POSTMORTAL DIAGNOSTIC IN EXOTIC BIRDS Prof. Dr. dr hc Gerry M. Dorrestein Dutch Research Institute for Birds and Exotic Animals Veldhoven, The Netherlands Internacionalizace výuky veterinární medicíny jako cesta na evropský trh práce projekt è. CZ.1.07/2.2.00/28.0288
Introduction NOIVBD private pathology laboratory 1200 cases each year 79% avian species 58% necropsies Rest formalin material, faecal samples, etc NOIVBD molecular laboratory Only Avian Bornavirus Diagnostics PCR and elisa (p40) Epidemiological studies www.noivbd.nl
Statistics all mammals birds reptiles amphibians fish 5581 total necropsy 772 14% 22% 4406 79% 58% 186 3% 16% 13 0% 46% 204 4% 13% www.noivbd.nl
Between the different classes there is different anatomy
Introduction Not all patients will survive! Therefore we do a post mortem exam The key of a PME is to find an explanation for the clinical findings And vice versa: the clinical finding indicates pathological changes
A squirrel showing this symptom
Why post-mortem examination? To find out the cause of death To diagnose the problem in a collection As sentinel animals in a collection To confirm the clinical diagnosis To evaluate/understand therapeutic failure To prevent spreading of an infectious disease (other animals or people) New emerging diseases For anatomical-clinical study or curiosity
Why do animals die? Four basic essential needs Why do we live, technically?? What are the basic requirements for cell function?? ZooAnimal Pathology 8/69
Basic requirements for life 1. Oxygen 2. Energy 3. Water Results in membrane function-stability 4. Toxins attacking membranes ZooAnimal Pathology 9/69
Basic requirements for life Therefore Evaluate your findings related to these basic requirements ZooAnimal Pathology10/69
Why post-mortem examination? But the main issue is to understand what is going on!!! Understand what you see. Know what to expect based on clinical information
What do we have before we start the PME? History Signs and symptoms Imaging information Laboratory results Treatment results >>>> = INFORMATION
In addition we have knowledge = our veterinary education, pathology training, etc This should trigger our medical and pathophysiological thinking!!! Resulting in: Information Knowledge hypothesis Imagination But do NOT jump to CONCLUSIONS!
The solution To understand the problem in the animal Use educated imagination/phantasy
What do we need before starting a PME? As much history/information as possible Information about housing and feeding New animal introductions?? All clinical data and treatments What does the owner think that happened? Think of a DDx and collect the right material for confirming the different possibilities Have the animal/material fresh ASAP
How are we supposed to do it? Analyze the history (use books/internet) Find and explain (understand) changes at PME Formulate a theory!! (hypothesis or lie. not a diagnosis) I-do-not-know is not accepted Find the truth behind the lie (hypothesis) Use direct tests (in the necropsy room) Use bacteriology, histology, etc to confirm Formulate keywords (for google, pubmed, etc)
Example (Tamias sibericus) Hypothesis?
Use direct techniques Cytology smear Wet mount
Auxillary techniques Confirmation with histology Diagnose: Capillaria hepatica
Example: Hoesemys spinosa Hypothesis?
Use direct techiques! lung Wet mount organ Stained smears gut What is your hypothesis? How to continue?
Culture on sab Geotrichum sp
Post-mortem examination
Basic lay-out for a necropsy of a small animal 1. Sharp scalpels 2. Forceps and scissors 3. Bone cutters 4. Formalin 5. Alcohol 6. Saline 7. Containers 8. Glass slides 9. Burner
Basic lay-out for a necropsy of a small animal 1. Culture media 2. Oses 3. Cleaning fluid 4. Small plates 5. Marker 6. Stickers 7. Ruler 8. Checklist 9. Paper towels 10.The animal
Collect for additional tests
Cytology as Instant Diagnostic Tool Rapid, inexpensive method Easily done in any practical setting But it needs some practicing Microscopic information about a process Often an etiologic agent can be found Some diagnosis cannot be made without Does not replace histology
Preparation of the slides
Staining of the slide Hemacolor Dry at the air sol 1: 10-60 sec fixation sol 2: 10 sec till orange sol 3: 10-30 sec till purple-blue sol 4: 4-5 sec buffer (ph 7) no rinsing with water in between
View with microscope Condensor fully open screen with obj 10x select thin spot search with 100x immersion oil-lens
Standard selection Clinic blood ascitis fluid cysts contents abcess material conjunctivitis dermatitis etc, etc, etc Necropsy liver spleen lung rectum any altered organ or lesion
Giardia budgerigar
Atoxoplasma (I. serini)
Specific staining Stamp, Ziehl Neelson
Laboratory diagnostics PROVIDES QUICK AND USEFUL INFORMATION BUT BE SURE YOU ARE TAKING THE RIGHT SAMPLE AND DOUBLE CHECK THE RESULTS
Laboratory testing Faeces (crop,stomach,intestine), urine, se- & excreta Macroscopy Microscopy Abnormal tissue Colour, consistency, aspect, smell Wet mount (NaCl 0.9%, amphibians 0.6%) Enrichment (coccidia, worm-eggs) Stained smears, cytology Cytology Histology Electron microscopy (EM) Microbiology, serology, molecular biology (PCR)
Basic microbiology but first cytology!! Aerobic culture Bloodagar Selective agar (Gram -) Serumbroth Enrichment fluid (salmonella) Media for fungi Incubator 37C
Additional determination Proteus sp.
