Module 4 (Gram-negative And Facultative Anaerobic Rods) Flashcards
➢ These microbes constitute the major microflora of the gastrointestinal tract
➢ They have the ability to become
opportunistic microbes in almost all
species of animals
➢ Predisposition to septicemic diseases, enterotoxigenic diarrhea and edema disease
Escherichia
Species of Escherichia associated
with animal infections
• E. coli
• Colibacillosis of fowls
• E. coli infection of swine
(Swine Colibacillosis)
• E. coli infection of cattle
(Colibacillosis of Cattle)
• E. coli infection of lambs
(Colibacillosis of lambs)
• E. coli infection of horses
(Colibacillosis of horses)
• E. coli infection of horses
(Colibacillosis of horses)
• E. coli infection of dogs
• E. coli infection of poultry
• E. coli infection of Rabbits
Morphology, staining features
and cellular composition of Escherichia
➢ Gram-negative rods
➢ Non-spore former, some strains are capsulated
➢ Strains are motile by means of peritrichous flagella (H-antigens)
➢ Invasive strains possess capsular polysaccharides (K-antigens)
➢ K antigens inhibit attachment of the pathogen to phagocytic cells and ingestion by host cells
Growth characteristics of Esherichia
• The organism grows on all ordinary media
• Some strains are aerobes while some are facultative anaerobes
• Positive for indole, nitrate and urease
• Does not ferment sorbitol (E. coli 0157: H7)
Reservoir of Infection Escherichia
• Environment infected with fecal
materials
• Carrier animals
Transmission of Escherichia
• Feco-oral route
Strains of Escherichia associated with animal infections
(Diarrhea and Colibacillosis)
- Enterotoxigenic strains (ETEC) - neonatal piglets, and weanlings
- Entero-invasive strains (EIEC) – neonates that received inadequate amount of colostrum
- Enteropathogenic strains (EPEC) – diarrhea in animals with attaching and effacing lesions in the intestinal tract
- Enterohemorrhagic strains (EHEC)- associated with Edema and enterotoxemia and it involves release of a vasotoxin
- Colibacillosis of fowls- debilitation and chronic diarrhea
- Extra-intestinal pathogenic E. coli (ExPEC)
• Marked by debilitation and chronic
diarrhea
• Fulminating septicemia
• Complicated by respiratory infections
• Associated with salpingitis,
synovitis and panophthalmitis
Colibacillosis of fowls
▪ Marked by colonic malabsorption
3 Distinct manifestations
• Neonatal E. coli enteritis (1-4 days old piglets)
• Weanling enteritis (enteritis occurs shortly after weaning, entero-toxemia cases)
• Edema disease (diarrhea, associated with age, change in feed)
E. coli infection of swine
(Swine Colibacillosis)
➢ Enteric disease (white scours) in
calves during the first week life
E. coli infection of cattle
(Colibacillosis of Cattle)
• Enteric form occurs in lambs 2-8
days old
• Caused by the proliferation of
entero-pathogenic, non-invasive
strains of E. coli in the upper small intestines
E. coli infection of lambs
(Colibacillosis of lambs)
• Accounts for approximately 1% of
abortion cases
• Observed in mares and about 25%
in infected foals
E. coli infection of horses
(Colibacillosis of horses)
• Bacteremia implicated in
“fading puppy syndrome”
E. coli infection of dogs
➢E. coli is rarely implicated in avian
diarrheal diseases
E. coli infection of poultry
• Cellulitis accompanied by dysentery and high mortality
E. coli infection of Rabbits
Virulence Factors of Escherichia
➢ Surface structures expressed as O (somatic), K (Capsular), H (flagellar) and F (fimbrial) antigens attack complement cascade
➢ Plasmid- encoded heat-labile (LT) and heat-stable (ST) enterotoxins
➢ Edema disease toxin
➢ Vasotoxin (edema disease)
➢ Lipopolysaccharide from the outer membrane
➢ Adhesins (adherence to target cells of the GI tract)
➢ Siderophores, Shiga-like toxins (verotoxin)
➢ Cytotoxic necrotizing factors
➢ Hemolysin
Laboratory diagnosis of Escherichia
Samples: Stools
Agent identification
➢ Bacterial isolation (MacConkey, Minca medium and E media
➢ Quantification of E. coli in the small intestines
➢ ELISA for ST and LT enterotoxin
