MCQ questions

In case of ulcerative lymphangitis in horse, the lymph nodes usualy affected are .1
a- Fore legs LN
b- Hind legs UN
c- Neck LN
d-Chest LN
.Lymph nodes when infected with C-pseudotuberculasis in sheep look like 2 -
.a- Onion like
.b- Pear like
.c-Nodular like
.d- None of all
Strauss reaction could be done in .3 -
a- Rabbit
b- Female guinea pig
c- Male guinea pig
d-Rat
Strauss reaction is characterised by .4-
a-nodules in Inguinal lymph nodes
b-orchitis & pus formation in the testis
c-lymphangitis of ingulnal lymph vessels
d-lympha- denitis of inguinal lymph nodes
C.pseudotuberculosis "C.ovis" causes The following in horse .5-
a- Caseus lymphadenitis
b- Ulcerative lymphangitis
c- Epizootic lymphangitis D-Glanders
Cases

Foal died with pneumonia 24 weeks, lung sprcimen turned to bagteriology lab, the isolated .1
colony was macroscopically, small-medium mucaid, round colonies; no hemolysis and note
....... characieristic salmon pink color, gm stain +ve plodmorphic rods, the isolate is
C. Equi (Rhococcus equi) →CAMP (+VE Nitrate (+ve), Catalase (+ve), sugar fermentation (-ve) >
Pus from a chronic infected Sne leg wa urned te bacteriology laboratory .2
Enumerate name of susprcted bacterial isolates .1
 Covis - Actinobacillus (pseudononts Mallei- Histoplasma farciminosum-
Sporothrixschenkii(dimorpie fuogi)
Confirmatory laboratory dirterentlation between them .2
Disease in Horse Cause Morphology Strauss reaction Mallein test

Ulcerative C.ovis G+ve Chinese Ve+ Ve-

Iymplhangitis Letter like

farcy Actinobacillus G-ve bacilli Ve+ Ve+

(cutenousglanders) (pseadomonas)
Mallei
Epizootic Histoplasma oval G -Ve double Ve- Ve-
lymphangitis farciminosum contoured cells
with bud (yeast
like)
Uicerative Sporothrixschenkii Macro and micro- Ve- Ve-
lymphangitis (dimorphic fungi) conidia outside
host
Urine samples with pus sediments turned to your laboratory from cow with arched back and .3
colicy pain, after staining, the M.O. was gm+ve, and Chinese letter appearance
What is the suspected isolate .1
 C. renale group
Laboratory confirmation of this suspected isolate .2
Samples →Mid stream urine
Direct Microscopic examination →short stumpy. (Complete other characters).
Culture characters:
Aerobic and facultative anacrobes .1
Nutrient agar with 10% skimmed milk →creamy yellow pigmented colonies .2
Blood agar →small non hemolvtic colonies .3
CAMP test →+ve (enhance B hemolvsis of S. Aureus) .4
Blochemical characters
 Only glucose is fermented
 Catalase test →+ve
 Urease test →+ve
 Nitrate test → -ve
Mycobacterium (T.B)

• Species of Mycobacterium (T.B) →

1. Typical group →obligate pathogens
a. Tuberculosis group →[MTB (human -monkey), M.bovis (human M.Avium(birds)}
b. Paratuberculosis group →(M.paratuberculosis or M. pseudotuberculosis or M.johnae which
causes (John's disease) in sheep, goat and cattle which leads to emaciation and diarrhea due to
digestive casteated nodules)
c. Leprosy disease →M. Liprae (Hansen' bacilus in human)
2. Atypical group/Saprophytic →Opportunistic pathogens
Typical T.B Atypical T.B

Pathogenic bacteria Saprophytic & commensal bacteria

(Opportunistic bacteria)

ZN +ve ZN +ve

Acid and Alcohol fast Acid fast but NOT alcohol fast

Grow at 37c Grow at room temperature 25 c

Need special media (LUM -EDM) Not need special media

Long incubation period (Slow) Short incubation period (Rapid Grower)

NOT pigment producers PIGMENTS PRODUCERS

General characters of mycobacteria:

1-Special staining (Zhiel Neelsen) →acid fast resist discoloration by acid
M.O. +strong carbol fuchsin →heating wash by acid alchol sol. + methylene blue (counter stain)
→Red (acid fast) against blue background.
2-Slow growth →Long Incubation period (21-45 days)
3-growth medium is poured in screw copped bottles?? Due to long IP to avoid evnooration of the
media
4-sample preparation before culture (sample processing)?? For liquefaction of the sample -for
conc. Of m.o. (l/cellular with intermittent shedding of low amount) - decontamination of samples
(L.n)
5-resist acid but heat sensitive
5-First diagnosis by allergic reaction (tuberculin test)
6-specific lesion (Granuloma formation)
7- No treatment in animals ( only test and slaughter due to chronic progressive nature and
zoonotic nature of disease) but only in human.
• Virulence factors →
1. High lipid content in cell wall (resist phagocytosis) →granuloma
2. Intracellular survival inside macrophage
3. Waxy coat and other TB protiens stimulate cell mediated immunity & delayed hypersensitivity
Pathogenesis (lesion)
1. Route of infection →inhalation (MTB), ingestion skin (bovis)
2. bacillus entry →odema, eosinophil, macrophages
surrounded by fibrin (Exudative lesion) heal or continue to productive.
multinucleated giant cells + epitheliod cells+ fibroblasts + lymphocytes +fibrous capsule+ calcium
(productive lesion)
Still as tubercle or millary tuberculosis

Diagnosis of T.B
A. Field diagnosis →
 Allergic tests (Tuburclin test) for -> MTB and M. bovis: it (depend on cellular immunity) (T.
cells) that develop 30-50 days post infection so these tests can be done 30-50 days post
infection. (delayed hypersensitivity)
 Tuberculin test (accuracy, true positive up to 65%)
it is standard antimortem diagnostic test??? Once it give tve →slaughter of the animal types →1.
old tuberculin
2, PPD****** purified protein derivatives" better than old test & more accurate
PPD →MTB, M.bovis, M.avium →inoculate in glycerol broth →incubate(37C,7week)
→centrifugation →supernatant + trichloroacetic acid "precipitate protein " →purification →0.1 mi.
ID.(skin tail fold)
result : after 72 hrs.
1. Same skin thickness →-ve.
2. skin thickness →4mm →+ve "granuloma formation→+ve →slaughter.
B. Laboratory diagnosis →
1. Samples →
a. Sputum (pulmonary infection), urine, peritoneal or pleural aspirate-> MTB, human.
b. Lymph node (after slaughter), pus, milk, trachea-bronchlal aspirate, feces, semen →M. Bovis,
cattle.
c. Feces - rectal swab -) M.avium - paratuberculosis
2. Samples processing (precautlons of samples handling) →
a. Aim of sample processing →(1. Liquefaction of samples (Sputum or L.N),
2. Concentration of M.O
3. decontamination of samples)
b. Steps →
Specimen processing:-
Importance of NaOH 4% → 1. Decontamination to other m.o. 2. dissolve mucous
3. Direct microscopic examination →staining processed sample by ZN stain.
Shape : bacilli arrangement: single or pair noncapsulated, nonsporulated, non motile, stain: 1.acid
fast "by Zhiel Neelsen" →red bacilli and blue background
2. gram positive
4. Culture characters →
a. Environmental factors →long incubation period (21-45 days, 3-8 weeks) (slow growing M.0),
growth medium is poured in screw capped bottles (to avoid evaporation of media due to long IP).**
b. Media →(need enriched media because it is fastidious M.O)
 Dorset egg medium. (DEM): enriched media only
 Lowenstein Jensen medium( LJM)*: selective & enriched: contain: 1. egg medium
2. malachit green (selective)
3. cycloheximide (inhibit fungus)
4. lincomycin (antibiotic) →inhibit other Gr+ & Gr - bacteria)
5. glycerol (enhance growth of MTB & inhibit M.bovis)**"or
6. sod. Pyruvate (enhance growth of M.bovis)*
incubate up to7-8 weeks →yellow rough and tough colonies
 Stone brink's media: selective and enriched media.
(Rapid radiometric mycobacterium detection system as BACTEC TB) ****************
5. Blochemical characters:
Nitrate Urease Niacin Na. Glycerol Growth Temp. Growth
(yellow) pyruvate time type
+ + + - + 3-8 37 Eugonic MTB
weeks (heavy G)
- + - + - 3-8 37 Dysgonic M.Bovis
weeks (Scanty)
- - - - 2-6 37-43c Eugonic M.avium
weeks (heavy G)

