Bacteriology bacteria. .Campylobacter.ppt

Bacteriology

1. Species Host (s) Disease
C. fetus subsp. venerealisCattle Epizootic bovine infertility
(Bovine genital campylobacteriosis)
infertility, early embryonic death and
occasional abortion
C. fetus subsp. fetus
(C. fetus intestinalis)
Sheep
Cattle
Abortion
(Ovine genital campylobacteriosis)
Occasional abortion
C. jejuni Cattle
Sheep
Poultry
Dogs and cats
Winter dysentery & Mastitis
Abortion
Avian vibrionic hepatitis (Helicobacter
pullorum)
Enteritis with diarrhoea
C. mucosalis Pigs Swine proliferative enteritis
(Porcine intestinal adenomatosis complex)
C. hyointestinalis
(Serpulina
hyointestinalis)
Pigs Swine proliferative enteritis
(Porcine intestinal adenomatosis complex)
C.coli Pigs, Man Mild diarrhoea in pigs and enteritis in
man. Swine dysentery
(along with Serpulina hyodysenteriae)
Campylobacter

2. Habitat and Ecology
worldwide in distribution
commensals on the mucosa of the oral
cavity and intestinal tract
C. fetus subsp. venerealis - prepuce of
bulls and
in the genital tract of cows
Some nonpathogenic species that

are saprophytes in the environment.
Obligate parasite.
C. jejuni - natural avian reservoir

3. Morphology
Gram negative, slender, short curved rigid rods - tends to
be pleomorphic
loosely attached capsular envelope, referred to as
the S – layer
Arranged singly comma-shaped or united to form ‘S’
shaped , sea gull shaped or spiral shaped

4.  Motile - Long sheathed polar flagella at one (polar) or both
(bipolar) ends of the cell
 Wet mounts under phase contrast or dark field microscopy
- curved forms with distinctive rapid darting motility or
Cork-screw motility.

5. Cultural characteristics
Microaerophilic - reduced oxygen & 3-15 % C02
Optimum temp- 37
o
C. Some grow at 42
º
C
thermophilic (42-43
º
C)
Non-hemolytic.
Slow growing organisms
very fastidious
Blood agar with or without thioglycollate
Serum-dextrose agar
Thiol medium
Clark and Dufty medium
Chocolate agar
Skirrow's and Bultzler's medium
Microaerophilic

6. Growth on Skirrow's and Bultzler's medium
Grown on blood free, charcoal based
media
Charcoal-cefopera-zole-deoxycholate
agar or Blaser's Campy-BAP medium-
Brucella agar with 5 per cent sheep blood and vancomycin
(10 ug/ml), polymyxin B sulphate (2-5units/ml),
trimethoprim lactate (5 ug/ml) cephalothin (15 ug/ml), and
amphotericin B (2 ug/ml).

7. Cultural characteristics
Colonies - small, round, slightly raised, smooth,
translucent colonies 'dewdrop' appearance.
Colonies fuse together - produce a large mucoid
colony
Surface of the colonies - frosted appearance
Organisms without capsule – rough colonies.
C. coli produces a pink-tan pigment
C. mucosalis -dirty-yellow pigment.

8. Bio-chemical properties
Do not ferment carbohydrates
catalase positive, oxidase positive, nitrate reduction and
growth in 1% bile.
C. jejuni is the only species that hydrolyses sodium
hippurate.
Resistant to nalidixic acid but susceptible to cephalothin.
A large loopful of a 24-48 hour culture of C.
jejuni is emulsified in 0.4 ml of 1 % aqueous
solution of sodium hippurate and incubated at
37°C for 2 hours.
Add 0.2 ml of a ninhydrin solution at 37°C. A
positive reaction is given by a deep purple
colour developing after 10 minutes.

9. Resistance
Fragile, not withstand drying &sunshine,
readily killed by disinfectants.
Killed at 60
o
C for 5 minutes.
Survive in soil, hay and manure for 10-20
days depending upon the humidity and
temperature.

10. Enterotoxin
Endotoxin
Adhesins
Ability to penetrate cells
Virulence factors

11.  Present in penis and prepuce of bulls and genitalia of cow
lower breeding efficiency.
Bovine infertility
Endometritis
Failure to implant or early abortions.
Abortions at
  5 - 7 months
Bull - inapparent infections of penis and prepuce.
 Self-limiting:
  eliminated in 5 months.
Campylobacter fetus subsp. venerealis
usually through coitus or artificial insemination
The organism localizes in the anterior vagina and
cervix during the ovulatory phase but does not invade
the uterus and oviducts until progesterone release - and
then causes endometritis and salpingitis for several
weeks to a few months during which the animal in
infertile - animals usually regain fertility within 5 months.