➢ DNA probes for genes encoding enterotoxins
➢ PCR specific for E. coli trait
➢ Detection of genes associated with EPEC
Treatment of Escherichia
• Apramycin
• Chloramphenicol
• Spectinomycin
• Gentamycin
• Trimethoprim-sulfamethazole
• Ceftiofur
• Correction of fluid and electrolyte imbalance
Prevention and Control for Escherichia
➢Vaccination of gilts and sows with
fimbrial K88 antigens
➢Administration of heat-inactivated
bacterins
➢Good husbandry practice
• These are pathogens of the
gastrointestinal tract of animals
• The species of the genus cause
some septicemic infections
Salmonella
Species of Salmonella associated with animal infections
➢ S. typhimurium
➢ S. agona
➢ S. dublin
➢ S. anatum
➢ S. worthington
➢ S. newport
➢ S. Abortus-equi
➢ S. Abortus-ovis
➢ S. gallinarum
➢ S. pullorum
➢ S. heidelberg
➢ S. Abortus-suis
➢ S. montevideo
➢ S. Saint-paul
➢ S. panama
➢ S. arizona
Morphology, staining features and
cellular composition of Salmonella
• Gram-negative
• Non-capsulated short rods
• With peritrichous flagella (majority of strains)
• Some carry fimbriae (S. gallinarum and S. pullorum)
• There is one capsular type (Vi, virulence) though most do not possess capsules
• LPS, O-antigens and antigenic determinants on surfaces of flagella (H-antigens) (Kauffman-White serologic classification
Growth characteristics of Salmonella
➢ Aerobic but some are facultative
anaerobes
➢ Use citrate as a carbon source
➢ Produce gas from glucose
Reservoir of infection of Salmonella
• Soil, water, vegetation and animal feeds
• GI tract of warm and cold-blooded animals
• Carrier animals
Transmission of Salmonella
➢ Feco-oral route
➢ Ingestion
➢ Through the yolk, horizontal
transmission and Argasid ticks (Pullorum and fowl typhoid)
Establishment of infection to
Salmonella
Phase I. Colonization of the Intestine
(mediated by adhesins)
Phase II. Invasion of the intestinal epithelium (inflammatory response with PMN influx and diarrhea sets in if immune response wanes)
Phase III. Stimulation of fluid exsorption
Diseases associated with Salmonella
- Salmonellosis in cattle
- Salmonellosis in swine
- Salmonellosis in horses
- Salmonellosis in dogs and cats
- Salmonellosis in sheep and goats
- Salmonellosis of poultry
- Pullorum/Bacillary white diarrhea
- Fowl typhoid
Etiologic agent/s: S. dublin, S. typhimurium, S. montevideo, S. Saint-paul
• Infect young animals (4 to 6 months old)
• Feedlot cattle are susceptible
• Septicemic in nature or limited to the GI tract
• Pneumonia is acquired hematogenously
• Abortion is common
• Decreased in milk production
• Foul smelling diarrhea
• Fever, depression, anorexia and weakness
• Death of calves in a day or two
Salmonellosis in cattle
Etiologic agent: S.Cholera-suis,S.typhimurium
• Acute, fulminating septicemia or chronic debilitating disease in feeder pigs
• Stress-associated infection
• Purplish areas on the ears, rump and abdomen
• High fever and anorexia
• Postmortem lesions (petechiae, pneumonia, thickening and necrosis of the intestinal mucosa
Salmonellosis in swine
Etiologic agent: S. Abortus-equi, S. typhimurium, S. heidelberg)
➢ Diarrhea with severe abdominal pain, septicemia and colic
Salmonellosis in horses
Etiologic agent/s: S. typhimurium,
S. panama
• Very uncommon
• Due to noso-comial infection
• Abortion, stillbirths
Salmonellosis in dogs and cats
Etiologic agent/s: S. abortus ovis, S. Dublin, S. montevideo, S. typhimurium
Salmonellosis in sheep and goats
Etiologic agent: S. pullorum, S. typhimurium, S. anatum, S. Newport
➢Fatal among chicks (first 2 wks of life)
➢Infects the embryo before egg is hatched
➢Associated with the failure of eggs to hatch
➢Death is due to septicemia
Pullorum/Bacillary white diarrhea
Etiologic agent: S. gallinarum
➢Acute septicemic disease of adult chickens
➢Paresis, anemia, wing drooping
Fowl typhoid
Clinical signs of salmonellosis in
Poultry
➢ Diarrhea, Septicemia, Weakness
➢ Wing drooping
➢ hyperexcitability
➢ Drowsiness
➢ Paresis and joint swellings
➢ Anemia and Hemorrhages
➢ Leukocytosis
➢ Multiple small necrotic area