6. Lab. animal inoculation and pathogenicity tests →

M.avium M.bovis MTB

- + + Guinea pig
- + - Rabbit
- + - Cattle
+ - - Fowl
- + + Man

7. Confirmatory diagnosis →Lymphocytes transformation test, IFN gamma assay, and ELISA or
(PCR)
Vaccination****** →BCG (bacilus Calmette and Gurin) attenuated strain of M.bovis (since 100
year)
Mycobacterium paratuberculosis (Jhones disease)
 signs →emaciation darshoea, sheep.eoat.cattle) Samples are →ilio-cecal valve or rectal swab
or feces →processing of sample →culture on Egg yolk medium that contains mycobactin
protein that stimulates growth)
 (P.M →corrugation of intestine.
 Culture →egg yolk medium + mycobactia***" growth promoter for M. paratubertulosis (incubate
for 18-24 months)
 Field diagnosis allergic test (jhonin test) (I.D) give false +ve up to 75%
Atypical Mycobacteria
Runyon classification
1-acc.to pigment prod. (colonial morphology)
- Schotochromogens (yellow pigment in absence or presence of ight)
- Photochromogens (piement in light only)
- Non chromogens as (M.avium)
Z-acc. To growth rate***
- Rapid grower(less than 7 days) M.fortutum
- Slow grower (over 7 days) + Scoto. As M scrofolacium + photo. As M.marinum
Notes
  Incubation time of T.B is long? Because generation time required for doubling is 20 HOURS
while in E.coli is 20 MINUTES.
 Treatment of T.B (treatment only in human but in animals test and slaughter strategy because
(1. it is Intra-cellular M.O, Z. Chronic disease, 3, Zoonotic transmission) but there is vaccination
by BCG (attenuated strain of M. Bovis).
 T.B resist DECOLOURIZATION by acid and alcohol but not resist acid or alcohol (True)
Niacin production test is very importatt for identification of MTB and depends on treating the
extracts of colonies by means of ANILINE 4% and cyanogen bromide 10% and the presence of
niacin is indicated by appearance of YELLOW color
 Saprophytic mycobacterium often referred as atypical. (T)
1.
McQ queitions
Members of mycobacterla are difficult to be stained by gram due to:
a-lack of cell wall.
b-has waxy coat.
c-unstainable.
d-a and b.
2. Pathogenic Mycobacteria are mainly.
a- acid and alchol fast.
b- alchol fast.
c- non acid, non alchol fast.
3. Atypical mycobacteria differ from typical one in : a-pigmented
b-rapid grower
c-acid and alcohol fast
D-day a and b
4. Culture for pathogenic mycobacteria on L Jensen media still in incubator maximally for.
a-one month at 37c
b-24hr at 37c
c-2 weeks.
d-non of them
5. Mycobacteria are mainly scanty in sputum samples, it must be treated with.
a-bile b- 4% Na hedronide
c-H2so4
6. Tuberculin material contains.
a- tuberculo proteins
b- antituberculous Ab
c-T.B cel wall
d-non of them
Actinomycetales
Actinomycetes & related organisms
• Classification →filamentous actinomycetes (produce true mycelium) = fungus-like bacteria are:
1. MYCOBACTERIUM 2. No cardia 3. Actinomyces 4-stretomycetes
5-corynebacteria equi (Rhodococcus equi)
 All of them except (Actinomyces) has high lipid contents in their cell wall (phospholipid -
sulpholipids - glycolipids) →impermeable to gram stain (but typical as wall of Gr +ve bacteria)
→stained by Zheil nelsen stain (ZN). & resist phagocytosis.
 Actinomyces Nocardia
Anaerobic, long filamentous baccilli Aerobic
ZN -VE (blue bacili & blue background) ZN +VE (pink bacilli & blue background)
-VE On SDA: +VE
Normal flora On Soil
+ Spore: -ve
- Chronic granulomatous mastitis & Abortion
1. Actinomyces bovis = lumpy jaw (Gr +ve) - in cattle Cattle farcy (lymphangitis)
> chronic Pyogranulomatus disease affect Tuberculus like disease (ulcerative lesion)
human and all animals) CHAR. BY
SULPHER GRANULES in the pus
2- Actinemyces pyogenes: summer mastitis

