Large Ruminant Practice at USM-

Philippine Carabao Center

Kabacan, Cotabato
February 03-21 2020

Bandoy, Shane Joy U.

Toraja, Keneth Jay O.
Tribo, Jeza M.
 PHILIPPINE CARABAO CENTER ( PCC)

Philippine Carabao Center (also known as Sentro ng Kalabaw sa Pilipinas) was

established by the virtue of the Republic Act No. 7307 better known as the “Philippine
Carabao Act of 1992” and it is reinforced by the Department of Agriculture. The institution
aims to develop and improve the carabao industry in the Philippines through selection,
genetic improvement and breeding resulting to the distribution of improved carabaos
(water buffaloes to farmers, thus, the improvement of rural farming.
Philippines Carabao Center – University of Southern Mindanao headed by Dir.
Benjamin John C. Basilio is one of the 12 operating centers of PCC all over the
Philippines. The institution covers 35.47 hectares of land, equipped with infrastructure
and facilities for dairy processing and monitoring and maintenance.

Mission
Improving the general well-being of rural farming communities through carabao
genetic improvement, technology development, and dissemination and establishment of
carabao based enterprises, thus, ensuring higher income and better nutrition.

Vision
A premier research and development institution profitable and sustainable
carabao-based enterprises designed to improve the income and nutrition of rural farming
communities.
Summary of Activities
The following are the activities performed by the interns throughout the duration of
the training:

Date Activities
January 30, 2020 - Body measuring and weighing of the
animals
February 3, 2020 - Orientation
- Pre examination
- PCC Tour
- Milking and Milk collection
observation
- AI observation
- Administration of Dinoprost
(Lutalyse) for estrus synchronization
February 4, 2020 - Deworming
- Administration of anti-Surra drug
February 5, 2020 - Milk Test
- Ear tagging
February 6, 2020 - Caudal Fold TB test
- Attended Seminar (Artificial
Insemination Forum)
February 9, 2020 - TB Test result reading
February 10, 2020 - Blood Collection
- Blood Examination
February 11, 2020 - Milk Sampling
February 12, 2020 - Ear Notching
February 13, 2020 - Nose Roping
February 14, 2020 - Caudal Fold TB test at Pres. Roxas
- Blood Collection and Examination
February 17, 2020 - Necropsy
- Application of Fly Control trap
- TB test result reading
February 18, 2020 -
February 19, 2020 - TB test and Fecal collection
February 20, 2020 - Fecalysis
February 21, 2020 -
Activities
a. Body measuring and weighing of the animals

The interns, with the help of the personnels, put the animals inside the chute.
Then the animals were weighed and the heart girth was measured, body length and
the height of the buffaloes were also taken.

Fig 1. Weighing scale Fig 2. Weighing and body

measuring the animal

b. Milking and milk collection observation

Milking - is the act of removing milk from the mammary glands of dairy
buffaloes. In USM-PCC’s Institutional herd, collection of milk was performed twice
daily (in the morning and in the afternoon) with an average of 40 liters of milk.
Procedure:

 Before milking, the buffaloes were bathed and cleaned thoroughly to

reduce presence of bacterial contaminants.

 The teats are wiped with clean cloth and were massage gently to stimulate
milk letdown. Then, the teat cups were attached to the buffaloes teat.

 The cups alternate between vacuum and normal air pressure to extract the
milk.

 After milk collection, the teats were dipped in povidone iodine to prevent
infection.
 Fig 3. Milk Collection Fig 4. Milk collection (hand
(milking machine) milking)

c. Estrus Synchronization - hormones are commonly used to manipulate the

estrus cycle.

Procedure:
 The animals were restrained properly. The technician palpates and check
if the animal is pregnant or ready and fit for estrus synchronization for the
administration of Dinoprost Tromethamine( Lutalyse).
 If the animal was fit for estrus synchronization, 5ml of lutalyse was given
intramusculary.

Fig 5. Photo of the drug Fig 6. Drug Administration

(Lutalyse)
d. AI observation
AI is a process by which sperms are collected from the male, processed,
stored and artificially introduced into the females reproductive tract for the
purpose of conception.

Procedure:
 The animal was restrained properly for easy access to the rectum and
reproductive tract.
 Thawed semen was loaded into the AI gun
 The AI gun was inserted into the vagina with one hand guiding the AI gun
from the rectum
 Locate the body of the uterus and load the semen.