Dermatitis Green Iguana (PA 09 0860) Culture: Pure Klebsiella pneumonia
Chrysosporium anamorph of Nannizziopsis vriesii (CANV)
PA 07 0454 Ara maracana Diagnose? Next??
Illustration of false culture results
Result Bacteriology Laboratory E. coli Sensitive to Not sensitive to Amoxy Clav Cefquinome, Cephalexine Cefoperasone Colistine, +/- Doxy Florfenicol, Furazolidone Lincomycine, Neomycine Tulathromycine, Ceftiofur Ampi, Amoxy Enroflox, Flumequine Tetracycline TMPS Tylosine Difloxacin
However this is the cytology!
Result Bacteriology Laboratory E. coli Sensitive to Not sensitive to Amoxy Clav Cefquinome, Cephalexine Cefoperasone Colistine, +/- Doxy Florfenicol, Furazolidone Lincomycine, Neomycine Tulathromycine, Ceftiofur Ampi, Amoxy Enroflox, Flumequine Tetracycline TMPS Tylosine Difloxacin
Use a dissecting microscope
Use a dissecting microscope YF amazon nephritis and gout
Neophema with kidney problems
Neophema with kidney problems
Neophema with kidney problems
Neophema with kidney problems Severe interstitial nephritis with uric acid deposits
Articular gout Joints leg, thick, swollen Red, warm painful
a) Tuberculosis Some different livers b) Leucosis c) Amyloidosis d) Lipidosis
Some different livers a) Fibrosis b) Liver necrosis c) Circovirus d) Psittacosis
What do I collect and how? Standard collection for histopathology (see protocol) in buffered formalin 4% Cytology: liver, spleen, lungs, rectum plus anything you see abnormal or interesting Microbiology: based on findings and cytology Freezer (-20/-70C) for virology, toxicology Alcohol 70% (GTC buffer RNA virus) for PCR or on advise of the laboratory Parasitology in alcohol 70% plus 10% glycerine
Additional sampling at necropsy Fixation of tissues in formalin 4% Tissue suspension in alc 70%/GTC Liver, spleen, kidney, etc -20% Specific sampling related to DDX Sampling related to research Sampling related to stud-book Sampling for eductional projects Histology PCR Virus, tox
Summary basis diagnostic tools Necropsy Wet mounts Stained impressions www.noivbd.nl
Necropsy www.noivbd.nl
Diagnostic tools www.noivbd.nl
Wet mounts www.noivbd.nl
Stained impressions www.noivbd.nl
Use of stereo microscope
Know your anatomy e.g. pigeon crop milk www.noivbd.nl
Exercise: A Gouldian finch www.noivbd.nl
After removing first layer www.noivbd.nl
Problem area = stomach www.noivbd.nl
Proventricular contents yeasts and cocci www.noivbd.nl
Other problem area = duodenum www.noivbd.nl
In duodenum no bacteria Starch www.noivbd.nl
Problem area = small intestines www.noivbd.nl
Contents intestines yeasts and mixed bacteria Campylobacter sp www.noivbd.nl
Final conclusions Male GF in poor condition Main problem area GI tract Bacterial and yeast contamination stomach Undigested starch in the duodenum Undigested starch and bacteria in small intestine >> starvation Including Campylobacter sp. Diagnosis: Campyobacteriosis www.noivbd.nl
In conclusion Why is the PME done? Always use a protocol or checklist Work with as fresh material as possible See abnormalities and make pictures Make wet mounts and smears Fix tissues in adequate amount of formalin (size max.5 cm) Formulate a hypothesis and make a report www.noivbd.nl
Questions? www.noivbd.nl