Name:- Zain Abbas

RollNo:- 9837
Program:- DPT(6th)
Submitted to Ma'm iqra Nawaz
Q:- Clostridium Species
Important properties
Clinical Findings
Pathogenesis
Lab diagnosis

Clostridium species
:- Clostridium species are anaerobic, fermentative, spore-
forming Gram-positive bacteria belonging to the phylum
Firmicutes. Toxin-producing species can cause mild-to-fatal
food poisoning, most famously Clostridium botulinum and
Clostridium perfringens. Several other Clostridium species can
cause meat spoilage.
:- Clostridium species (Clostridium perfringens, C. butyricum, C.
clostridioforme, C. innocuum, C. ramosum, C. septicum, C.
sordellii, C. tertium)

History:-
:- In the late 1700s, Germany experienced a number of
outbreaks of an illness that seemed connected to eating certain
sausages. In 1817, the German neurologist Justinus Kerner
detected rod-shaped cells in his investigations into this so-
called sausage poisoning. In 1897, the Belgian biology professor
Emile van Ermengem published his finding of an endospore-
forming organism he isolated from spoiled ham. Biologists
classified van Ermengem's discovery along with other known
gram-positive spore formers in the genus Bacillus. This
classification presented problems, however, because the isolate
grew only in anaerobic conditions, but Bacillus grew well in
oxygen.

:- Circa 1880, in the course of studying fermentation and

butyric acid synthesis, a scientist surnamed Prazmowski first
assigned a binomial name to Clostridium butyricum.[The
mechanisms of anaerobic respiration were still not yet well
elucidated at that time,so taxonomy of anaerobes was still
nascent.

:- In 1924, Ida A. Bengtson separated van Ermengem's

microorganisms from the Bacillus group and assigned them to
the genus Clostridium. By Bengtson's classification scheme,
Clostridium contained all of the anaerobic endospore-forming
rod-shaped bacteria, except the genus Desulfotomaculum.

:- Among more than 200 known Clostridium spp. at least 30 are

associated with human disease. They usually appear as gram-
positive rods, however many strains may appear gram-variable
or gram-negative. Loss of gram-positive appearance occurs with
direct stains of clinical material or in cultures after prolonged
incubation or in species with terminal spores. Clostridium
perfringens is the most important of the species and accounts
for 20-40% of all isolates. Speciation is based mostly on cellular
morphology, spores location (central, terminal or sub terminal),
biochemical reactions, gas liquid chromatography for
fermentation products; and the demonstration of production of
lecithinase (or alpha toxin by C. perfringens ) and lipase (by
Clostridium sporogenes, Clostridium novyi and Clostridium
botulinum.
:- Clostridium is a genus of Gram-positive bacteria. This genus
includes several significant human pathogens, including the
causative agents of botulism and tetanus. The genus formerly
included an important cause of diarrhea, Clostridioides difficile,
which was reclassified into the Clostridiodies genus in 2016.
They are obligate anaerobes capable of producing endospores.
The normal, reproducing cells of Clostridium, called the
vegetative form, are rod-shaped, which gives them their name,
from the Greek κλωστήρ or spindle. Clostridium endospores
have a distinct bowling pin or bottle shape, distinguishing them
from other bacterial endospores, which are usually ovoid in
shape. Clostridium species inhabit soils and the intestinal tract
of animals, including humans. Clostridium is a normal
inhabitant of the healthy lower reproductive tract of females.

:- The genus, as traditionally defined, contains many organisms

not closely related to its type species. The issue was originally
illustrated in full detail by a rRNA phylogeny from Collins 1994,
which split the traditional genus (now corresponding to a large
slice of Clostridia) into twenty clusters, with cluster I containing
the type species and its close relatives.Over the years, this has
resulted in many new genera being split out, with the ultimate
goal of constraining Clostridium to cluster I.

:- "Clostridium" cluster XIVa and "Clostridium" cluster IV

efficiently ferment plant polysaccharide composing dietary
fiber,making them important and abundant taxa in the rumen
and the human large intestine.As mentioned before, these
clusters are not part of current Clostridium,and use of these
terms should be avoided due to ambiguous or inconsistent
usage.

