Hemolysis is the breakdown of

red blood cell Red blood cells (RBCs), referred to as erythrocytes (, with -''cyte'' translated as 'cell' in modern usage) in academia and medical publishing, also known as red cells, erythroid cells, and rarely haematids, are the most common type of blood cel ...

s. The ability of bacterial colonies to induce hemolysis when grown on

blood agar An agar plate is a Petri dish that contains a growth medium solidified with agar, used to Microbiological culture, culture microorganisms. Sometimes selective compounds are added to influence growth, such as antibiotics. Individual microorganism ...

is used to classify certain

microorganism A microorganism, or microbe, is an organism of microscopic scale, microscopic size, which may exist in its unicellular organism, single-celled form or as a Colony (biology)#Microbial colonies, colony of cells. The possible existence of unseen ...

s. This is particularly useful in classifying

streptococcal ''Streptococcus'' is a genus of gram-positive spherical bacteria that belongs to the family Streptococcaceae, within the order Lactobacillales (lactic acid bacteria), in the phylum Bacillota. Cell division in streptococci occurs along a single ...

species. A substance that causes hemolysis is called a

hemolysin Hemolysins or haemolysins are lipids and proteins that cause lysis of red blood cells by disrupting the cell membrane. Although the lytic activity of some microbe-derived hemolysins on red blood cells may be of great importance for nutrient acqui ...

Types

Alpha-hemolysis

When alpha-hemolysis (α-hemolysis) is present, the agar under the colony is light and greenish. ''

Streptococcus pneumoniae ''Streptococcus pneumoniae'', or pneumococcus, is a Gram-positive, spherical bacteria, hemolysis (microbiology), alpha-hemolytic member of the genus ''Streptococcus''. ''S. pneumoniae'' cells are usually found in pairs (diplococci) and do not f ...

'' and a group of oral streptococci ( ''Streptococcus'' viridans or viridans streptococci) display alpha-hemolysis. This is sometimes called ''green hemolysis'' because of the color change in the agar. Other synonymous terms are ''incomplete hemolysis'' and ''partial hemolysis''. Alpha-hemolysis is caused by the bacteria's production of

hydrogen peroxide Hydrogen peroxide is a chemical compound with the formula . In its pure form, it is a very pale blue liquid that is slightly more viscosity, viscous than Properties of water, water. It is used as an oxidizer, bleaching agent, and antiseptic, usua ...

, which oxidizes

hemoglobin Hemoglobin (haemoglobin, Hb or Hgb) is a protein containing iron that facilitates the transportation of oxygen in red blood cells. Almost all vertebrates contain hemoglobin, with the sole exception of the fish family Channichthyidae. Hemoglobin ...

and produces the green oxidized derivative

methemoglobin Methemoglobin (British: methaemoglobin, shortened MetHb) (pronounced "met-hemoglobin") is a hemoglobin ''in the form of metalloprotein'', in which the iron in the heme group is in the Fe3+ (ferric) state, not the Fe2+ (ferrous) of normal hemoglobin ...

Beta-hemolysis

Beta-hemolysis (β-hemolysis), sometimes called ''complete hemolysis'', is a complete

lysis Lysis ( ; from Greek 'loosening') is the breaking down of the membrane of a cell, often by viral, enzymic, or osmotic (that is, "lytic" ) mechanisms that compromise its integrity. A fluid containing the contents of lysed cells is called a ...

of red cells in the media around and under the colonies: the area appears lightened (yellow) and transparent.

Streptolysin Streptolysins are two homogenous exotoxins from ''Streptococcus pyogenes''. Types include streptolysin O (SLO; ''slo''), which is oxygen-labile, and streptolysin S (SLS; ''sagA''), which is oxygen-stable. SLO is part of the thiol-activated c ...

, an

exotoxin An exotoxin is a toxin secreted by bacteria. An exotoxin can cause damage to the host by destroying cells or disrupting normal cellular metabolism. They are highly potent and can cause major damage to the host. Exotoxins may be secreted, or, sim ...

, is the enzyme produced by the bacteria which causes the complete lysis of red blood cells. There are two types of streptolysin: streptolysin O (SLO) and streptolysin S (SLS). Streptolysin O is an oxygen-sensitive cytotoxin secreted by most Group A streptococcus (GAS) and ''

Streptococcus dysgalactiae ''Streptococcus dysgalactiae'' is a gram positive, beta-haemolytic, coccal bacterium belonging to the family ''Streptococcaceae''. It is capable of infecting both humans and animals, but is most frequently encountered as a commensal of the alim ...

