TOPIC:

SPIROCHETES
Submitted to
Dr. Rokeya Ahmed
Asst. Lecturer of Microbiology
Faculty of Veterinary & Animal sciences
Gono Bishwabidyalay Submitted by
Md. Zahid Hasan(232)
Shuvenkar Dey(233)
Md. Raju Mola(234)
Md. Mosgul Mia(235)
Md. Rakibul Hasan(236)
Samia Jahan Shova(237)
Tajin Islam Priyanka(238)
Faculty of Veterinary & Animal sciences
Gono Bishwabidyalay
Spirochetes
Spirochetes
Family
_____________________
C
V
Spirochaetanceae Leptospiraceae
G Genera V
Borrelia; Leptospira
Treponema
• Majority are saprophytes ,a few are commensals and some are pathogenic ,
causing diseases in both humans and animals.
• Spirochetes are inactive biochemically and hence differentiation is difficult by
means.
Identification is based on antigenic properties:
• Although spirochetes are cyto-chemically Gram negative but they stain poorly
.they may be demonstrated by special procedures---
a) Giemsa or Wright Stain
b) India ink or Nigrosin (Negative stain)
c) Silver Impregnation
d) Darkfield Microscopy
e) Immunofluorescence
 Leptospira
Species:
• Two species—
a. Leptospira interrogans
b. Leptospira biflexa
Leptospira interrogans cause disease in man and
animals whereas Leptospira biflexa is free living
saprophyte. There more than 180 serovars and
serogroups.
Host Species
Cattle Leptospira interrogans serovar pomona
Leptospira interrogans serovar grippotyphosa
Leptospira interrogans serovar canicola
Leptospira interrogans serovar icterohaemorrhagiae
Leptospira interrogans serovar hardia

Horse Leptospira interrogans serovar pomona

Leptospira interrogans serovar canicola
Leptospira interrogans serovar grippotyphosa
Leptospira interrogans serovar icterohaemorrhagiae

Sheep & Goat Leptospira interrogans serovar pomona(sheep)

Leptospira interrogans serovar grippotyphosa(goat)

Pig Leptospira interrogans serovar pomona

Leptospira interrogans serovar canicola
Leptospira interrogans serovar icterohaemorrhagiae

Dog Leptospira interrogans serovar pomona

Leptospira interrogans serovar canicola
Leptospira interrogans serovar icterohaemorrhagiae
Morphology
• Tightly coil , thin , flexible spirochetes
• Closely spiraled with one end commonly bent into
the shape of a hook
• Active rotational motion but no flagellum has
reported.Electon micrographs show athin axial
filament and a delicate membrane.
• Barely be seen by the usual microscope
method s but easily visualized by darkfield
microscope .It does not stain normally with
conventional coalter dyes but can be
imagegnated with Silver salts using a technique
described by Fontana.
•.
Growth characteristics
• Grows best under aerobic conditions at 29°c to 32°c
in protein rich semisolid media (Fletcher’s media
)where they produce round colonies 1 to 3 mm in
diameter in 6 to 10 days.
• Leptospira also grows in chorioallantoic membrane
(CAM)of embryonated eggs.
• Leptospira derive energy from long chain fatty acids
and cannot use amino acids or carbohydrates as a
major source source of energy . Ammonium salts are
main sources of nitrogen .
Mode of infection
• Direct contact with the urine of infected animals or
ingestion of urine contaminated with water or food
.Transmission is even possible through drinking water
contaminated by urine of rodents or wild animals.
• The organisms may enter the animal’s body through
abraded skin , mucous membrane or conjunctiva.
• Transplacental infection can occur in cattle ,pig and men .
• Calves can acquire infection through milk.
.

Fig:Mode of infection of Leptospira

Fig:Mode of infection of Leptospira

Pathogenicity
Bovine Leptospirosis:
• Principally caused by Leptospira interrogans serovar
pomona,hardio causes fewer abortions but results in infertility
.Serovar grippotyphosa ,canicola or icterhaemorrhagiae is
involved occasionally.
• Sources of the organosm are urine .Cattle may shed the organism for
3 months.Outbreaks are associated with haevy rainfall.
• Red water (discolared urine from pink to dark brown ,including acute
nephritis )is seen in calves and young animals accompanied by
anemia and jaundice .Encephalitis may occur . Death from renal or
hepatic failure may occur in 3 to 10 days while survivors may develop
chronic nephritis with poor renal function and excrete leptospirae.An
outcome in pregnant cows is abortion or stillbirth, one to tree weeks
after the acute illness.
Lesions:-
• Jaundiced apperrance of carcass,
• congestion of kidney with pinpoint hemorrhages,
• enlargement of spleen.
Canine leptospirosis:
• Primarily caused by Leptospira interrogans serovar canicola and less
frequently by Leptospira interrogans serovar icterohaemorrhagiae.
• Infected dogs sporadically shed Leptospira in their urine and serve as
sources of leptospira interrogans serovar canicola and Leeptospira
interrogans serovar icterohaemorrhagiae infections & human may be
infected .
• Four principal forms are recognized:
1. The hemorrhagic form
2. The icteric form
3. The uremic or subacute form
4. The inapparent form.
The first two forms are primarily caused by serovar icterohaemorrhagiae
& the latter two forms caused by serovar canicola