Fig 7. Palpation of ovaries Fig 8. AI demonstration

e. Deworming
It is also known as worming, drenching or dehelmintization. It is the act of
giving an anthelminthic drug to the animal to get rid of helminth parasites such as
roundworms, flukes and tapeworm. Deworming is done regularly at PCC (once a
month).
Procedure:

 The animals were restrained properly for easier administration of drugs.

 Using anthelmintic Albendazole with 15% pampoo, the buffaloes were

deworm orally, 1ml/5kg BW.
 Fig 9. Photo of the drug Fig 10. Drug administration
(Albendazole)

f. Administration of Anti-Surra drug

Surra is transmitted by other biting flies that are found within and outside
tsetse fly areas.

Procedure:
 Restrain the animal properly prior to the administration of the drug.

 Using the drug trypamidium-samorin to cure and prevent trypanosomiasis,

the drug were given to the buffaloes intramuscularly , 1ml/20kg BW.

Fig 9. Photo of the drug Fig 10. Drug administration

(Trypamodium-Samorin)
g. Quality Milk testing
Is an important component of any milk processing industry whether small,
medium and large scale. The PCC milk test procedure includes:

Fig 11. Photos while performing

the milk test procedure

1. Organoleptic
 Color - Observe the color of the milk in the milk container. Buffaloes
milk is white in color. Reject milk with color other than white.
 Odor- Smell the milk. Milk of good quality has clean, milky smell.
Reject milk with objectionable odor such as smell of corn, soap,
acid, rancid, etc.

2. Alcohol Precipitation Test (APT)

 Put 1ml of alcohol into the test tube
 Add the same amount of milk into the test tube
 Mix the two liquids and check for any clot.

Interpretation of the result:

 Positive- develop milk clots; may have increased acidity, mineral

imbalance (colostrum) or lactational disorder
 Negative- no change in the appearance of milk, partial evidence of heat
stability in milk.

3. pH (Acidity)
 Standardize pH meter with buffer solution( pH 7.0 and pH 4.0)
 Pour a small volume of milk into a beaker
 Dip the electrode of the pH meter
 Wait for the reading to stop
 Record pH display
  Wash electrode with distilled water before dipping it into the second
sample.

Interpretation of the result:

 Milk of good quality has an average pH range of 6.6 to 6.8
 Milk with pH lesser than 6.4 is considered acidic or may contain
colostrum milk. If COB – and passed organoleptic test, the milk can still
be processed into pastillas de leche.
 Milk with pH greater than 7.0 is considered basic and suspected to be
mastitic milk. If COB- and passed organoleptic test, the milk can still be
process into pastillas de leche.

4. Sediment test (centrifuge) - test for the presence of contaminants in milk such
as soil, dirt, manure and blood.

Procedure:
 Place 15ml of milk sample in a glass test tube. Cover tightly with
screw cap
 Place the tube in a centrifuge( in an opposite direction)
 Set the centrifuge to atleast 3000rpm and turn on for 5 minutes.
 Turn off after 5 minutes.
 Observe for presence of contaminants which settles at a bottom of
the tubes.
Interpretation of the results:
 Light to dark brown/ black sediments indicates presence of soil
 Reddish sediment indicates presence of blood
 Yellow sediments indicates presence of pus
 Reject milk with dark brown, Black, yellow and red sediment.

h. Ear tagging - Ear tag is a plastic or metal object used for identification of
domestic livestock and other animals.

Procedure:

 The animal was properly restrained to avoid harm and stress. Also
done to perform the procedure easier.
 The ear was grasped properly. Large vessels were avoided to
reduce bleeding.
 Ear tag plier was used to place the labeled tags on the ear of the
animal.
 After performing the procedure the area was disinfected with
povidone iodine.
 Fig 12. Ear tagging

i. Caudal fold TB test- it is the primary screening used to identify buffalo infected
with TB.
Procedure:
 The animal was restrained properly. Then, the tail was tightly hold
upwards.
 The caudal fold area to be injected was properly disinfected.
 It measures the immune response to Mycobacterium bovis using
intradermal injection of 0.1 ml Purified Protein Derivative (PPD) of
tuberculin into the skin of the caudal fold (the fold of skin at the base of
the tail). After administration, the injection site was measured using
vernier caliper.
 After 72 hours, the injection site were inspected and palpated. Any
swelling at the injection site was measured using a vernier caliper. An
increased in size (>4 mm) from the previous measurement was
classified as a responder.
 Fig 13. Photos of the drug and drug
administration