EPIDEMIOLOGY
Organisms of the genus Clostridium are important members of
the humans’ anaerobic gastrointestinal and cervical-vaginal
flora. Clostridia are ubiquitous and are found in the soil, marine
sediment, decaying vegetation and intestinal tract of humans,
other vertebrate and insects. Human infections with clostridia
can result from endogenous or exogenous infection.
:- How many species of Clostridium are there
Clostridium contains around 250 species that include common
free-living bacteria, as well as important pathogens. The main
species responsible for disease in humans are: Clostridium
botulinum can produce botulinum toxin in food or wounds and
can cause botulism.
:- Where is Clostridium found
Clostridium, genus of rod-shaped, usually gram-positive
bacteria, members of which are found in soil, water, and the
intestinal tracts of humans and other animals.
:- What disease is caused by Clostridium
Clostridium difficile (klos-TRID-e-um dif-uh-SEEL), also known as
Clostridioides difficile and often referred to as C. difficile or C.
diff, is a bacterium that can cause symptoms ranging from
diarrhea to life-threatening inflammation of the colon.

Important Properties
:- Genus of rod-shaped
:- Usually gram-positive bacteria
:- Members of which are found in soil, water, and the intestinal
tracts of humans and other animals.
:- Most species grow only in the complete absence of oxygen.
Dormant cells are highly resistant to heat, desiccation, and toxic
chemicals and detergents
:- Clostridium is a genus of gram-positive, spore-forming
bacteria belonging to the family Clostridiaceae.
:- Vegetative cells are rod shaped and arranged in pairs or
short chains. The majority of species are obligate anaerobes;
however, some species can grow under aerobic conditions or
are aerotolerant. There are close to 200 species of Clostridium,
with only a few species being pathogenic to humans. Several
species produce toxins.

Clinical Findings
:- Infections caused by these organisms range from a variety of
localized wound contamination to overwhelming systemic
disease.Clostridial histiotoxic syndromes are mediated by toxins
and include soft tissue infections such as gas gangrene (caused
by C. perfringens), enteric diseases such as clostridial food
poisoning, enteritis necroticans, antibiotic associated colitis
(caused by C. difficile ), discussed elsewhere, and neutropenic
enterocolitis (caused by C. septicum) and neurological
syndromes such as tetanus and botulism (both discussed
elsewhere).

:- C. septicum can cause spontaneous, non-traumatic gas

gangrene, and C. sordellii can induce gas gangrene of the
uterus, as a consequence of spontaneous abortion, normal
vaginal delivery and traumatic injury.Clostridial bacteremia
account for clinically significant anaerobic bacteremia second
only to Bacteroides spp. Clostridia are also often isolated from
polymicrobial intraabdominal, biliary, pleuropulmonary, central
nervous system, genitourinary and skin and soft tissue
infections.

:- Blood samples, which were obtained from patients who lived

in a rural area with approximately 500 acute-care hospital beds,
were cultured from 1990 through 1997. We retrospectively
reviewed the blood cultures that yielded Clostridium species
(74 [0.12%] of 63,296 cultures). These were obtained from 46
different hospitalized patients (incidents per hospital, 0.03%).
The source of the Clostridium species was a gastrointestinal site
in 24 patients (52.2%). The most frequently identified
Clostridium species was Clostridium perfringens (in 10 [21.7%]
of patients), followed by Clostridium septicum (in 9 [19.6%]).
Thirty-one patients (67.4%) were aged > or =65 years, 13
patients (28.3%) had diabetes mellitus, and underlying
malignancy was present in 22 patients (47.8%). The mortality
rate of patients whose condition had been managed surgically
was 33%; for those patients whose conditions required medical
management, the mortality rate was 58%. Clostridium
bacteremia in these patients usually had a gastrointestinal
source, it often occurred in patients with serious underlying
medical conditions, and it rarely was the result of traumatic
farm accidents.