''; it interacts with cholesterol in the membrane of eukaryotic cells (mainly red and white blood cells, macrophages, and platelets), usually resulting in β-hemolysis under the surface of blood agar. Streptolysin S is an oxygen-stable cytotoxin also produced by most GAS strains which results in clearing on the surface of blood agar. SLS affects immune cells, including

polymorphonuclear leukocyte Granulocytes are cells in the innate immune system characterized by the presence of specific granules in their cytoplasm. Such granules distinguish them from the various agranulocytes. All myeloblastic granulocytes are polymorphonuclear, that is, ...

s and lymphocytes, and is thought to prevent the host immune system from clearing infection. ''

Streptococcus pyogenes ''Streptococcus pyogenes'' is a species of Gram-positive, aerotolerant bacteria in the genus '' Streptococcus''. These bacteria are extracellular, and made up of non-motile and non-sporing cocci (round cells) that tend to link in chains. They ...

'', or Group A beta-hemolytic Strep (GAS), as well as ''

Streptococcus agalactiae ''Streptococcus agalactiae'' (also known as group B streptococcus or GBS) is a gram-positive coccus (round bacterium) with a tendency to form chains (as reflected by the genus name ''Streptococcus''). It is a beta-hemolytic, catalase-negative, an ...

'' display beta-hemolysis. The hemolysis of some weakly beta-hemolytic organisms is enhanced when streaked close to certain beta hemolytic strains of ''

Staphylococcus aureus ''Staphylococcus aureus'' is a Gram-positive spherically shaped bacterium, a member of the Bacillota, and is a usual member of the microbiota of the body, frequently found in the upper respiratory tract and on the skin. It is often posi ...

''. This phenomenon is the mechanism behind the ''

CAMP test The CAMP test (Christie–Atkins–Munch-Petersen) is a test to identify group B β-hemolytic streptococci ('' Streptococcus agalactiae'') based on their formation of a substance, CAMP factor, that enlarges the area of hemolysis formed by the � ...

'', a test that was historically used for the identification of ''

'' and ''

Listeria monocytogenes ''Listeria monocytogenes'' is the species of pathogenic bacteria that causes the infection listeriosis. It is a facultative anaerobic bacterium, capable of surviving in the presence or absence of oxygen. It can grow and reproduce inside the ho ...

''. A modified version of this test called the reverse CAMP test, utilizing ''S. agalactiae'' instead of ''S. aureus'', can also be used to identify ''

Clostridium perfringens ''Clostridium perfringens'' (formerly known as ''C. welchii'', or ''Bacillus welchii'') is a Gram-positive, bacillus (rod-shaped), anaerobic, spore-forming pathogenic bacterium of the genus '' Clostridium''. ''C. perfringens'' is ever-present ...

''.

Gamma-hemolysis

If an organism does not induce hemolysis, the agar under and around the colony is unchanged and the organism is called ''non-hemolytic'' or said to display gamma-hemolysis (γ-hemolysis). ''

Enterococcus faecalis ''Enterococcus faecalis'' – formerly classified as part of the group D '' Streptococcus,'' is a Gram-positive, commensal bacterium naturally inhabiting the gastrointestinal tracts of humans. Like other species in the genus '' Enterococcus'', ' ...

'' (formerly called "Group D Strep"), ''

Staphylococcus saprophyticus ''Staphylococcus saprophyticus'' is a Gram-positive coccus belonging to the genus ''Staphylococcus''. ''S. saprophyticus'' is a common cause of community-acquired urinary tract infections. History ''Staphylococcus saprophyticus'' was not recogni ...

'', and ''

Staphylococcus epidermidis ''Staphylococcus epidermidis'' is a Gram-positive bacterium, and one of over 40 species belonging to the genus ''Staphylococcus''. It is part of the human flora, normal human microbiota, typically the skin flora, skin microbiota, and less commonl ...

'' display gamma-hemolysis.

Hemodigestion

When some otherwise non-hemolytic bacteria, such as strains of the cholera-causing bacteria ''

Vibrio cholerae ''Vibrio cholerae'' is a species of Gram-negative bacteria, Gram-negative, Facultative anaerobic organism, facultative anaerobe and Vibrio, comma-shaped bacteria. The bacteria naturally live in Brackish water, brackish or saltwater where they att ...

'', are plated on blood agar, no clearings are observed surrounding the isolated colonies, but the blood surrounding larger areas of growth turns green. This process, called hemodigestion, is caused by the metabolic by-products of the bacteria in aerobic conditions.

Notes

References

* *Kato, Gregory J.; Steinberg, Martin H.; Gladwin, Mark T. (2017-03-01). "Intravascular hemolysis and the pathophysiology of sickle cell disease". ''Journal of Clinical Investigation''. 127 (3): 750–760. doi:10.1172/JCI89741.

ISSN An International Standard Serial Number (ISSN) is an eight-digit to uniquely identify a periodical publication (periodical), such as a magazine. The ISSN is especially helpful in distinguishing between serials with the same title. ISSNs a ...

0021-9738. {{Clinical microbiology techniques Blood Microbiology terms