Based on the picture:
- The leptospira appear tightly coiled or
LEPTOSPIROSIS
Leptospirosis
- disease caused by a bacteria
- very rampant especially in our country
because we are in the tropics and if there is a
storm and there is a flood, people will get
infected in contaminated flood waters
especially if they have a broken skin or
ingested contaminated food or contaminated
water with this specific type of bacteria.
- it can cause a self-limiting influenza-like
illness or in much more disease and this
leptospirosis is also known as Weil's disease
- the worst thing about this one is that it can
lead or progress to multi-organ failure with
the potential for death
Leptospirosis
- caused by a specific type of bacteria called
Leptospira interrogans
Leptospira interrogans
- it is spiral or corkscrew
- the morphology/physical characteristics of
this bacteria is spiral (kaya sya called
leptospira because of its appearance)
2 clinically significant serovars from this species:
1. Leptospira interrogans serovar Canicola
- from the word "cani", this type of
serovars it infects dogs
2. Leptospira interrogans serovar
Icterohaemorrhagiae
- they are the ones that infect rodents
corkscrew appearance, spiral, thin and they
are flexible spirochetes
- we cannot see them with the naked eye, we
must use special equipment called
microscopes in order to see this bacteria
visibly because this bacteria sizes from 0.1-6
um to 0.1-2 um.
- Aside from their small size, they also have
hook at one end or it can also be both ends.
- What do you think is the significance or
purpose nang hook?
- The function of this hook is for the
attachment of the bacteria to the host
- This specific type of bacteria is best seen in
dark field microscopy just like in
Treponema pallidum.
- As you can see here, black yung background
because we use here a dark field microscope.
- This is the simple illustration of Leptospira
interrogans
- They have two periplasmic subterminal
flagella residing between the outer membrane
and peptidoglycan layer
- It is also known as periplasmic flagella
- Yung PF (periplasmic flagella). The one with
the blue arrow.
 ● Periplasmic flagellum for motility
● 2 important pathogenic serovars from this
species are Canicola and
Icterohaemorrhagiae.
● Reside in alkaline water, alkaline-soil survive
in these area for months or years
○ They survive in this type of
environment.
● Also could be found in urine
- What is the significance of this flagella?
- Just like in other bacteria or parasite,
they will aid in the motility for
moving. Either for swimming or for
crawling
- So etong mga bacteria na to, they
swim in your blood and they also
crawl.
- So the morphology and cell structure of
Leptospira are actually unique.
- The outer membrane (OM), wraps around the
protoplasmic cylinder (PC).
- The components of the protoplasmic cylinder
(PC) are the peptidoglycan (PG), inner
- So this is the Leptospira in kidney tissue
membrane (IN), and the cytoplasm (CP).
- The kidney presented are not sure if it is a
- The blue and white arrow indicates the
kidney for human or rodent
rotational direction of the periplasmic flagella
and the protoplasmic cylinder.
- As you can see yung isa counter
clockwise at yung isa naman ay
clockwise.
- This gives the bacteria the spiral
appearance. Para syang paikot or
spiral shape kase the direction of the
periplasmic flagella and and the
protoplasmic cylinder is opposite
direction. So they tend to give the
bacteria a spiral shape. ➔ if we will stain the bacteria using Gram stain,
as you can observe here in the picture.
LEPTOSPIRA INTERROGANS ➔ What can you see? What is the color of the
bacteria? its color red or color pink, so it's
This is the summary of the Leptospira interrogans.
gram negative bacteria. Meaning, the
● A genus of Leptospira
peptidoglycan is, thick or thin? Thin.
● Gram negative obligate aerobe spirochete
○ In order for them to grow, they need
oxygen
● Finely coiled, thin, motile
○ They are motile because they have a
periplasmic flagella and that flagella is
meant for crawling and swimming.
 that's the time that the bacteria can infect a
human being