Table 1. Caudal Fold test result in PCC buffaloes

No. Animal ID Skin fold thickness Difference in skin

fold thickness
Before After
(mm)
testing(mm) testing(mm)
1 2UM1500 12 12 0
2 2UM7005 11 12 1
3 2UM15008 10 12 2
4 2UM07024 11 16 5
5 2UM13027 11 12 1
6 2UM13043 11 11 0
7 5UMC105 13 14 1
8 5UMC107 11 13 2
9 2UM16009 13 18 5
10 11238 12 14 2
11 64052 8 8 0
12 2UM11033 11 12 1
13 14047 11 12 1
14 2UM08001 13 13 0
15 5UMC106 9 11 2
16 5UMC109 13 13 0
17 14043 15 15 0
18 11037 7 8 1
19 07007 8 14 6
 20 11306 10 11 1
21 13004 12 14 2
22 10022 12 12 0
23 5UMC191005 12 13 1
24 5UMC108 11 12 1

Table 2. Caudal test result in Sta. Catalina buffaloes

No. Animal ID Skin fold thickness Difference in

skin fold
Before After thickness
testing(mm) testing(mm) (mm)
1 5UMC19103 11 12 1
2 5UMC19095 9 10 1
3 5UMC19096 8 9 1
4 5UMC19099 10 11 1
5 5UMC19098 9 9 0
6 5UMC19094 9 10 1
7 5UMC19104 11 14 3
8 2UMC16043 1O 11 1
9 5UMC19097 10 11 1

j. Seminar (Artificial Insemination Forum)

The seminar aims to increase AI efficiency in Cattle and buffaloes. Major
issues including factors affecting Artificial breeding:
Female animal - reproductive anatomy and physiology
- Genetics and species
Management- nutrition and body score
- Heat stress
- Lactation stage
- Semen handling and quality
- heat detection and timing
- technical expertise
- estrus synchronization method
- animal health
- post-breeding management
- replacement animals ( calves)

Fig 14. Photos during the seminar

k. Blood Collection and Examination (Blood Parasites)
Blood samples were collected via the jugular vein of the buffalo. The collected
blood samples were placed on the vacutainer tubes with EDTA (purple top). The
samples were then put inside an icebox. After collection, blood samples were
subjected to blood smear and microhematocrit centrifugation technique (MHCT). After
the following preparations, these samples were examined under the microscope and
were examined for the presences of blood parasites.
Materials used:
Vacutainer tube (purple top), Syringe, Cotton and Alcohol

Procedure:

 Restrain the animal properly prior to collection of the blood to avoid stress
and for safe blood collection.
 Using multi sample needle with adapter blood was withdrawn from the
jugular vein of the buffalo.
 Collected bloods were transferred to purple top vacutainers.
Fig 15. Blood collection
 i. Blood smear
Procedure:

 Place a drop of blood into a clean slide

 With your hand placed the smooth clean edge of a spreader slide on
the specimen slide, just in front of the blood drop.

 Hold a spreader slide at a 30 0 and 450 angle and draw it back against
the drop of blood.

 Allow the blood to spread almost to the edges of the slide.

 Push the spread forward with one light, smooth moderate speed.

 Label one edge with patient name and date.

ii. Giemsa stain
Procedure

 Filter the stain before use. Make a 1:10 dilution of the stain. Giemsa
stain 1 ml buffer for Giemsa stain 9ml.

 Mix thoroughly

 Fix the smear with Methanol AR for 5 minutes. Air dry.

 Flood the smear with staining solution for 20 minutes.

 Wash gently with tap water. Allow these slides to dry

 Examine the slides under oil immersion objective.

iii. Microhematocrit Centrifugation Technique( MHCT) for Surra in Buffaloes
Blood samples were obtained from buffaloes in the PCC on February
10, 2020 (six samples) and on February 14, 2020 at Sta. Catalina, Pres.
Roxas, Cotabato (nine samples). MHCT is commonly used technique for a
routine inspection of trypanosomes in blood usually in a buffy coat after
centrifugation.
Procedure:

 Collect the blood into capillary tube

 Seal at the dry end and centrifuge with the seal end down for 5
minutes.
 Using a breaker the capillary tube was divided on the area where the
buffy coat is located.
 Examine under the microscope(x100-400).
 Record your results.
Table 3.Blood parasites examination data on February 10, 2020

No. Blood Smear MHCT

1 - -
2 - -
3 - -
4 - -
5 - -
6 - -

Table 3.Blood parasites examination data on February 14, 2020
No. Animal PCV Blood Smear MHCT
ID/Sample ID
1 19094 30 - -
2 19104 33 - -
3 19096 40 - -
4 19098 33 - -
5 19097 30 - -
6 19099 32 - -
7 19103 31 - -
8 19095 29 - -
9 16045 37 - -



l. Milk Sampling- milk samples were transferred from one container to another for seven
times to evenly distributes milk components. It was done to check the fat, protein and
somatic cell count. Sample were transfer to a tube and for preservation of the milk
samples, 1 piece of bronopol was added. To avoid spilling and entry of the water, a
parafilm was used to seal the tube and was put on an ice box.

