MIC 301/311

BACTERIOLOGY
 LECTURE 1
•Gram-Negative Rods
Enteric Bacteria:
General Properties,
Isolation, Identification,
Characteristics, Diagnostic features
and economic importance of:
 Escherichia
Salmonella
:From environment, Food and Clinical
Samples
DEFINITIONS
• What are Gram-negative bacteria?
Gram-negative bacteria (GNB) are a
large group of bacteria that do not retain
the crystal violet stain used in the Gram
staining method.
-
 Gram-Negative Rods
• The Gram negative rods are also referred
to as GN-bacilli, so described, because of
their shape. It could
-The Enterobacteriaceae
-Nonfermenting Gram-negative
Bacilli e.g Pseudomonas,
Acinetobacter, Burkholderia,
Stenotrophomonas, Achromobacter,
Shewanella etc.
 So, what Are Enteric Bacteria?
• Enteric comes from the Greek word enteron,
which literally means intestine.
• , we know they are crucial to our health. They
help digest and absorb the vitamins from our
food, and help keep us healthy. They can,
however, cause us problems as well. So, the
existence and balance of enteric bacteria is
clearly very important to our wellbeing.
 Examples of Enteric Bacteria
Enteric bacteria include:
• Faecalibacterium prausnitzii: mesophilic, anaerobic , non-spore forming and non-motile
Gram-positive rod. It is one of the most abundant and important commensal bacteria of the
human gut microbiota.

• Lactobacillus, Enterococcus, and Bacteroides can be found in the small

intestine. Escherichia coli and other coliforms can be found here as well.
• E. coli and other coliforms are more abundant in the large intestine (colon), The large
intestine also contains enterococci, lactobacilli, and clostridia. More abundantly, the
anaerobes, Bifidobacterium and Bacteroides are found here too
• Other types of bacteria that are found in the gut include :
– Staphylococcus
– Other members of the Enterobacteiaceae family
(Salmonella, Klebsiella, Enterobacter, Proteus,
– Pseudomonas
– Other anaerobes: Peptostreptococcus, and Peptococcus.

• However, the number and variety of enteric bacteria as well as their main locations can vary
depending on numerous factors, such as the person's age, diet, as well as antibiotics they
may be taking.
 The Family Enterobacteriaceae
• The Enterobacteriaceae are a large,
heterogeneous group of gram-negative rods which
are ubiquitous in nature.
• Many species can exist as free living in diverse
ecological niches, both terrestrial and aquatic
environments, and some are associated with
animals, plants, or insects only.
• Currently, the family comprises 51 genera and 238
species. Because of their abundance as members
of the gut flora, they are generally refer to as the
“Enterics”
 The Family Enterobacteriaceae
• The enterics are important because some of them
symbiotically aid the digestion of their hosts, while
other pathogenic species cause disease or death in
their host organism.
• Many are significant human, animal, and plant
pathogens causing a range of infections.

• The members include many genera such as

Escherichia, Shigella, Salmonella, Enterobacter,
Klebsiella, Serratia, Proteus etc. Escherichia coli, are
part of the normal microbiota and incidentally cause
disease (Opportunists), but others, the salmonellae
and shigellae, are regularly pathogenic for humans.
 Enterobacteriaceae cont…
• The family Enterobacteriaceae is divided into
two major groups on the basis of lactose
fermentation, commonly tested with
MacConkey agar:
1. Lactose fermenters
2. Lactose nonfermenters
 ENTEROBACTERIACEAE

Lactose Non-Lactose
Fermenters Fermenters
No acid
Neutral Red
Acid Colourless
Colonies