➔ How about this one? For this one, we used a

➔ For example, a leptospira interrogans infects a
silver stain of leptospira interrogans and as
rodent especially the serovar
you can observe there are hooks at both ends
icterohaemorrhagiae, it will reside to the
of this bacteria and this specific type of
kidneys or it will colonize the renal specifically
serotype is icterohaemorrhagiae
the renal proximal tubules of the rat or the
➔ Again, this is obligate aerobes and it is called
rodents
icterohaemorrhagiae or interrogans because
➔ When the rat urinates, live bacteria will also be
you can see the question mark. The
excreted into the urine contaminating the
appearance of the bacteria is just like a
environment
question mark.
➔ Now for example, a human host has a broken
MAIN MODES OF TRANSMISSION skin or scrapes or wounds on the feet and it
● Infection is acquired from contact through skin, contacts it has a contact with the contaminated
mucosa/conjunctiva with water or soil water or soil, that's the time that the bacteria
contaminated with the urine of rodents, carriers or will invade the human body
diseased animals in the environment.
➔ How are humans infected by this bacteria? So
the infection is acquired from different routes. It
can be direct or through indirect contact of the
bacteria. Direct contact through skin or mucosa
or conjunctiva with water or soil contaminated
with the urine of rodent carriers or disease
animals in the environment because the most
common carrier are the rodents, especially the
common rat which is called rattus norvegicus.
● Ingestion of contaminated water may also cause
infection. There is no documentation of human to
human transmission..
➔ Next, ingestion of contaminated water may also
cause infect infection but there is no
➔ So aside from that RODENTS, there are also
documentation of human to human
other animals that this bacteria infects such as
transmission. So, meaning if the infected human
domestic animals like dog pig or even cow
host urinates his or her urine cannot be able to
➔ So if these animals urinate, they can contaminate
infect other human it's very rare so once that the
the soil, the water, or the mud and this
skin of a human or the mucosa is contaminated
leptospirosis can survive in alkaline water or
or infected with a contaminated soil or water,
 even in fresh water for up to 16 days and in soil
for almost 24 days meaning almost a month.
➔ And if you are not careful, you will be infected.
For example if a human host aside from a cut or
scratch in the skin these leptospirosis will
penetrate the wound.
● For example, if you are not careful of what you
drink, if the water is contaminated with the
urine of infected domestic animals it can lead
to leptospirosis.
● Especially here in the Philippines, there are
times in which we have typhoons or floods
during the rainy month June or August.
● Last year when the typhoon Maring, went
through our land, in a nearby barangay, there
was a barangay captain (not sure) who died
because of leptospirosis because he got
infected when he was trying to save the
residents in their barangay. According to the
news he has a broken skin but although he
has a broken skin because of his duties and
responsibilities as barangay official, he still
want to save his residence, he went through
the blood without any extra protection but
the consequence of that, he was infected by
leptospira interrogans.
● Even in gardening since this bacteria can
infect the soils, be careful when you are
gardening and you have scrape or cut in your
hands. Make sure to wear gloves, thick gloves.
Because you’ll never know if there are
bacterias present.
● Even swimming in an infected area. What if
you will never know, you have drink the
contaminated water, that can lead to
leptospirosis
HOW LEPTOSPIROSIS IS SPREAD?
SYMPTOMS
LEPTOSPIROSIS AS DEFINED BY WHO
● Leptospirosis is a bacterial disease that
affects both humans and animals. The early
stages of the disease may include high fever,
severe headache, muscle pain, chills, redness
in the eyes, abdominal pain, jaundice,
haemorrhages in skin and mucous
membranes (including pulmonary bleeding),
vomiting, diarrhea and rash.
➔ If you experience this kind of
symptoms, if a family member
experience this kind of symptoms you
may consult a doctor.
LEPTOSPIROSIS SYMPTOMS
● Once that bacteria entered the human body of
course there still a way of our body to fight
these organisms. So this bacteria will go into
the lymphatics and then into our bloodstream
and from our bloodstream, the infection can
spread to the entire body but tends to settle in
the liver and the kidneys .
● This leptospirosis tends to settle or
decolonize in the kidneys and in the liver and
usually it takes around 1- 2 weeks for the
infected person to begin to show symptoms.
● The incubation period of this bacteria is
10-14 days
● For example if you are infected today, he
ingested a contaminated water, after 2 weeks
that is the only time that he/she experience
the signs and symptoms of the disease.
● Fever
➔ Since our body or white blood cells
will try to combat this bacteria
● Intense headache
● Muscle pain or myalgia
 ➔ Muscles are very painful and severely
tender
★ These symptoms are very useful for the
doctors in differentiating leptospirosis from
other diseases causing fever
● Conjunctival suffusion/Redness of the eyes
● Pulmonary manifestations (cough and
chest pain) Jaundice
- Excessive bilirubin in the liver, it will be
SPECTRUM OF ILLNESS stored on the mucosal area of our body.
● This leptospirosis can present in 2 distinct - Nagyeyellow ang mata, skin
clinical syndromes - Because leptospirus colonize in the liver