➢ Enlarged spleen (PM)
Other disease entities caused by
Salmonella
- Paratyphoid (Infectious enteritis of pigs)
Etiologic agent: S. typhimurium - Avian Arizoonosis
Etiologic agent: S. arizonae - Paracolon infection of turkeys
➢maintained in turkey flock via infected eggs
➢Have R plasmids
Virulence Factors of Salmonella
➢ O (somatic) Antigen
➢ H (flagellar) Antigen
➢ Vi (Virulence) Antigen
➢ Invasins (sip, Salmonella invasion protein in pathogenicity islands in the chromosomes)
➢ Adhesins (fimbria encoded genes, fim; plasmid encoded
fimbriae, pef; and long polar fimbriae, lpf)
➢ Toxins (LT-like toxin with phospholipase A activity)
➢ Siderophores (removes iron from iron-binding proteins of host)
➢ Salmonella virulence plasmids (svp)
➢ RNA polymerase containing RPOs that regulate acid tolerance (survival at < 5.0 pH) LPS and Salmolysin (Sly A)
Laboratory diagnosis of Salmonella
Samples: Feces, intestinal scrapings, milk of infected cows, blood samples, spleen and bone marrow)
Preferred culture media
➢Bacterial isolation (MacConkey Agar, XLD agar, Hektoen Enteric Media, Brilliant Green Agar)
Agent
Identification
➢Determination of somatic and flagellar antigens
➢Salmonella-specific DNA probes and primers for PCR
Treatment for Salmonella
➢ Ampicillin
➢ Cirofloxacin
➢ Enrofloxacin
➢ Amikacin
Prevention and Control for Salmonella
➢Elimination of carriers
➢Proper hygiene and sanitation
➢No effective vaccines
➢ Large, non-motile bacteria
➢ Produce sticky colonies when plated on nutrient media
➢ Normal flora of the vestibule, urethra and clitoris
➢ Frequently encountered in infections of the respiratory, intestinal and urogenital tracts
KLEBSIELLA
Species of Klebsiella associated with animal infections
• K. pneumonia (Friedlander’s bacillus)
• K. ozanae
• K. rhinoscleromatis
Morphology, staining features and
cellular composition of Klebsiella
• Gram-negative short plump rods with rounded ends
• Encapsulated and non-motile
• May occur singly or in pairs
Growth characteristics of Klebsiella
• Grow well on ordinary media
• Ferment a number of carbohydrates
• H2S and urease producer
• Nitrites are produced from nitrates
Reservoir of Infection Klebsiella
• Respiratory excretions
• Secretions in animals and man
Transmission of Klebsiella
• Airborne
• Oral route
• Direct contact with carriers
Diseases associated with Klebsiella
- Vaginal discharges
- Metritis, infertility and abortion
- Pneumonia in sheep and goats
- Normal flora of the vestibule, urethra and clitoris
- Coliform mastitis (cattle and sow)
Etiologic agent: K. pneumonia
➢Hyperemia
➢Edema
➢Swelling of the udder
➢Reduced milk secretion
Coliform mastitis (cattle and sow)
Virulence factors of Klebsiella
• Capsule
• Klebecins (bacteriocin)
Laboratory diagnosis of Klebsiella
Samples: Nasal secretions,Milk samples, Blood and Sputum
Preferred culture media: TSA, McConkey agar, EMB and Blood Agar
Agent Identification
➢Bacterial isolation
➢PCR of a target gene
➢Serology (determines the type of antigens)
Treatment for Klebsiella
➢Cephalexin
➢Kanamycin
➢Neomycin
➢Gentamycin
➢Steroids to correct acute
inflammatory symptoms
Control and Prevention for Klebsiella
➢Strict hygiene and sanitation
➢Elimination of carriers
• Inhabitants of the intestines of
mammals
ENTEROBACTER
Species associated with animal
infections Enterobacter
• E. agglomerans (soil-borne)
• E. aerogenes (bovine strain)
Morphology, staining features and
cellular composition of Enterobacter
• Gram-negative
• Small, motile organisms (peritrichous flagella)
• Non-spore forming rod
Growth characteristics of Enterobacter
• Grow in ordinary media
• Facultative anaerobes
• Reduces nitrates to nitrites
• Ferments carbohydrates (glucose with acid and gas)
• Acetate as source of carbon
Reservoir of Infection Enterobacter
• Animal excretions (urine, pus, feces)
• Inanimate materials (soil, sewage, water)
Transmission of Enterobacter
• Immediate contact
• Oral route
Diseases associated with
Enterobacter
• Septicemic conditions
• Enteric infections
Virulence Factors of Enterobacter
➢Adhesins
➢Colonizing factors
