ADDIS ABABA UNIVERSITY COLLEGE OF VETERINARY MEDICINE AND

AGRICULTURE

SUMMARY ON VETERINARY LABORATORY EXPERIENCE PERFORMED DURING THE

EXTERNSHIP PROGRAM OF THE ACADEMIC YEAR 2021/2022

BY

Beranu Girma

Site

Bishoftu college of veterinary medicine and agriculture

SUBMITTED TO AAU-CVMA

APRIL,2022

BISHOFTU,SHEWA,ETHIOPIA

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 Table of Contents
LIST OF TABLES................................................................................................................................. ii
1.INTRODUCTION .............................................................................................................................. 1
2. EXAMINATION OF FECAL SAMPLE .......................................................................................... 2
2.1.Materials ...................................................................................................................................... 2
2.2.Procedure of fecal sample collection ........................................................................................... 2
3. EXAMINATION OF BLOOD SAMPLE ......................................................................................... 3
3.1.materials....................................................................................................................................... 3
3.2.Procedure of blood sample colleection ........................................................................................ 3
4. EXAMINATION OF RUMEN FLUID SAMPLE ............................................................................ 5
4.1.Materials ...................................................................................................................................... 5
4.2.Procedures of rumen fluid collection ........................................................................................... 5
5. EXAMINATION OF URINE SAMPLE ........................................................................................... 7
5.1.Material ........................................................................................................................................ 7
5.2.Procedures of urine sample collection ......................................................................................... 7
6. EXAMINATION OF CLOACAL SWAB SAMPLE ........................................................................ 8
6.1.Materials ...................................................................................................................................... 8
6.2.procedures of cloacal swab collection .......................................................................................... 8
7.MEDIA PREPARATION( NUTRIENT AGAR) ............................................................................... 8
7.1.Material ........................................................................................................................................ 8
7.2.Procedures of media preparation ................................................................................................ 8
7.2.1.Sterilization............................................................................................................................. 8
7.2.2.Preparation............................................................................................................................. 9
8.DIRECT MICROSCOPIC EXAMINATION OF LICE AND TICK ............................................. 11
8.1.Material ...................................................................................................................................... 11
8.2.Collection procedure( for tick) .................................................................................................. 11
8.3.Collection procedures for lice .................................................................................................... 11
9. EXAMINATION OF MILK SAMPLE........................................................................................... 12
9.1.Materials .................................................................................................................................... 12
9.2.Procedures for Collecting Milk Samples ................................................................................... 12
10.CONCLUSION AND RECOMMENDATIONS ............................................................................ 14

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 LIST OF TABLES

Table1 :summary of fecal sample examination…………………………………..……………….2

Table2: summary of blood sample examination.............................................................................4
Table 3:summary of rumen fluid examination…….………………..…….…………………...….5
Table 4:summary of urine sample examination……………...…….…..………...………………..6
Table 5: summary of biochemical test …………..………………………………………………..8
Table6:summary of direct stereomicroscopic examination…..…….………...…………………10
Table7:summary of milk sample examination…………….………..……..…………………….11

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 1.INTRODUCTION

Diagnosis is an art of precisely knowing the cause of a particular disease The diagnosis is based
on accurate history, careful examination of animal, collection of material for laboratory
examination and correlation and interpretation of findings. The Veterinary Laboratory diagnosis
has the great role in diagnosing various disease in animals diagnostic samples which submitted
by veterinarian to serve animal owners and research projects.the veterinary laboratory diagnosis
aids the clinician and veterinarian who work over research and laboratory to confirm diagnosis
Therefore, the most common examination undertaken during practice were
❖ Examination of blood sample
❖ Examination of rumen sample
❖ Examination of feaces sample
❖ Examination of urine sample
❖ Examination milk sample
❖ Cultured colony from cloacal swab examination
❖ Direct stereomicroscopic examination of lice and tick

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 2. EXAMINATION OF FECAL SAMPLE

2.1.Materials

❖ Disposable gloves , marker

2.2.Procedure of fecal sample collection

❖ Restrain the animal

❖ Appropriate disposable gloves should be worn
❖ inserting a moistened finger into the rectum and massaging until the external sphincter
relaxes.
❖ Faecal samples collected from the rectum.
❖ tighten the glove as close to the faeces as possible.
❖ Label andTransport the to the laboratory room
❖ As soon as the faecal samples arrive at the laboratory stored in a refrigerator until they
are processed. If delayed
Table1 :summary of fecal sample examination

N Spp of Lab.technique Purpose Result Interpretation

o animal employed
1 bovine Fecal flotation To identify eggs oval, elongated and pointed strongylus egg
of parasites at both ends egg observed
2 bovine Fecal To identify eggs Ellipsoidal,operculated egg Fasciolla egg
Sedimentation of parasites filled with granular
yellowish brown contents