12. show no initial clinical signs until the fetus is aborted,
Lesions in aborting ewes include
swollen caruncles with exudate
thickened uterine walls
placentas with mottled
swollen cotyledons
 Aborted fetuses are usually autolyzed, with fluid in the
abdomen and thorax
Liver necrosis, suppurative bronchopneumonia -Characteristic
necrotic spots in the fetal liver
Campylobacter can be cultured in large numbers from
aborted placentas
Campylobacter fetus subsp. fetus
Organism enters via ingestion, (venereal transmission does
not occur) causes bacteremia and finally localizes in the
placenta causing placentitis and abortion near the end of
gestation

13.  Zoonotic pathogen.
 Normal flora of most domestic and wild animals,
Common contaminant of streams and fresh water.
Transmission to humans via milk, meat, dogs, cats, and poultry.
Implicated as the etiology of "Winter dysentery of cattle”
Adult cattle are most susceptible.
Diarrhea in calves, dogs, cats, and foals,
Bovine mastitis.
Enteritis in humans.
Milk, minced meat, dogs, cats, and poultry are important
reservoirs of infection for humans
In human even 500 c.f.u./ml in milk are infective, the signs are
fever, abdominal pain, nausea, vomiting, and bloody stools. Some
may become persistently infected.
Campylobacter jejuni

14. Specimen: Vaginal mucus & preputial washings
Vaginal mucus three weeks after AI or service - most ideal
material.
Foetal abomasal contents using the DCF stain.
Based on symptoms, lesion and history of abortion in a
herd.
Microscopical examination of smears from foetal stomach
contents or visceral organs - by Gram’s staining method -
characteristic morphology.
Isolation of the bacteria by culturing: pinpoint, delicate,
circular and opaque colonies in selective medias.
Vaginal mucus / Cervical mucus agglutination test : A
positive reaction is at least 75% agglutination in the first two
tubes
Diagnosis

15. Species Host (s) Disease
Dichelobacter
(Bacteroides) nodosus
Sheep
Cattle, Goats and
Pigs
Contagious (virulent) foot rot
Contagious interdigital dermatitis
B. melaninogenicusCattle Foot rot
B. asaccharolyticusDogs, cats, horses,
cattle
Osteomyelitis
B. fragilis Calves, lambs, foals,
piglets
Diarrhoeal disease
Fusobacterium
necrophorum
(associated with
Corynebacterium
pyogenes species)
Sheep
Cattle
Horses
Pigs
Chicken
Foot abscess
Ovine interdigital dermatitis ( ‘Scald’)
Foot rot, bovine liver abscesses, and Calf
diptheria
Thrush - infections of front of hoof.
Ulcerative stomatitis
("sore mouth" or "Bull nose" - via injury
from fitting nose rings)
Necrotic enteritis
Avian diptheria
(Secondary to fowl pox)
Bacteroides and Fusobacterium

16. B. nodosus is an obligate anaerobic bacteria of
the digital epidermis of sheep under normal
climatic conditions.
Fusobacterium species ubiquitous in soil and
manure, in the gastrointestinal tract, and on the
skin and hooves of domestic animals
Habitat

17. Bacteroides nodosus appear
as Gram-negative, fairly large
(1.7 x 3-6 urn), slightly curved and non-
motile rods. Often swollen at one or
both ends. occur singly or
occasionally in pairs-
(barbell or club shaped)
Fusobacterium necrophorum
is Gram-negative, long and
filamentous but does not branch.
Filaments can be up to 100 µm in
length and 0.5-0.7 µm in diameter. May
have tapered or rounded ends.
Irregular staining is characteristic
Morpholo
gy

19. Anaerobic jars with a catalyst, an anaerobic
indicator and an atmosphere free of oxygen
(10%
Hydrogen, 5% Co2 and 85% Nitrogen) is
essential
Cultural
characters

20. Eugon agar base with 0.2 % (w/v) yeast extract,
10 %defibrinated horse blood and one µg/ml
lincomycin is the selective medium for Bacteroides
nodosus.
Lemco agar containing pulverized hoof powder
Agar media should be pre-reduced in an anaerobic
jar for 6-24 hours before use. Plates are incubated at
35-37°C immediately after streaking for 4-8 days.
Liquid media such as Cooked meat broth with 0.4
per cent glucose or thioglycollate medium is also
suitable with the addition of the vitamin K-haemin
supplement.
Cultural
characters

21. Colonial morphology
Bacteroides nodosus
Three basic colonial types
B-type: papillate or beaded (most pathogenic) from ovine foot rot.
M-type: mucoid (less pathogenic) from non-invasive infections of
sheep and cattle.
C-type: circular (non-pathogenic) and resulting from repeated
passage in media.
Greyish-white in color
Bacteroides melaninogenicus
 Circular, entire, convex and shiny colonies, 0.5- 2.0 mm in

diameter. Colonies become darker after 5-14 days, being black in
the centre with a grey-brown periphery.
Haematin (black or brown) pigment is seen best on media
containing blood.
 A few strains are haemolytic on rabbit blood agar.