Laboratory diagnosis of Enterobacter
Samples: feces
Preferred culture media: MacConkey,Trypticase
Soy Agar, Brilliant Green Agar
Agent Identification
➢ Bacterial isolation
➢ PCR of genes
Treatment for Enterobacter
• Penicillin
• Tetracycline
Prevention and Control for Entrobacter
• Strict sanitation
• Elimination of carriers
• Inhabit the soil, infect water,
intestinal and urinary tracts, blood,
soft tissues and lower respiratory
tracts of animals
• Motile and have a tendency to
swarm in laboratory media
(progressive surface spreading
ability)
PROTEUS
Species of Proteus associated with
animal infections
• P. mirabilis
• P. rettgerii
• P. vulgaris
Morphology, staining features and
cellular composition of Proteus
➢ Gram-negative straight rods
➢ Highly motile (peritrichous flagella)
➢ Produce a swarming growth on agar
Growth characteristics of Proteus
• Ferments glucose, xylose and
sucrose
• Produces urease and hydrogen
sulfide
• Produces indole but some do not (P. mirabilis)
Reservoir of Infection Proteus
• Urine
• Feces
Transmission of Proteus
• Ingestion of infected material
Diseases associated with Proteus
• Urinary and GI infections
(P. mirabilis-infected spayed bitches)
• Otitis externa (dogs)
• Diarrhea (dogs)
Other pathologic entities associated with Proteus
• Wound infection in a dog
• Otitis externa of Dogs and Cats
• Septicemia in dogs
• Urinary and GI tract infection in dogs and cats
Virulence Factors of Proteus
• Urease (histotoxic effect on the urinary epithelium)
• Proteases
Laboratory diagnosis of Proteus
Samples: Feces, Pus, Urine
Agent Identification
• Bacterial isolation and identification
-Preferred culture media: EMB, MacConkey, TSI
• Urine exam
• PCR
Treatment for Proteus
➢ Cephalosporin
➢ Kanamycin
➢ Penicillin
➢ Streptomycin
Prevention and control for Proteus
➢ Strict hygiene and sanitation
• Organisms are seen frequently as tiny, pleomorphic coccobacilli
• Show bipolar staining in smears
Pasteurella
Species of Pasteurella associated
with animal infections
• P. multocida
• P. hemolytica
• P. pneumotropica
• P. urea
• P. anatipestifer
Morphology, staining features and
cellular composition of Pasteurella
➢ Gram-negative bacteria
➢ Non-motile coccobacillary rods which show bipolar staining
➢ Non-piliated, non-flagellated and non-spore formers
➢ Colonies on blood agar are smooth with a characteristic
semen-like odor
➢ Many strains produce a capsular substance
➢ Capsule contains polysaccharide
➢ Organism produce adhesins and cell wall consists of LPS and proteins
Growth characteristics of Pasteurella
• Grows best in serum and blood
• Facultative anaerobes
• Ferments carbohydrates
Reservoir of Pasteurella
• Mucus membranes of susceptible hosts animals
Transmission of Pasteurella
• Inhalation
• Ingestion
• Through bites and scratch wounds
• By airborne droplets
• Food and water contamination
• Direct or indirect
Diseases associated with Pasteurella
- Hemorrhagic septicemia of cattle and buffaloes
- Pasteurella infection in Chickens
(Fowl Cholera) - Shipping fever
- Snuffles and Rabbit septicemia
Etiologic agent: P. multocida
P. multocida serotype 6:B (Southeast Asia),
P. multocida serotype 6:B (Africa)
P. multocida serotype B2 and E (Philippines)
➢ Clinical Signs
• Fever (41-42C)
• Dysentery
• Edema (pectoral region)
• Cyanosis of the mucous membrane
• Hypersalivation
• Hemorrhages on serous surfaces (PM)
• Bloodstained fluid in the thorax and abdomen (PM)
Hemorrhagic septicemia of cattle and buffaloes
➢ P. multocida
➢ Outbreaks begin in a few birds in apparently healthy flocks
➢ Route of entry: through the eyes and abrasions
➢ Vascular damage leads to hemorrhage
Clinical Signs
• Mucopurulent nasal and ocular discharges (acute cases)
• Rales and diarrhea
• Caseous lesions (chronic cases)
Necropsy findings
• Petechial of the heart
• Swollen spleen
• Reddening of the mucosa of the anterior part of the intestine
Pasteurella infection in Chickens
(Fowl Cholera)
• Fibrinous pleuropneumonia/
bronchopneumonia of cattle and calves handled in stressful conditions
• Respiratory distress and fever