Pathogenesis
:- Toxins mediate the Clostridial histiotoxic syndromes. Toxins
are biologically active proteins that are antigenic and capable of
neutralization by specific antisera. C. perfringens produces 4
major lethal toxins, based on which it is classified into 5
serological types classified A to E. Additional virulence factors
include enterotoxin, neuraminadase and haemolysins. The role
of clostridia in the pathogenesis of polmicrobial infections is
unclear.

:- Clostridium perfringens is the most common clinical isolate

of the genus. It is a ubiquitous bacterium associated with
several exotoxin-mediated clinical diseases. There are 12
recognized toxins, and the species is divided into types A
through E on the basis of the spectrum of toxins produced .
The disease syndromes caused by C. perfringens are food
poisoning, necrotizing enteritis, and gas gangrene.
Clostridium contains around 250 species that include common
free-living bacteria, as well as important pathogens.The main
species responsible for disease in humans are:-

:- Clostridium botulinum can produce botulinum toxin in food

or wounds and can cause botulism. This same toxin is known as
Botox and is used in cosmetic surgery to paralyze facial muscles
to reduce the signs of aging; it also has numerous other
therapeutic uses.

:- Clostridium perfringens causes a wide range of symptoms,

from food poisoning to cellulitis, fasciitis, necrotic enteritis and
gas gangrene.

:- Clostridium tetani causes tetanus.

:- Clostridium sordellii (now Paeniclostridium) can cause a fatal
infection in exceptionally rare cases after medical abortions.
Bacillus and Clostridium are often described as Gram-variable,
because they show an increasing number of gram-negative cells
as the culture ages.
Clostridium and Bacillus are both in the phylum Firmicutes, but
they are in different classes, orders, and families.
Microbiologists distinguish Clostridium from Bacillus by the
following features:-

:- Clostridium grows in anaerobic conditions; Bacillus grows in

aerobic conditions.

:- Clostridium forms bottle-shaped endospores; Bacillus forms

oblong endospores.

:- Clostridium does not form the enzyme catalase; Bacillus

secretes catalase to destroy toxic byproducts of oxygen
metabolism.
Clostridium and Desulfotomaculum are both in the class
Clostridia and order Clostridiales, and they both produce bottle-
shaped endospores, but they are in different families.
Clostridium can be distinguished from Desulfotomaculum on
the basis of the nutrients each genus uses (the latter requires
sulfur).
Glycolysis and fermentation of pyruvic acid by Clostridia yield
the end products butyric acid, butanol, acetone, isopropanol,
and carbon dioxide.
The Schaeffer-Fulton stain (0.5% malachite green in water) can
be used to distinguish endospores of Bacillus and Clostridium
from other microorganisms.There is a commercially available
polymerase chain reaction (PCR) test kit (Bactotype) for the
detection of C. perfringens and other pathogenic bacteria.

Lab diagnosis
:- Isolation of Clostridia from wound, pus, blood or faeces,
along with toxin and serological assays aid in the diagnosis of
clostridial infections. The significance of clostridia in
polymicrobial isolates is unknown. Clinical diagnosis is
especially important in infections such as gas gangrene in which
demonstration of clostridial myonecrosis is critical in the
diagnosis.
:- The laboratory diagnosis of Clostridium difficile-associated
disease (CDAD) is based on culture and toxin detection in fecal
specimens. Culture is performed on a commercially available
selective media. C. difficile colony morphology is typical when
viewed under a dissecting microscope. Definitive identification
is best obtained by gas liquid chromatography. Culture is very
sensitive but, when used alone without toxin testing, it leads to
low specificity and misdiagnosis of CDAD when high rates of
asymptomatic carriage exist. Toxin detection by a tissue culture
cytotoxin assay followed by neutralisation with specific
antiserum is often considered the standard. However, this
approach lacks sensitivity and has not detected up to 30% of
patients with confirmed CDAD. Multiple enzyme immunoassays
(EIAs) have been introduced by various manufacturers for the
detection of toxin A alone or for both toxins A and B. Some of
these are designed to give results in less than 1 h. Comparative
studies of EIA kits reported that the sensitivity and specificity
are slightly lower than cytotoxin assays. Toxigenic culture tests
C. difficile isolates for toxin production: colonies isolated on
selective media are tested for in-vitro toxin production either
by a cytotoxicity assay or by direct EIA. It has higher sensitivity
than the cytotoxicity assay and equivalent specificity. In the
routine laboratory, culture and toxin detection should be
performed on every specimen and, in culture-positive and fecal
toxin-negative cases, toxigenic cultures should be performed on
isolated colonies.