23. Fusobacterium necrophorum
F. necrophorum produces grey to yellowish shiny
colonies on blood agar, that are about 2-3 mm in
diameter after 48 hours incubation. Haemolysis is
variable.
Gram-stained smear from the colonies shows
long Gram-negative filaments that are less
characteristic than those from direct microscopic
examination of specimens.
Lipase, but not lecithinase, activity is exhibited by
many strains of F. necrophorum on egg yolk agar.
Colonial morphology

25. All pathogenic Bacteroides species are
catalase negative except B. fragilis.
B. nodosus does not ferment carbohydrates.
F. necrophorum is indole and H2S positive.
Gelatin is not liquefied.
Ferment glucose and maltose with
production of acid and gas but not lactose.
Biochemical characters

26.  B. nodosus causing foot rot produces
keratinolytic enzymes in greater amounts
 F. necrophorum produces necrotizing
endotoxin and exotoxins leukotoxin, hemolysin,
hemagglutinin, proteases, and adhesin
Endotoxin when inoculated intradermally into
rabbit causes necrosis.
Exotoxin when inoculated intradermally into
rabbit causes mild erythema but when
administerd intravenously causes emaciation or
death of rabbits in a few hours.
Inoculate rabbit subcutaneously with
F. necrophorum produces lesions throughout
the body
Toxins

27. Foot rot is a contagious, acute or chronic dermatitis involving
the hoof and underlying tissues
Cattle
In foot rot – acute painful swelling of a hoof which leads to
lameness.
Necrosis in digits spreads to tendons, ligaments and joints.
Sheep:
 
Foot rot (interdigital dermatitis, infective bulbar
necrosis and heel abscess), mouth lesions and abortions
(rare).

31. Swine: 
Principal cause of bull nose or Ulcerative
stomatitis via injury from fitting nose rings.
Cats: 
Opportunistic, highly suppurative.  Involves
nasal passages, oral cavity and bone. Secondary
invaders to tissue damage. Dental tartar leads to
gingivitis and periodontal disease.

32.  
 Liver abscesses arise with the organisms that inhabit
the rumen gaining entry into the portal circulation, and
are often secondary to ruminal acidosis and rumenitis
complex in grain-fed cattle.
 
Tissue necrosis, as well as the foul-smelling waste
gases produced by the bacteria, are characteristic of
necrobacillosis, or
 
F. necrophorum 
infection.
Clinical signs include necrotic, fetid lesions, usually of
the mouth or feet, that can cause ingestion or lameness
problems
Calf diphtheria is more common in 3 to 18 months old.
Malignant form of infection. More frequently found in
bucket fed calves compared to suckling or bottle fed
calves.
Cattle
 

33. Necrobacillosis

35. Human- Sore throat – F. necrophorum

36. Collection of specimens
 Specimens should be placed immediately in an oxygen-
free container, especially small pieces of tissue or material taken
on swabs.
Larger pieces of tissue (over 2 cm
3
) usually maintain an
anaerobic microenvironment deep in the tissue and can be placed
in an air-tight jar for transportation.
Fluid specimens can be collected in a sterile syringe, the air
expelled and the needle bent over or plugged. However, if the
specimen cannot be processed within an hour, a fluid
specimen should be placed in an oxygen free tube or vial.
All specimens for anaerobic culture should be processed
within a few hours of collection. It is best to keep the specimens at
ambient temperature rather than in the refrigerator, as oxygen
absorption is greater at lower temperatures
Diagnosis

37. 1. Direct examination
Fusobacterium necrophorum in clinical specimens is long and
filamentous (about 1m in diameter) and characteristically
stains inan irregular manner.
Bacteroides nodosus is a large rod characterised by the presence
of terminal enlargements at one or both ends (barbell or club
shaped). FAT is reported as being specific and sensi
­tive.
2. By Isolation and identification in anaerobic media
3. By animal inoculation
Mice, rats and guinea pigs inoculated with cutlrues of B. nodosus
will produce abscess and sepsis.
Inoculate rabbit subcutaneously with material suspected for
F. necrophorum produces lesions throughout the body.
Differential diagnosis
Foot abscesses, selenium/vitamin E deficiencies, strawberry foot
rot,
 
bluetongue virus 
infection
 FMD
Pyogenic infections associated with Corynebacterium pyogens.
Diagnosis

38. Surgical paring of hoof to remove as much dead necrotic
material as possible to expose the hoof to oxygen.
Topical application of 10% formalin or Copper sulfate or
Zinc sulfate foot bath are effective in reducing the incidence
in the herd.
In B. nodosus infection the affected hooves should be
trimmed and treated with 10% formalin or chloramphenicol
or tetracycline. Parental treatment with penicillin and
streptomycin may be of value.
 In Fusobacterium infections, 5-10% CuSO4 is
recommended for the treatment of foot lesions. For early
treatment sulphonamides, tetracycline and erythromycin are
useful.
Control and
treatment