Clinical Lesions:
• Vomiting in acute case
• Red eyes progresses to nephritis(Arched back, immobility , blood stained
urine)
• Dehydration
• Death within 4 days
Equine Leptospirossis:
• Congenital infection ,stillbirth .abortion Clinically same like
cattle.
A form of chronic recurrent eye infection (uveitis) may
develop 2 to 8 months later.
Swine Leptospirosis:
• Abortions and stillbirth, congenital infections.
Diagnosis
Specimens :
1. Urine , portions of kidney and liver.
2. Paired serum samples for serological test
Procedure:
1. Microscopic examination:
Demonstration of Leptospira in fresh urine or formalized urine by
Darkfield examination of thick smears stained by Giemsa’s
technique . Thin,bright .coiled bacteria will be seen. Occasionally
show Leptospira in fresh blood from early infection .Microscopic
examination can also be done by silver impregnation technique
described by Fontana.
2.culture:
Whole fresh blood or urine cultured in Fletcher’s semisolid
medium . Growth is slow and culture should be kept for several
weeks(4 weeks) at 25°c to 30°c.
3.Animal inoculation:
• Guinea pig is inoculated intra-peritonially with fresh
plasma or urine . Within a few days, spirochetes become
demonstrable in the peritoneal cavity on the death of the
animal , haemorrhagic lesions with spirochetes are found
in many organs.
4.Serological test:
• Microscopic Agglutination test (MAT)
• Fluorescent Antibody Technique(FAT)
• ELISA
5.PCR(Polymerase chain reaction)
Control & Prevention
• Rotan control
• Treatment of carrier animals
• Protective clothing and Occupational hygiene
Note:In dog, cat & pigs –Vaccines are available suspensions of killed
serovars.The vaccines are given s/c or i/m in two initial doses, one
month apart followed by boosters.
Public Health Significance
• Human beings acquire infections from infected domestic
animals , rodents and contaminated water.The organisms
may also enter through mucous membranes or breaks in
the skin .
• The disease is referred to several names: Weil’s disease
,Fort Bragg fever and Swineherd’s disease
• Various serovars of Leptospira, including canicola
icterohaemorrhagiae and pomona can infect humans
• Veterinarians, slaughterhouse workers and farmers are at
particular risk .The acute from of the human disease is
characterized by febrile jaundice and nephritis.
Borrelia
Species: Borrelia anserina
Synonym: Borrelia gallinerum
Disease : Avian Borreliosis
Host: chicken ducks turkeys and geese
Transmission: by bites of ticks e.g- Argas persicus and other
Clinical sign:
• Acute septicemia
• Fever
• Diarrhoea
• Drowsiness
• Emaciation
Diagnosis:
• Direct examination-Giemsa stained blood film
• Darkfield microscopy
• Can be cultivated in chicken embryo
 Relapsing Fever
A disease is caused by a number of Borrelia spp e.g- Borrelia
rectarentis,occurs sporadically in U.S.
Borrelia persica-Asiatic African tick borne relapsing fever.It is
characterized by febrile attacks during which the organism can
be recovered from blood.
Borrelia vincentii,Borrelia buccale:-Opportunistic pathogen ,
cause ulcerative disease involving the gum and oral cavity.
Vincent angina(Acute sorethroat), ulcerative tonsillitis in dogs
and other mammals.
Borrelia burgdarferi:Lyme disease in man.
Treponema
Species:
• Treponema suis: Preputial infection in pigs
• Treponema hyodyserteriae:Swine dysentery.
• Treponema cuniculi: Rabbit syphilis or Vent disease.It
is a true venereal disease in which lesion consisting of
vesicles and scabs are seen mainly involving the
prepuce ,vagina and perineal region.Thick scaly crusts
persist in the female for months.
Diagonosis:
Darkfield microscopy
Treponema pallidum:Cause exclusive syphilis in human.
Three forms of the disease:-Primary ,Secondary ,Tertiary
lesion.
Primary lesion:occur by sexual contact and subsequent
appear of syndrome characterized by papule which
undergoes lysis and form primary sore of syphilis.
After 3 to 5 weeks secondary lesions appear,characterized
by red maculopapular lesion and moist papule appears the
anogenital region.
After 3 to 5 years tertiary stage begins characterized by the
development of granulomatous lasions in skin,bones and
liver.
Distinguishing features of Borrelia ,Treponema and
Leptospira:
Characteristics Borrelia Treponema Leptospira

Length 8-16µm 5-18µm 6-20µm

Width 0.25-0.3µm 0.09-0.5µm 0.1-0.2µm
Spiral number 4-8,loose 6-14,regular Many ,fine , tight
Ends Taper terminally Pointed , may One or both ends
To fine filaments have terminal have a semilunar
filaments hook
Staining Gram yes No no
Giemsa yes poor poor

Silver Not yes yes

impregnation necessary
Thank You