Bactrial isolation

Material *bacteriology media (nutrient agar nutrient broth) *flame *petri dishes * forceps *bacteriological loop * spatula *scissors

*samples from freshly dead bird

Method:1-disscection of bird under aseptic condition 2- collection of samples in sterile dishes 3- making surface sterilization for organs using spatula 4-take loopful from organs 5-cultivate on nutrient broth and incubate at 37 c for 24 h 6- turbidity of broth indicate bacterial growth so , subculture on solid media and incubate in Incubator in inverted position Result uses Media Nutrient agar Macconkey Organism Isolation of different organisms Result Staph , E-coli , strept , salmonella

Isolation of enterobacteraec And differentiate between Lactose fermenter (pink color) Non lactose fermenter (pale color)

Pink color as: E-coli citrobacter enterobacter Pale as : salmonella shigella porteus

Blood agar

Isolation of different organisms : 1-colistridia 2- staph 3- pasteurellosis 4-coryza (H.paragallinarum )

1 double zone of hemolysis (colistridia ) 2-Golden yellow colonies +H2S (staph) 3- dew drop colony +butric acid odour (pasteurellosis) 4-tiny colony along the streaked line of feeder organism(staphyicus ) media by NAD essential for coryza growth

Antibiotic sensitivity test Material *bacteriology media (muller henton specific media - nutrient agar ) *flame *petri dishes Method : 1-disscection of bird under aseptic condition 2- collection of samples in sterile dishes 3- making surface sterilization for organs using spatula 4-take loopful from organs 5-cultivate on nutrient broth and incubate at 37 c for 24 h 6- turbidity of broth indicate bacterial growth so , culture on solid media spreading on all surface of media 7-spread the disk on media and avoid contact in between 8- incubate in incubator in inverted position Result 1-presence of growth inhibition zone + * forceps *bacteriological loop * spatula *scissors

*samples from freshly dead bird

*Antibiotic disk

2- resistance Absent of inhibition zone Uses : For choice the effective drug in case of 1-CRD 2-Salmonella 3-Pastuerelosis

Blood Film

Materials *Microscope *slides *ethyl alcohl *Giemsia stain *samples (Blood)

*Filter paper * cedar oil

Method: 1- put one drop of blood on slide 2- using anthor slide at angle of 45 then spread blood over the slide 3-thining the film by another slide 4-add ethyl alcohol for 5 min for fixation 5-add Giemsia stain for 15 min for staining 6- gentile washing using tap water 7- drying film by filter paper and drop of ceder oil 8- examination the film under microscope Sp CHCH pasteurellosis spirochetosis trypanosoma leukosis Haemoproteus leukocytozoon Plasmodium Inter cellular bi polar bacteria Inter cellular spiral shape Inter cellular spiral flagellated High on of imamature cells Inter cellular caping nucleus Inter cellular micro macro schizont Inter cellular ring form

Impression smear

Materials *Microscope *slides *ethyl alcohl *Giemsia stain *samples (Liver - spleen)

*Filter paper * cedar oil

Methode: 1-cut the organ by Knife 2-move part of organ or squeeze on slide 3-thining the film by another slide 4-add ethyl alcohol for 5 min for fixation 5- add Giemsia stain for 15 min for staining 6- gentie washing using tap water 7- drying film by filter paper and drop of cedar oil 8- examination the film under oil lense Result

Direct fecal smear Material *microscope *slides *cover slides *water

*samples from freshly dead bird (intestine liver )

Method:1-using scissor open affected intestine and exposing the mucosa 2-using cover slide scrap mucosa , take part of intestinal content and put on glass slide 3- put on drop of water on isolation content 4-mixing content with water fix your film by cover slide 5- examine the film low power Sp Ascaridae egg Cappilaria egg Cestode Coccidiosis ChCh Large size with double wall Lemmon shape and operculated Gravid segment contain egg Shizont single wallno space Oocyst double wall with space Smear show both shizont +oocyst middle part of intestine smear show shizont only but oocyst present in ceacum

E- tenella E-necartix

Pullorum test Whole blood agglutination test ( ) Material:*polyvalent stained antigen * porcelain plate Method: 1-on clean porcelain plate put drop of blood and one drop of stained s.pullorum antigen 2-using glass rod make gentle mixing 3-mark reading of result within 30-60sc *whole blood sample *glass rod

Result uses : It is screening filed test for detection of blood carrier bird with s.pullorum gallinarum in Breeder floks at age of 16 w (3-4w) before egg production +ve : appearance of fine granules or sand like within 30-60 sc ------- eradicate -ve : homogenous mixture or formation of sand like after 2 min ----- re test Suspect : formation of sand like after 1 min and before 2 min N.B: Turkey flock use serum (high Ab ) not use whole blood (low Ab ) If more than 10% of flock positive eradicate all flock Flock consider negative after 3 successive negative tests

Rapid serum plate agglutination test (SPA) Material : *Mycopalsma stained antigen * serum sample *glass rod *porcelain plate

Method:1-on clean porcelain plate put one drop of serum and one drop of stained mycoplasma antigen 2-use glass rod make gentle mixing 3- mark reading of result within 2 min

Result uses It's screening filed test used for detection of carrier birds with M. gallisepticum and M.synovae

+ve : appearance of fine granules or sand like within 2 min -------- eradicate -ve : homogenous mixture or formation of sand like after 2 min ------ retest Flock consider negative after 3 successive negative tests False +ve : 1-cross reaction with staph aureus 2- previous vaccination 3- serum complement (heat at 56 c for 15 min ) for destruction

Bursa : Enlargement : leukosis + IBD in early stage Atrophy : Mycotoxin + IBD in late stage

Nerve :
Enlargement : *MD --with loss of striation *vit B2 without loss of striation

GIT :
-Hgric dudenitis - pasteurella -Bloody content in middle part E-necortix Caecum E-tenella -red white spot on serosa Coccidia -Elleptical raised ulcer ND -whitish ulcer with red border clostridium colinum -Necrosis of Intestine clostridium perferinges -Nodules in intestine E-coli MD -proventiculs Hgric= Hgric syndrome ND AI Mycotoxia IBD - Caecal core Coccidia salmonella

Muscle :
Hgric on muscle AI IBD- Mycotoxin CIA -White streaks : Vit E def

Esophagus :
-Cosection : * whitish vit A def if removed No hgric

* yellowish trichomonisis

Longitudinal co section in Duck caused by DVE

Blue Comb Disease = Histomoniosis in turkey = HGr in caecum +Necrosis on liver

Spleen : Mottled spirochetosis Enlarged MD Leukosis

Pancerase: Atrophy Reo Necrosis AI Mouth oral Cavity : Caseated Material Pox vit A Def Ulcer Mycotoxin (T2)

Rabbite : Heamothorax By RVHD in Adult ages Lung Abscess in all ages By RBHD

Dr.Sherif