3 ovine Baermann To confirm dx No larvae was seen the animal more

technique likely free from
lung worm parasite

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 4 bovine Direct fecal To confirm dx No parasite egg was visible The animals would
smear or would not be
free from parasite
because we use
small quantity of
fecal
5 bovine Mc master To determine 8 and 10 strongyle eggs Therefore,the
eggs counting the no. of eggs were counted in the first animals was highly
/gram of feaces and second chamber infested with
respectively. 18x50=900 strongylus spp.
eggs/gram

3. EXAMINATION OF BLOOD SAMPLE

3.1.materials

❖ Disposable gloves,EDTA tubes,vacutainer needles,Clippers,Antiseptic,Gauze,marker

3.2.Procedure of blood sample collection

❖ Restrain animal and Appropriate disposable gloves should be worn

❖ Clip and swipe with antiseptic gauze to remove superficial dirt and debris.
❖ Occlude jugular vein by applying pressure at the base of the jugular groove and visualize
raised vein
❖ With bevel up, insert needle firmly into skin and then to vein at angle
❖ using vacutainer, once needle inserted, stabilize needle and push the vacutainer tube into
hub. If youhave hit the vein, blood will flow freely into tube.
❖ If you have missed the vein, carefully reposition needle, with vacutainer attached, until
vessel penetrated.
❖ Once collection complete, remove vacutainer tube, then, applying pressure over
injection site, remove needle and Dispose of needle in approved container.

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 ❖ apply pressure with gauze in order to ensure adequate haemostasis.
❖ invert the tube several times to mix.
❖ Label and Store to transport
❖ As soon as the blood samples arrive at the laboratory they should be stored in a
refrigerator until they are processed. If delayed
Table2: summary of blood sample examination

No Spp of Lab.techniq Purpose Result Interpretation

animal ue
employed
1 bovine PCV To determine 21% In cattle normal range is
the presence between 24-48%
or absence of The animal was anemic as
anemia result of lower than normal
range
Equine Sahli’s To estimate 14gm/dl The normal 11-14 gm/dl in
2 method hemoglobin equine, so its normal
3 Bovin RBC count Todetermine 5.46x106 RBC/μl lower than the normal
e RBC status value(7x106RBC/μl)
indIcative of anemia
4 bovine WBC count To determine 230 WBCs co unted The result was of normal
WBC status total no.of WBC=230 value(7-12x103WBC/μl of
x50=11.5x103WBC/μl blood)
5 bovine Thin blood To Light blue and deep blue Lymphocytes
smear ( differentiate cytoplasm and nucleus Monocytes
wright stain) WBC respectively
Orange to pink granules

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 4. EXAMINATION OF RUMEN FLUID SAMPLE

4.1.Materials

❖ Mouth gag, Stomach tube, sampling bottle,nose grip,ice back.

4.2.Procedures of rumen fluid collection

❖ Restrain the animal and hold with nose grip. Open the mouth by pulling out the tongue to
one side and by holding the head of animals high
❖ intubate (introduce) orally with aid of mouth gag which have hole centrally, until it
reaches the rumen.
❖ Collect the rumen fluid there will be spontaneous flow of rumen fluid
❖ Label and Store to transport the sample
❖ As soon as the rumen fluid samples arrive at the laboratory stored in a refrigerator until
they are processed. If delayed
Table 3:summary of rumen fluid examination
Spp of animals: bovine

No lab.technique Purpose Result Interpretation

employed
1 dipstick To determine PH value is 8 Normal PH of rumen 6-7 in bovine so
PH of rumen the result higher than normal range
2 Sedimentation To evaluate Sedimented within Normal sedimentation time (4-8 min )
activity test rumen 3 min so rapid sedimentation indicates
microflora inactive microflora
activity
3 Methlyene To determine Decolorize within Normal rumen fluid from cattle
blue reduction anaerobic 15 min decolorized with in 3 minutes leav ing
test fermentative narrow ring of blue color at top layer
metabolism Abnormal reduction time was

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 of bacteria observed (inactive)

4 Cellulose To determine Cotton thread active rumen fluid digest the cellulose
digestion test the ability of dropped within 48 and dropped Within 48-54 hrs. as
microflora to hrs result microflora digest the cellulose
ferment
cellulose
5 Glucose To determine 5ml gas per hr was Normal rate of gas formation 1-2ml
fermentation the ability of formed per hr so higher rate of gas
test microflora to formation(normal fermentative)
ferment
glucose
6 Nitrate To determine Red color Rumen fluid of cattle fed a mixed
reduction test the activity disappear within ration not change color after 5-10 min
of microbe to 10, 20, 30min in in tubeI and 20 min in tube II and 30
that reduce tube 1, tube2 tube min in tube III.as result of this its
nitrate 3 respectively normal fermentive to nitrate
7 Gross To confirm Olive,slight Normally rumen fluid of cattle grossly
examina tion dx viscous ,aromatic Olive, slight ,viscous,aromatic so the
of rumen fluid result was normal
8 Wet mount To determine 10-30 actively There was highly motile and very
Protozoan moving protozoas crowded
activity with different size
were seen. The
ciliates were more
than flagellates.