A. pyogene A. bovis character

Some are aerobic( CO2) Anaerobic & Co2 Atomospheric requirment
- - Catalase
- Sulphur yellowish Granules in pus
diphtheroide Filamentous(D) Direct microscopic ex
(+) - CAMP(Chrisite, Atikins and
unch Peterson)
A hazy hemolysis Non hemolytic Hemolysis

Actinobacillosis Actinomycosis
Actinobacillus ligneresi Actynomyces bovis
Woody tongue in cattle Lumpy jaw in cattle
Gr -ve Gr +ve
Not stained by ZN ZN -VE
Aerobic Anaerobic
No sulpher granules CHAR. BY SULPHER GRANULES

Family: Bacillaceae (SPORE forming family

Clostridium Bacillus
Anaerobic (obligatory) Aerobic O2 requirement
Bulging Bulging &Non bulging Spore
- + Catalase

Recent classification: Recently, two groups only (B. cereus group and B. subtilis group) have been
developed according to ability to produce acid from mannitol and their production of lecithinase:
1-B. cereus group unable to produce acid from mannitol and produce lecithinase**
B. cereus group, non bulging by oval spore, Gram +ve Large cells ≥ 1um
(B. cereus group): B. cereus - B. anthracis, -B. mycoides - B. pseudomycoides - B.thuringiensis -
B.weihenstepha-nensis
2-B. subtilis group: bulging, Ellipsoldal Spore, Gr. Variable, Small cells
Produce acid from mannitol and lecithinase negative
The bacillus species have:**(Q):
high medical (emetic toxin; B. cereus).
2- biowarfare (virulence plasmids, B. anthracis),
3- economic (biological insecticides; B. thuringiensis) imponence
Genus: Bacillus

