Haemophilus, Actinobacillus, Pasteurella

Question 1

HAEMOPHILUS (were haemophilus)
–> Characteristics
* Gram- _______, ______
* weakly ______
* Originally based on whether requires _____ or blood _____. Now based on genetic relatedness.
* Normally X factor (hemin) or V factor (NAD) coenzyme in electron transport chain for
respiration. Found in UPR, urogenital tract; will not be found in environment.
* obligate parasite of host mucosal surfaces
* each species is host specific
* taxonomy is in state of flux

Answer 1

negative, coccobacillus, fermentative, blood, factors

Question 2

Haemophilus” spp and their host
* Avibacterium (H.) paragallinarum
* Glaesserella (H). parasuis
* Histophilus (H.) somni

Answer 2

Used to be haemophillus, now this.

Question 3

Other species of less importance
* H. influenzae
Meningitis, (leading cause of bacterial meningitis in children under 1 year of age; significantly reduced with vaccine) otitis media, bronchitis, etc.
* H. felis respiratory disease
* H. haemoglobinophilus
causes Neonatal mortality in puppies
* Taylorella equigenitalis
causes Contagious equine metritis (S.E. Asia, Austraila, New Zealand).

Answer 3

Said won’t be focusing on this

Question 4

Answer 4

things in red = important

Question 5

What disease do you think these pigs have?
* A. Atrophic rhinitis
* B. Swine cholera
* C. Glässer’s disease
* D. Erysipelas
* E. I don’t have a clue

Answer 5

C

Question 6

G. (H.) parasuis
* specific for swine
* requires V (NAD) factor only.
* Nonhemolytic; does not produce an exotoxin
* More than 15 serotypes may occur, based on capsule or lipooligosaccharide
(LOS), or are untypable; usually a herd issue, which is why you want to know the serotypes.

Question 7

Pigs that have been infected with PRRS or are from SPF herds are particularly susceptible.
* Usually occurs in young pigs (important cause of nursery mortality; develop early on in development)
** fibrinous inflammation of serous surfaces (e.g. pleura or
joints)
“Purple ears are often a feature. Bacteria can readily be isolated from the blood and lesions at post-mortem examination. Chronic cases may be more difficult to
diagnose and may require P-M to look for joint and heart valve lesions.”

Question 8

Answer 8

Glasser’s Disease (polyserositis)

Question 9

Answer 9

Fibrin on paricardium and liver

Question 10

Answer 10

Blood clot in trachea

Question 11

Answer 11

Pericarditis; differential?

Question 12

Specific Diseases due to G. (H.) parasuis
* Respiratory Disease
* Usually occurs in older pigs
following previous infection
(mycoplasma or virus)
* Chronic pleurisy
* Differential:
* P. multocida, Bordetella,
Actinobacillus spp., others

Question 13

Question 14

G. (H.) parasuis Virulence Properties
* Capsule
* Lipooligosaccharide
* Adherence factors
* Membrane proteins, no known exotoxins

Question 15

*Adherence Factors
* required to colonize the host
* bacteria attach to mucosal epithelial cells
* Usually mediated through pili, LOS, or membrane proteins

Question 16

Lipooligosaccharide
There are no O side chains (polysaccharide) in the LPS of Haemophilus species examined, and therefore it can be called an LOS
* The Lipid A still has endotoxic properties
** Causes inflammation
* Core oligosaccharide is antigenic

Question 17

Compromising Factors
* Viral Infection: associated with respiratory disease
* Stress: associated with Glasser’s disease, but not required as
newborn pigs are already compromised
The disease is predominately the result of HOST INFLAMMATION-trying to remove a pathogen it cannot

Question 18

Immunity to G. (H.) parasuis
* Protection is usually mediated by antibodies to capsule; however,
there are at least 15 capsule types
* Protective antibodies may also be made to LOS, membrane proteins,
and other somatic antigens.
* Humoral immunity most important

Question 19

Vaccination, treatment and prevention
*Bacterins are effective against homologous serotype (and somewhat against
others), but may not be long lasting
* Attenuated live vaccine also available and effective
* Antimicrobials
* Intentional exposure at 3-5 days of age with low dose of less virulent
serotypes (RISKY)
* Protection also through maternal antibodies

Answer 19

Make sure to get correct serotype.
Don’t necessarily need to be long lasting in the case of Glaccier’s disease

Question 20

Avibacterium (H). paragallinarum
* Specific for avian species
* Requires V factor only
* Nonhemolytic

Question 21

Question 22

Specific Diseases due to A. paragallinarum
Fowl Coryza (acute rhinitis)
* inflammation of turbinates and sinus epithelium
* air sacculitis
* primarily a disease of chickens

Question 23

Question 24

Answer 24

Depressed, discharge near beak, painful, hunched over

Question 25

Answer 25

High morbidity, but not mortality

Question 26

Answer 26

Inflammation of air sacs

Question 27

Answer 27

Inflammation, pus

Question 28

Virulence Properties of A. paragallinarum * Capsule * Lipooligosaccharide * Adherence factors * Membrane proteins?, exotoxins not recognized

Question 29

Compromising Factors * Viral or mycoplasma infection * Stress Again, disease is predominately due to host inflammation

Question 30

Immunity to A. paragallinarum * Protection is usually mediated by antibodies to capsule * Protective antibodies may also be made to LOS, membrane proteins, and other somatic antigens. * Humoral immunity most important