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 5. EXAMINATION OF URINE SAMPLE

5.1.Material

❖ Disposable gloves,Vacuum container,marker

5.2.Procedures of urine sample collection

❖ Restrain the animal and Label on vacuum container

❖ Place a shallow container under when started to urinate.
❖ As soon as theurine samples arrive at the laboratory they should be stored in a
refrigerator until they are processed. If delayed
Table 4:summary of urine sample examination
Spp of animals:bovine

No Lab.techniqu Purpose Result Interpretation

e employed
1 dipstick To determine urine PH value 8 Normal urine PH value in bovine
PH range 7.5-8.5 so the result higher
than normal
2 Benedict test To determine the orange red Blue-negative
presence or absence color change Green/yellow-trace
of glucose in urine was observed Orange red-moderate
Brickred-large amount of sugar
3 Robert test To determine the no white ring If protein is present there is white
presence or absence formed ring forms at junction of reagent
of protein in urine and urine samplem so the protein
absent
4 USG by refra To determine U SG 1.035 Normally USG ra nge1.025-1.040
ctometer in bovine
5 Gross To evaluate physical Red to brown Normally urine from cattle is

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 examination character of urine cloudy strong transparent and yellow or amber,
of urine odor clear, slight odor of ammonia
red brownish, cloudy,strong odor
indicate abnormal urine

6. EXAMINATION OF CLOACAL SWAB SAMPLE

6.1.Materials

❖ swab,ice back, transporting buffered

6.2.procedures of cloacal swab collection

❖ cloacal swab is taken by inserting a swab into the vent and gently swabbing the mucosal
wall.
❖ The swab should be deeply stained with faecal material.The swab is then placed in
transporting media andstored in iceback

7.MEDIA PREPARATION( NUTRIENT AGAR)

7.1.Material

❖ Conical flask,nutrient agar powder,distilled water,autoclave,boiling water bath,weighting

balance, petridish,,incubator,hot oven,hot plate, spatula,piece of cloth,biosafety
cabinet,water

7.2.Procedures of media preparation

7.2.1.Sterilization

❖ Wash petridish with water and sterilize in hot oven

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 7.2.2.Preparation

❖ Take and suspend 28g of nutrient agar powder in 1000ml of distilled powder(2g powder
for 71.5ml of distilled water)
❖ Mix and dissolve completely by heating on hot plate
❖ Sterilization by Autoclave at 1210c for 15min
❖ After autoclaving put in boiling water bath to equilibrate the temperature to incubator
❖ In biosafety cabinet pour the liquid into petridish and wait until solidify
❖ Incubate at 370c for 24 hrs
❖ After 24hrs inoculate in the biosafety cabinety for culturing of microorganism
❖ After 48 hrs of preparation transfer for purification
Table 5: summary of biochemical test
Species of animals:avian sample taken:cultured colony from cloacal swab

No Lab. Purpose Result Interpretation

technique
employed
1 Catalase To determine the ability of the Copious Positive test indicated by immediate
test( slide organism to produce catalase bubbles bubbles formation
method) enzymes or not produced So ,the bacteria can produce catalase
enzyme
2 Indole test To determine the ability of the Cherry The bacteria express trptophanase
organism to convert to indole red ring enzyme that convert tryptophan to
from tryptophan, formed at indole
top layer
3 MR test To determine the ability of Distinct Red color change indicate organism
bacteria follow mixed acid red color follow mixed acid fermentation
fermentation pathway or not observed pathway
4 VP test To determine if the organism Absence The presence of pink red indicate the
produce or not produce acetyl of pink production of acetyl methyl carbinol
methyl carbinol red color The organism can not produce acetyl

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 methyl carbinol
5 Citrate To determine the ability of changed The organism utilize citrate because
utilization organism utilize citrate as Green- the appearance of blue color indicate
test source of energy blue citrate utilization
6 Catalase To detect ability to produce or Copious Positive test indicated by immediate
test(tube not produce catalase enzyme bubbles appearance of bubbes so the bacteria
method) produced produce catalase enzymes
7 Gram’s To differentiate gram+ve and blue gram +ve appear blue /purple color
staining gram-ve bacteria purple in whereas gram –ve appear red color
color.
8 KOH test To differentiate gram+ve and mixture gram-ve bacteria because thin
gram-ve bacteria become peptidoglycan of gram negative
thick bacteria dissolved by KOH and form
stringy thick, stringy and form long strands
and form
long
strand
9 TSI test To determine the ability to yellow the first indicate the fermentation of
ferment CHO and produce slant and CHO
H2S buttom the second indicate the presence of
Blackeni H2
ng of the
medium
10 Urease To determine the ability to No color Positive test indicated by bright red
test produce urease enzyme or not change color change but now there is no
color change so the organism can
not produce urease enzyme
11 OF test To determine gram negative Both Both open and closed appear yellow
bacteria metabolize glucose open and indicates fermentative i.e the result
either aerocic or fermentative closed Open yellow and closed green
yellow indicates oxidative