1. Bacillus anthracis (Anthrax bacillus) →sudden death

2. Bacillus anthracoides →(Pseudo Anthrax bacilli)
Anthrax
 Definition →Acute fatal bacteraemia that affects mainly herbivorous animals that cause
(Cutaneous form, Pulmonary form, Severe enteritis)
 Susceptibility →Herbivorous animals (cattle, sheep "except Algerian sheep", horse, camel and
elephant), horse are intermediate, Carnivore & pigs are rare (comparatevily resistant), Birds
are very rare
 Natural pathogencity →
A. Man →Skin (wound or insect bite): Malignant pustule,- Inhalation: wool sorter disease, --
Ingestion: severe enteritis
B. Herbivorous animals →Cattle and sheep (except Algerian sheep) →Sudden death, swollen
spleen, animal die within 1-2 days with no symptoms or after 4 days with acute septicemia,
fever, decrease milk secretion, tarry blood discharge from all natural orifices
-The main routes of spores entry are by ingestion from soil or contaminated foods and by
infection of wounds.
C. Horse →Throat oedema, Oedema at external genetalia
D. Pig →Throat oedema, septicemia
E. Dog, cat →Oedema and severe enteritis
 Resistance →Vegetative form die at 55-65°C while Spore form is very resistant (die at 100 for
10 minutes). **
Toxins →It has 3 toxins (tripartite toxin) : (protective factor & Lethal toxin and Oedema toxin),
Toxin formed only inside host not invitro culture
 Toxin, is a plasmid encoded tripartite protein toxin with protective, lethal and oedema factors
(leukocidal, increas vascular permeability and produce capillary thrombosis)
 The toxin consists of three distinct antigenic components. Each component of the toxin is a
thermolabile protein with a mw of approximately 80kDa (A-B enzymatic-binding structure).
a) Factor I Is the edema factor (EF) which is necessary for the edema producing activity of the
toxin. EF is known to be an inherent (adenylate cyclase)
b) Factor Il is the protective antigen (PA): which has two active (A) domains EF (above) and LE
(below)
c) Factor III is known as the lethal factor (LF) because it is essential for the lethal effects of the
anthrax toxin.
 combinations of two or three of the toxin components yield the following results in experimental
animals Q.
PA+LF combine to produce lethal activity
EF+PA produce edema
EF+LF is inactive
PA+LF+EF produces edema and necrosis and is lethal
What makes B. anthacis fully virulent??? Capsule +3 toxins
 Lab diagnosis →
1. Sampling →Contraindicated to open the carcass, just M.O. find air, it sporulated, resist
dryness and hotness up to 50 years,
Samples as →Blood from tail vein or ear vein or Part of hide or any horney materials
2. Morphological characters
 Gram +ve large bacili (3-8ux1-1.2u), non motile
 Straight rods that arranged in chains →square end (rounded in case of non pathogenic
anthacoides)
 Sporulated → control non bulged spores (formed in culture at 32-34"C under aerobic condition)
(non pathogenic: central - terminal -subterminal bulged and non bulged spore)
 Non motile (all non pathogenic : motile by peritrichous flagella except "B. mucoides"**)
 Capsulated →poly-D-glutamate polypeptide capsule that found in blood, body fluid and media
containing animal protein, serum, 10-25% CO2.
 It can be stained by polychrome M.B →Purplish pink capsule surrounding violet bacili
(Macfadyean' reaction).** (non-pathogenic: non capsulated)
3. Culture characters →***
 Rough form is virulent & smooth form is avirulent
 Cultivation on media that has animal protein (serum) and co2 10-25%
 Grow at 41-43°C, best degree at 37"C
 Grow at slightly alkaline media (7.5-7.8)
 Colonies are 2-3mm in diameter, dull, opaque, rough, with irregular outline that appear as
medusa head or curled hair***
 Fluid medium: pellicle and granular deposite (cotton wool like appearance)
On gelatin medium: inverted fir tree ** (slow gelatin liquefaction (absent in non-pathogenic)
 On blood agar: no hemolysis
 On selective media: Polymyxin lysozyme-EDTA-thallous (PLET) acetate agar or polymixin egg
yolk mannitol bromothymal blue agar (PEMBA) appear as turquoise or peacock blue with egg
yolk precipitation zone (weak).
B.Anthracis show no haemolysis on blood agar, medusa head colonies
4. Biochemical reaction →
 It ferment glucose, maltose, sucrose, trehalose (but not lactose) →acid & no gas
 Salicin sugar fermentation →late and slowly (non-pathogenic : rapid)"
 Starch hydrolysis
 Catalase +ve
 Nitrate +ve
 Litmus milk +ve & and penicillin sensitive**
 Gelatin liquefaction +ve but slow (inverted fir tree after 8 day's)
 H25 -ve
5. Lab animal inoculation →Experimental infection
 Mice, guinea pig, rabbit while rat is resistan →S/C injection resulted in death within 12-30 hours
with signs of septicemia and Dark red or tarry watery uncoagulated blood from all natural
orifices, Spleenomegaly. hepatomegaly, gelatinous oedema at site of injection
 Unsuscetable host & lab, animal for B.anthracis are ...... & rat respectively.
6. Serological tests →
1. Ascoli's test = Ring precipitation test= Thermo precipitation test **
A. Samples are: scraped hide, skin or piece of tissue
B. Applied by: 1- Boiling of sample in 5-10 ml of physiological saline for 15 minutes →Cooling -
> Filtration + Add 1 ml of filtered Ag to equal volume anti-anthrax serum →+ve result is ppt ring
at junction between two fluids
2. Fluorescent Ab technique
• Immunity (vaccination) →
1. Passive Immunity →Hyperimmune serum
2. Active immunity Pasteur 1, Pasteur II, Spore vaccine, Carbozoo vaccine (spore vaccine +
saponin "for dilution - necrosis for limiting spread of infection
3. Simultaneous method

Bacillus Anthracoids
• Nagler's reaction*** (On lactose egg yolk milk agar medium) →B.Cereus and B.mycoides give +ve
result that characterized as pearly opalescence zone surround the colony due to lecithin hydrolysis,
Non lactose fermenter
B.anthracis: weak lecithenase activity-
 Gelatin media: fir tree absent (rapid liquifaction)
• Pathogenicity →Non pathogenic except →B.cereus →food poisoning in man, B.subtilis →severe eye
infection
• N.B →
Sudden death causing M.O b. anthracis (no hemolysis on bl.agar), cl. chaouvii, pasterella hemolytica (no .1
hemolysis on bl.agar)
B. anthracis B. anthracoid
End of bacilli Square Rounded
Spore Central Central / terminal / subterminal

Non bulged Bulged / non bulged

Motility - Motile except mucoides

Capsule + )macfedyean( -

Inverted fir tree on gelatin + -

Salicin sugar fermentation Late & slowly Rapid
Lecithenase activity weak Strong (B. cereus - B. mucoides)
(nagler's)
Penicillin S R