcomprise about 612 kbp. The plasmids are unusual, as compared to most bacterial

plasmids, in that they contain many paralogous sequences, a large number of

pseudogenes and, in some cases, essential genes. In addition, a number of the

plasmids have features suggesting that they are prophages. Some correlations

between genome content and pathogenicity have been deduced and comparative

whole genome analyses promise future progress in this arena.
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