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 color
observed
12 Oxidase To determine if the organism Deep The Deep purple blue color indicate
test have cytochrome enzymes or purple the organism containing cytochrome
not blue enzymes
color was
observed

8.DIRECT MICROSCOPIC EXAMINATION OF LICE AND TICK

8.1.Material

❖ Forceps,alcohol ,small jar ,stereomicroscope,petri dish

8.2.Collection procedure( for tick)

❖ restraining the animals,

❖ Using forcep grasp the tick as close to skin as possible.
❖ With the tick firmly grasped, pull it straight upward
❖ After removing the tick, clean the bite area, tweezers, and your hands with rubbing
alcohol.
❖ Place the tick in a jar and keep it.
❖ Soak the tick with alcohol in petri dish and observe under stereomicroscope

8.3.Collection procedures for lice

❖ Restrain the animals and search the lice visual on animal hairwith hand grooming
❖ After collecting the lice placed on petridish and soaked with alcohol observe under
stereomicroscope
Table6:summary of direct stereomicroscopic examination

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 Spp of animals: bovine

No Sample Laboratory Purpose Result Interpretation

taken technique
employed
15 tick Direct To identify spp Long,strong mouthparts. Male Amblyomma
microscopy of tick and its festoons are well developed variegatum
examination sex no adanal shields brightly
ornamented with orange.
16 lice Direct To identify spp Narrow pointed mouthpart Linognathus vituli
microscopic of lice long lateral antennae
examination Black blood-filled,wider
than thorax, lateral spiracles
with brown plates.

9. EXAMINATION OF MILK SAMPLE

9.1.Materials

❖ Sterile vials ,70% alcohol , Cotton ,ice back,Disinfectant ,cloth towels,ink pen

9.2.Procedures for Collecting Milk Samples

❖ Wash your hands and put on new disposable gloves.

❖ Using ink pen label a new sample tube and Keep the sample tube closed until the sample
will be collected.
❖ Make sure that the udder and teats are clean and dry.
❖ Discard 3 to 4 streams of milk
❖ Dip all quarters in an effective premilking teat disinfectant
❖ Dry teats thoroughly with an individual towel.
❖ Scrub teat ends with a cotton soaked in alcohol.

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 ❖ Open the sample tube immediately before the sample is taken. Do not let your
hands or the teat end come into contact with the inside of the tube, including the
lid.
❖ After collecting Immediately put the sample tube in the ice back .
Table7:summary of milk sample examination
Spp of animals: bovine

No Laboratory Purpose Result Interpretation

technique
employed
1 Alcohol test To detect milk There is flake The milk was abnormal due to
abnormality and curd inside presence of flake and curd or
the surface coagulated
2 COB To determine Not coagulation If the milk coagulated Upon heating
the stability of was observed its unstable to heat
milk If the milk not coagulate up on
heating its stable to heat
3 CMT To confirm dx Mixture remain The animal free from mastitis
liquid and no gel
formed

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 10.CONCLUSION AND RECOMMENDATIONS

❖ Veterinary and research laboratories use numerous hazardous materials, including

chemicals and biological agents that present potential hazards to workers, patients, the
public and the environment.
❖ Chemical hazards: toxins, corrosives, flammables, reactives and radioactives
❖ Biological hazards: microbes, animals, plants, and genetically modified agents
Therefore based above conclusion the following recommendation was forwaded
❖ Personnel must wash their hands after handling infectious materials and animals, and
before they leave the laboratory working areas.
❖ Safety glasses, face shields or other protective devices must be worn when is necessary
to protect the eyes and face from splashes, impacting objects and sources of artificial
ultraviolet radiation.
❖ Open-toed foot wear must not be worn in laboratories
❖ Food and/or drink shall not be stored or consumed in laboratories.
❖ The laboratory should be kept neat, clean and free of materials that are not pertinent to
the work.
❖ Work surfaces must be decontaminated after any spill of potentially dangerous material
and at the end of the working day.
❖ All contaminated materials, specimens and cultures must be decontaminated before
disposal or cleaning for reuse.
❖ The laboratory door should be closed when work is in progress and ventilation should be
provided by extracting air from the room (Where biosafety cabinets are used, care shall
be taken to balance ventilation systems.

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