Question 31

Vaccination * Bacterins (to serotypes A, B, and C) are effective * Designed to raise antibody to cell components * Antibodies to capsule are protective, but only against 1 serotype - purified capsules nonimmunogenic * Other serotypes may be present

Question 32

Question 33

* What is the most likely etiologic agent of this outbreak? * A. Pasteurella multocida * B. Mannheimia haemolytica * C. Histophilus somni * D. A virus * E. Any or all of the above

Answer 33

C

Question 34

Histophilus somni * Includes Haemophilus somnus, H. agni, and Histophilus ovis * Specific for bovines and sheep * Requires neither X or V factor, but does **require blood and CO2** * Most have yellow pigment * Weakly hemolytic or nonhemolytic * **Colonies are yellow **when picked up with a white swab

Question 35

Answer 35

Need CO2 and yellow = key characteristic

Question 36

* Normal habitat is the ruminant genital and respiratory tracts * Disseminates via the bloodstream and localizes at other sites (brain, heart, joints, etc.) * Reasons for dissemination are not clear * Probably due to inflammation of endothelium (**vasculitis** is a hallmark of this pathogen) * Stress and viral infection are required; primarily a feedlot disease; rarily see it in cattle that live out on passure

Question 37

Specific diseases due to H. somni * First disease documented by this organism = TME (thrombotic meningoencephalitis) * ****Pneumonia (BRDC or shipping fever) ***** Myocarditis (biofilm formation) * Reproductive failure (abortion, infertility) * Arthritis * Bacteremia * Others

Question 38

Answer 38

TME Eyes drooping, inflammed (key of TME) Laying down; unwilling to get up

Question 39

Answer 39

Pneumonia

Question 40

Answer 40

Myocarditis Acute, necrotizing myocarditis; bacteria are in biofilm-like aggregates and adherent to venous endothelium, with fibrin and neutrophils

Question 41

Biofilm formation * H. somni forms a biofilm under most growth conditions and in host tissues. When in a biofilm, bacteria are **more resistant to antibiotics and host defenses**, but induce **less inflammation**

Question 42

Answer 42

Filaments, fibirn material polysa, protein, extracelluar dNA = components of biofilm.

Question 43

Virulence Properties of H. somni * Endotoxin (Lipooligosaccharide) Phase variation of lipooligosaccharide in vivo * Antigenic epitopes change and enable bacteria to evade host immunity; some are identical to host oligosaccharides * In the host, the bacteria take up sialic acid and decorate their surface with sialic acid

Answer 43

Their whole defense mechanism is to hide from the host.

Question 44

Answer 44

short chains; can phase vary. Can resist host defenses; host makes antibodey repsonse to Lipo, organisms cleared and those that have changed their oligo now persist.

Question 45

Virulence factors Produce a very large protein: IgG Fc receptors (immunoglobulin binding protein [IbpA] on bacterium * bacterium binds Fc region of IgG, protecting the bacterium from host defenses (fibrillar network of IbpA) **** (there is a toxic fic motif in IbpA) ****Survives within phagocytic cells** by inhibiting phagosome-lysosome fusion * LOS causes apoptosis of endothelial cells, leading to vasculitis * Adherence factors-many

Question 46

Region within gene of immmunogobluni binding protein that produces a Toxin/Exotoxin (has not be categorized yet) H. somni produces a cytotoxin as part of a motif of the IgG binding protein A. This motif has been shown to be toxic for bovine endothelial cells and macrophages.

Answer 46

Causes vasculities, enabling bacteria to penetrate host BBB ad enter blood stream.

Answer 47

Third image on right = cell that does produce binding protein.

Question 48

Compromising Factors * Infection with virulent, as opposed to “commensal”, isolates * Stress * Previous infection Disease is due to host inflammation

Question 49

Immunity to H. somni * Antibodies to the bacterium are normally present in animals and should be protective, but no direct evidence for this yet; can survive in phagocytic cells *** Virulence is largely due to avoiding the host immune response * Protective components not well defined, and likely require cellular immunity * Antibodies to fic motif protective in passive immunity studies

Question 50

Vaccination * Bacterins are limited in effectiveness due to phase variation and IgG binding proteins; best against TME * An effective vaccine against BRD has not yet been developed. Need an intelligen vaccine that recognizes antigens expressed in the host.

Question 51

Host specificity * Host specificity is mediated by adherence and **iron acquisition** * Host-specific bacteria obtain iron from host transferrin. Species that can only utilize transferrin of one host cannot infect other animals.

Question 52

Diagnosis of Haemophilus spp. *Culture: * Need to isolate from sterile site or in pure culture – **chocolate agar** = blood agar heated to 80 degrees * Most isolates from animals require NAD only **(blood for H. somni)** * Some species may require CO 2 (H. somni, some G. (H.) parasuis)

Question 53

*Serology (not usually used): * Complement fixation test most common * Slide or tube agglutination tests easy to do, but are not very sensitive or specific * others that are more sensitive and specific include ELISA. * Outer membrane protein preps or exopolysaccharide

Question 54

Treatment * Susceptible to most antibiotics * Use least expensive and most appropriate drugs * Antimicrobial resistance can occur; may need susceptibility test

Question 55

Summary * A. paragallinarum - Fowl coryza *Birds only * G. (H.) parasuis - Glasser’s disease, pneumonia, septicemia *PIGS ONLY * H. somni - TME, pneumonia, myocarditis, abortion, arthritis *BOVINES, and occasionally sheep, ONLY