ANICTERIC ICTERIC
LEPTOSPIROSIS LEPTOSPIROSIS
(Weil’s Disease)

-It is the milder form -It is the severe form of

of disease disease.
-Patients have fever, -Same symptoms with
myalgia but do not anicteric but
have jaundice. characterized by
-Almost 90% of jaundice and is usually
patients have this associated with
type of illness involvement of other
organs. (mas systemic Kidney Failure
na sya, it can lead to - Colonize the kidney
hemorrhage in which
patients may have
spontaneous superficial
bleeding such as
petechiae or purpura )
Petechiae / purpura
- Type of bleeding in the skin
- Or even in GIT
- Leptospires colonizes in GIT or in the liver

NOTES:
- The most common syndrome is ANICTERIC
LEPTOSPIROSIS in which the patient has a
high fever and severe headache that last 3-7
days followed by the immune stage
- Symptoms associated with immune stage
coincides with the appearance of IgM.
Scleral icterus - Once there is an immune stage, there will be a
- Conjunctival suffusion in the presence of production at plasma cells which produce IgM
scleral icterus in order to combat the bacteria.
- Presence of red or bleeding sa may sclera ng - Always remember that the hallmark of the
mata immune stage of leptospirosis is aseptic
 meningitis in which the brain is infected by
the bacteria
ICTERIC LEPTOSPIROSIS (Weil’s Disease)
- Most severe.
- Leads to lethal pulmonary hemorrhage.
- Death can occur in up to 10 % of cases.
How does our body defend or how does our
immune system respond to this disease?
HOST DEFENSE
- ●
- Adoptive immunity-plays a significant role.
- 1. B cell activation with the help of Th cells
through L. interrogans proteins
- Primary host defense mechanisms:
❖ Opsonin- mediated phagocytosis : C3b
fragment and IgG antibodies bind to
the Leptospires cell surface, interact
with receptors on the phagocyte and
initiate
phagocytosis.
❖ complement mediated lysis : classical
complement activation with Ig M and
IgG
● 2. Role of cellular immunity – is controversial.
Experimental mice depleted of T cells have
more severe lung and kidney damage in
response to infection.
So, what is the first thing that will happen if the
T-helper cell will recognize the pathogenic
microorganism which is the leptospira interogans
proteins?
The T-helper will present the antigen to the
B-cells.
Now, yung mga B-cells na yun will become
activated and they will differentiate into
what? What type of cell?--- they will be
activated and differentiate and become
plasma cells
What is the role of plasma cells?
● They will secrete antibodies (IgM,IgG)
this antibodies they will bind to the
leptospira cell surface
● And they will also interact with
receptors on the phagocyte and also
initiate phagocytosis
● Another host defense mechanism is through
complement mediated lysis, by the use of
classical complement activation.
● Remember, the classical complement
activation with IgM and IgG, so if there is a
antigen to antibody complex, the C1 complex
will combine the FC portion of the antibody.
Now, when that happens, the classical
pathway will begin from C1, QRS, then
becomes C3 convertase, to C5 convertase,
which will clevege C5to C5B and then the C5B
will initiate MAC or membrane attack
complex
● what is the end result of classical
complement activation? CELL LYSIS
So those are the two defense mechanism in adaptive
immunity.
First by the use of activation of B-cells that will
differentiate into plasma cells and this plasma cell
will secrete IgG antibodies and aside from that, once
the complements are activated it will lead to cell lysis.
2. The role of cellular immunity is controversial.
Experimental mice depleted of Tcells have more
severe lung and kidney damage in response to
infection.
● So hindi pa masyadong clear according
immunity, but for now we have to focus on the
adaptive immunity of our host.
DIAGNOSIS
Difficult to diagnose clinically; laboratory support is
indispensable.
Supportive
 ● Leptospira agglutination titer of > 200 but <
800 by Microscopic Agglutination Test (MAT)
in one or more serum specimens, or
● Demonstration of anti-Leptospina antibodies
in a clinical specimen by indirect
immunofluorescence, or
● Demonstration of Leptospira in a clinical
specimen by dark-field microscopy, or
● Detection of IgM antibodies against
Leptospira in an in acute phase serum
specimen.
Now how are we going to diagnose the patient, what
are the serologic test in order to diagnose the patient
if he/she is suffering from disease.
So here, the laboratory testsor serological tests that
can be done.