Uses:-
:- Clostridium thermocellum can use lignocellulosic waste and
generate ethanol, thus making it a possible candidate for use in
production of ethanol fuel. It also has no oxygen requirement
and is thermophilic, which reduces cooling cost.
:- Clostridium acetobutylicum was first used by Chaim
Weizmann to produce acetone and biobutanol from starch in
1916 for the production of cordite (smokeless gunpowder).
:- Clostridium botulinum produces a potentially lethal
neurotoxin used in a diluted form in the drug Botox, which is
carefully injected to nerves in the face, which prevents the
movement of the expressive muscles of the forehead, to delay
the wrinkling effect of aging. It is also used to treat spasmodic
torticollis and provides relief for around 12 to 16 weeks.
:- Clostridium butyricum MIYAIRI 588 strain is marketed in
Japan, Korea, and China for Clostridium difficile prophylaxis due
to its reported ability to interfere with the growth of the latter.
:- Clostridium histolyticum has been used as a source of the
enzyme collagenase, which degrades animal tissue. Clostridium
species excrete collagenase to eat through tissue and, thus,
help the pathogen spread throughout the body. The medical
profession uses collagenase for the same reason in the
débridement of infected wounds. Hyaluronidase,
deoxyribonuclease, lecithinase, leukocidin, protease, lipase,
and hemolysin are also produced by some clostridia that cause
gas gangrene.
:- Clostridium ljungdahlii, recently discovered in commercial
chicken wastes, can produce ethanol from single-carbon
sources including synthesis gas, a mixture of carbon monoxide
and hydrogen, that can be generated from the partial
combustion of either fossil fuels or biomass.
:- Clostridium butyricum converts glycerol to 1,3-propanediol.
:- Genes from Clostridium thermocellum have been inserted
into transgenic mice to allow the production of endoglucanase.
The experiment was intended to learn more about how the
digestive capacity of monogastric animals could be improved.
:- Nonpathogenic strains of Clostridium may help in the
treatment of diseases such as cancer. Research shows that
Clostridium can selectively target cancer cells. Some strains can
enter and replicate within solid tumors. Clostridium could,
therefore, be used to deliver therapeutic proteins to tumours.
This use of Clostridium has been demonstrated in a variety of
preclinical models.
:- Mixtures of Clostridium species, such as Clostridium
beijerinckii, Clostridium butyricum, and species from other
genera have been shown to produce biohydrogen from yeast
waste.

Subdivisions
Clostridium aceticum
Clostridium acetireducens
Clostridium acetobutylicum
Clostridium acidisoli
Clostridium aciditolerans
Clostridium acidurici
Clostridium aerotolerans
Clostridium aestuarii
Clostridium akagii
Clostridium aldenense
Clostridium aldrichii
Clostridium algidicarnis
Clostridium algidixylanolyticum
Clostridium algifaecis
Clostridium algoriphilum
Clostridium alkalicellulosi
Clostridium amazonense
Clostridium aminophilum
Clostridium aminovalericum
Clostridium amygdalinum
Clostridium amylolyticum
Clostridium arbusti
Clostridium arcticum
Clostridium argentinense
Clostridium asparagiforme
Clostridium aurantibutyricum
Clostridium autoethanogenum
Clostridium baratii
Clostridium bartlettii
Clostridium beijerinckii
Clostridium bifermentans
Clostridium bolteae
Clostridium bornimense
Clostridium botulinum