Pink
Colonies Shigella, Salmonella,
Proteus, Yersinia
Escherichia, Enterobacter,
Citrobacter, Klebsiella
 General Properties
Members of the family Enterobacteriaceae have the
following characteristics:
– They are gram-negative rods
– Either motile with peritrichous flagella or nonmotile
– Grow on peptone or meat extract media without the
addition of sodium chloride or other supplements
– Grow well on MacConkey agar
– Grow aerobically and anaerobically (are facultative
anaerobes)
– Ferment glucose, often with gas production
– are catalase positive
– Oxidase negative (except for Plesiomonas)
– Can reduce nitrate to nitrite
– have a 39–59% G + C DNA content.
 General Properties cont…
- Cell contain a The outer membrane
characteristic antigen,
called the lipopolysaccharide
enterobacterial common (LPS) is called the O
antigen (ECA). The ECA is antigen.
a carbohydrate-derived
cell surface antigen Motile strains have protein
present in all members peritrichous flagella, which
of Enterobacteriaceae extend well beyond the cell
family. wall and are called the H
antigen.
Its occurrence is
important in Cell surface polysaccharides
resistance may form a well-defined
of bacterial cells to capsule or an amorphous
environmental slime layer and are termed
conditions, such as
bile salts in the K antigen
the human Antigens of Enterobacteriaceae family
digestive tract.
 Escherichia coli
Escherichia coli live in the human gut and are usually harmless but some are
pathogenic causing diarrhea and other symptoms as a result of ingestion of
contaminated food or water.
• Enteropathogenic E. coli (EPEC). Certain serotypes are commonly found
associated with infant diarrhea.
• Enterotoxigenic E. coli (ETEC) produce diarrhea resembling cholera but much
milder in degree. They also cause "travelers' diarrhea".
• Enteroinvasive E. coli (EIEC ) produce a dysentery (indistinguishable clinically
from shigellosis).
• Enterohemorrhagic E. coli (EHEC). These are usually serotype O157:H7.
These organisms can produce a heamorrhagic colitis (characterized by
bloody and copious diarrhea with few leukocytes in afebrile patients). The
organisms can disseminate into the bloodstream producing systemic
heamolytic-uremic syndrome (heamolytic anemia, thrombocytopenia and
kidney failure) which is often fatal.
• Other Pathotype
– Uropathogenic E. coli: causes the commonest community acquired ("ascending")
urinary tract infection
 Salmonella
• Salmonellosis, the common salmonella infection, is caused by a variety
of serotypes (most commonly S. enteritidis) and is transmitted from
contaminated food (such as poultry and eggs).
• Salmonella infections result in vomiting or diarrhea, sometimes severe.
• S. enteritidis does not have a human reservoir and usually presents as
a gastroenteritis (nausea, vomiting and non-bloody stools).
• The disease is usually self-limiting (2 - 5 days).
• Like Shigella, these organisms invade the epithelium and do not
produce systemic infection.
• Salmonella typhi is transmitted from a human reservoir or in the water
supply (if sanitary conditions are poor) or in contaminated food. It
initially invades the intestinal epithelium.
• The organisms penetrates (usually within the first week) and passes
into the bloodstream where it is disseminated in macrophages.
• Typical features of a systemic bacterial infection are seen. The Vi
(capsular) antigen plays a role in the pathogenesis of typhoid.
Antibiotic therapy is essential.
 ECONOMIC IMPORTANCE
There are numerous applications using
members of Enterobacteriaceae including:
• Biocontrol in agriculture
• Production of numerous recombinant proteins
and non-protein products
• Control of infection diseases
• Anticancer agents
• Biowaste recycling and
• Bioremediation.
ECONOMIC IMPORTANCE: Spoilage of Food and
Drinks
• Enterobacteriaceae have been associated with
the spoilage of a range foods and drinks .
Examples include:
– beer spoilage by Obesumbacterium proteus
– spoilage of fresh cream desserts by Serratia spp. and
other members of the Enterobacteriaceae, which is
characterised by clotting cream (from acid production)
and gas production (Sutherland et al., 1986).
– In shell eggs, Enterobacteriaceae are responsible for
bacterial rots,
with Enterobacter and Proteus associated with causing
black rots and Serratia with red rot