Etiologic agent: P. hemolytica (Manheimia
hemolytica A1)
Shipping fever
• Mucopurulent rhinosinusitis
• P. multocida
Clinical signs
➢Fever
➢Sero-purulent nasal discharge
➢Difficulty in breathing
➢Mucopurulent exudates
➢Difficulty breathing, bronchopneumonia
➢Middle and inner ear infections
Snuffles and Rabbit septicemia
Other diseases associated with P. multocida
- Atrophic rhinitis
➢ Distortion of the turbinate bones, snout and facial structures of swine - Pneumonia, Shipping fever, Swine plaque (cattle, sheep, goats, swine)
➢ Associated with stress factors (transport, harsh climate, castration and dipping)
Clinical signs
➢Severe dyspnea, Fever
➢Soft cough, Nasal discharges
➢Ocular and nasal discharges
➢Diarrhea
- Ulcerative lymphangitis of horse
- Duck septicemic disease (infectious serositis)
- Blue Bag (Mastitis in Sheep)
Virulence Factors of Pasteurella
➢ Heat-labile toxins
➢ Surface envelope antigen
➢ Capsule
➢ Lipopolysaccharide/Endotoxin
➢ Outer membranes proteins
➢ Leukotoxins
➢ Hyaluronidase
➢ Neuraminidase
Laboratory diagnosis of Pasteurella
Samples: Tracheal aspirate, Blood, Tonsils of dead animals, serum, Lungs, spleen, air sacs, material from pneumonic lesions, milk, respiratory secretions
Preferred culture media: Blood agar
Agent identification
➢ Bacterial cultivation
➢ Smears prepared from blood, spleen and tissues of animals with septicemia
➢ PCR of Pasteurella genes
Treatment for Pasteurella
• Penicillin
• Ceftiofur
• Tilmicocin
• Florfenicol
Prevention and Control for Pasteurella
• Reduction of stress
• Routinary immunization
• Strict quarantine measures
• These affect fishes, birds, rodents, primates and domestic
animals
• Implicated in plagues spreading
among people through animals via
fleas.
• Infections are clustered during
warm months affecting handlers of
rabbits, cats and coyotes
Yersinia
Species of Yersinia associated with
animal infections
• Y. ruckeri
• Y. pseudotuberculosis
• Y. pestis
• Y. enterocolitica
• Y. intermedia
• Y. frederiksenii
• Y. kristensenii
• Y. aldovae
• Y. rhodei
• Y. mollaretti
• Y. bercovieri
Morphology, staining features and
cellular composition of Yersinia
➢ Gram-negative coccobacilli
➢ Pleomorphic microbes in chains
➢ Non-capsulated but flagellated (ambient temperatures)
➢ Show bipolar staining
➢ Microbes possess invasion locus proteins (AiL) and invasion (Inv) proteins
Growth characteristics of Yersinia
➢ Grows on ordinary laboratory media (MacConkey)
➢ Ferments glucose
➢ Produces urease
Reservoir Hosts of Yersinia
• Birds and soil
Transmission of Yersinia
• Ingestion of the organism
• Through fleas
• Airborne route
• Oral acquisition (predation, cannibalism and scavenging)
Diseases associated with Yersinia
- Bubonic plaque
- Enteric red mouth (freshwater fishes)
Etiologic agent: Y. pestis
• Characterized by hemorrhagic inflammatory lesions of local lymph nodes (“buboes”)
• Systemic dissemination (septicemia)
• Pneumonia is fatal
• Fleas as vectors (Rat fleas/Xenopsylla cheopis)
Clinical Signs
• Regional lymphadenitis
• CNS involvement
• Fever, depression, anorexia, sneezing, coughing
Bubonic plaque
• Hemorrhagic inflammation of peri-oral subcutis
Enteric red mouth (freshwater fishes)
Virulence factors of Yersinia
• Yersinia adhesin proteins (Yad A) (resist activity of complement)
• Virulence proteins (Plasmid-encoded)
• Yersinia outer membrane proteins
• Yersinia protein kinase (Ypk A) (inhibits phagocytosis and oxidative burst)
• Coagulase
• Exotoxin
Laboratory diagnosis of Yersinia
Samples: Lymph nodes, cerebrospinal fluid, transtracheal aspirates, blood
Agent identification
• Bacterial Cultivation
Preferred culture media: Deoxycholate – citrate medium)
• Serologic tests (hemagglutination, hemagglutination-inhibition)
• Animal inoculation (guinea pigs or mice) die in 3 to 8 days
• PCR for Yersinia genes
Treatment for Yersinia
• Aminoglycosides
• Fluoroquinolones
• Chloramphenicol
• Tetracycline
Control and prevention for Yersinia
• Rat proofing of buildings
• Cyanide fumigation
• Flea elimination and rodent control