● First, is the use of MAT or microscopic
agglutination test in which titer of greater
200, but less than 800 in one or more serum
specimen.
● So the specimen for MAT is serum.
● What else?
● Demonstration of anti-leptospira antibidoies
in a clinical specimens by indirect
immunofluorescence or by detection of
antibodies, what else?
● Demonstration of leptospira in a clinical
specimen by dark field microscopy or by
detection of IgM antibodies against leptospira
in a acute phase serum specimen.
LABORATORY DIAGNOSIS
Serological Tests
Both are restricted to well equipped expert labs
● MICROSCOPIC AGGLUTINATION TEST (MAT)
- Gold or reference standard
- Requires panels of locally representative
serovars
- May provide indication for infecting
serogroup
- Detects antibodies to specific serovars using
live leptospiral antigens
- Specimen: serum
- Include serovars icterohaemorrhagiae and
canicola
- Interpretation: less than 100 NEGATIVE;
less than 800 POSITIVE (suggested of
previous infection); greater than 1,600
RECENT INFECTION (in the absent of
vaccination)
● ELISA
- The most commonly used method
- Detect IgM antibodies against leptospires
- Detectable during 1st week of illness
because during the incubation, our
body/plasma cells will secrete
antibodies. The first one to fight will
be IgM antibodies
- Genus specific
- In-house ELISA: dilution titers
- Testing in-house ELISA with formalin,
treated and ___ bacteria from the
intermediate species leptospira as an
antigen to detect leptospira specific
IgM antibodies
- Well is coated with leptospira antigens
→ add sample to the sample diluent
→ incubate 30 mins 37 C
→ if antibodies are present it will bind to
leptospira antigens
→ wash then add enzyme conjugate
→ incubate 30 mins 37 C
→ wash to remove unbound antibodies
→ add TMB substrate A+B
→ incubate 10 mins at RT
→ using special equipment to count the titer
of antibodies present in the serum
● LEPTO LATERAL FLOW IMMUNOASSAY
➔ One step colloidal gold
immunoassay to detect antibodies
specific for Leptospira antigens
using a nitrocellulose platform.
➔ A mobile detection reagent consisting
of red, colloidal gold particles onto
which anti-IgM-antibodies have
been bound serves to capture the IgM
antibodies in the patient’s sample
(blood or serum). All the patient’s IgM
antibodies present in the clinical
sample will be captured by the
colloidal gold particles and migrate
toward the test zone, which contains
immobilized Leptospira antigens.
When the complexes reach the test
zone, only those complexes that have
antibodies specific for complexes will
continue to migrate to the control site,
where they will be immobilized. This
control site detects the (anti-IgM-IgM)
complexes.
➔ In this test kit (photo above), present in the
conjugate area the:
- gold antibody (if this antibody is
present, it will bind to the gold
antibody)
- In the test line we have the antigens
of Leptospira.
- And this complex (yung may red dot sa
pic ) will be trap or captured in the
test line. The excess conjugate will
bind to the control line. That gives a
positive reaction.
➔ In the photo below, we use 10 microliters of
the sample and the add 2-3 drops of the
buffer and wait for 15 mins.
● LEPTO DRI DOT AGGLUTINATION
➔ Once there is an agglutination that is
observed, it means it is positive. If
there is none, it is negative or
non-reactive.
➔ An agglutination test that uses
colored latex particles activated
with Leptospira antigen to detect
antibodies specific for the antigen.
➔ In the presence of specific antibodies,
a fine granular agglutination is
observed.
➔ In the absence of specific antibodies (
a negative sample) the blue
suspension remains homogeneous.
● LEPTOTEK DRI DOT
➔ Standard method for diagnosis of
leptospirosis is the microscopic
agglutination test (MAT)
➔ MAT is laborious and time-consuming,
interpretation of results is subjective.
➔ Principle:
- Leptotek Dri Dot assay:
colored latex particles
activated with broadly
reactive Lesptospira antigen
died onton an agglutination
card
 - Assay based on binding of
Lesptospira-specific
antibodies, present in serum
sample, to Leptospira antigen
causing a fine granular
agglutination at edge of
droplet.

TREATMENT:
● Penicillin G -> drug of choice -> 6 million
units daily I/V
● Other drugs -> Ampicillin, Amoxicillin, and
Doxycycline
● In more severe cases -> Cefotaxime or
Ceftriaxone
● Glucose and salt solution infusions
● Dialysis in serious cases
● Organ specific care and treatment are
essential in cases of renal, liver, or heart
involvement.