Fig 16. Photo of materials used Fig 17. Milk sampling

and Bronopol
m. Ear notching- is a method of identification of a buffalo in which they are identify based
on their birth order within a given farrowing. . It involves removing V-shaped or U-shaped
portions of the ear rim in a specific and individual combination of positions using a special
ear notching pair of pliers.

Fig 18. Photos while performing ear notching

Materials used:

Disinfectant (alcohol), cotton, ear notching pliers, betadine and wound spray

Procedure:

 The ear notching chart was familiarized to perform the correct position and
combination of notches before doing the procedure
 The calves were restrained properly to reduce stress and to perform the
procedure easily.
 Disinfect the ear prior to the procedure.
 Using the ear notching pliers the clipped must be position properly.
 To minimize bleeding the blood vessels on the pinna of the ear were avoided.
 Wound was disinfected using povidone iodine then applies also wound spray
after doing such procedure to avoid infection and allow fast healing.

n. Nose Roping- Involves piercing an animal’s nasal septum in front of the cartilage using
a nose tong. The purpose of this procedure is to easily control an animal by a person
alone.
 Fig 19. Nose roping

Procedure:
 Restrain the animal properly before performing the procedure.
 Locate the nasal septum cartilage using hand.
 Using a nose tong the cartilage was pierced.
 A rope was passed through the hole and was tied by a knot.
 The rope was permanently fastened behind the animals head below the
base of the horns.

o. Necropsy - a surgical examination of animal body to identify the cause of death of the
animal.

p. Fecalysis - for identifying animals with intestinal parasites, fecalysis was routinely done
by the USM PCC. After treatment, fecalysis were done to monitor and evaluate the efficacy
of the drug used.
A total of 12 fecal samples were collected from 12 different carabaos through the rectum.
Fecal samples were brought to the College of Veterinary Medicine - Parasitology Laboratory for
fecalysis. The samples were examined using Salt flotation and Macmaster egg counting
technique. Below are the tests and procedures performed in examining the samples.

Flotation Technique

Flotation Technique is a qualitative test for the detection of nematode and

cestode eggs. This is a useful method to use in preliminary surveys to establish which
parasite groups are present.

Procedure:

 About 2 grams of feces was weighed using digital weighing scale and 2.5ml of
saline solution was added.
  Feces were broken up with a tongue depressor. About 27.5 ml of saline solution
was added and was thoroughly mixed.
 Then, it was sieve for 15 times. Fecal suspensions were poured into test tube
until formation of meniscus is noticeable.
 Cover slip was carefully placed on the top of the tube and was left for 5 minutes.
The cover slip was carefully lifted off the test tube together with the drop of fluid
adhering to it and was placed on a clean glass slide.
 Prepared samples were examined under the compound microscope at 10 x 10
magnification.

McMaster Counting Technique

McMaster counting technique is a quantitative technique to determine the

number of eggs present per gram of feces (EPG).

Procedure:

 Weigh 2 grams of feces and place into a small container.

 Add 2.5 ml of tap water and use a tongue depressor to mash and softened the
feces.
 Add 47.5 mL of saturated sodium chloride in a container mixed it thoroughly
with the fecal sample.
 Sieve the fecal suspension through a strainer 10 – 15 times.
 Used a pipette to fill the McMaster Counting Chamber with the fecal sample.
 Wait for 5 minutes in order for the eggs to float.
 Examine the sample under the microscope at low power magnification of the
objective lens.
Table 4. Fecalysis results
No. ID Number Flotation Technique McMaster Technique
Egg Per Severity of
Gram Infection
( EPG)
1 18008 - -
2 19004 - -
3 18003 -
4 18007 Strongyle egg 50 Light
5 19018 - -
6 17021 - -
7 15005 - -
8 17005 - -
9 19004 - -
10 18005 - -
11 16018 - -
12 19039 - -

Fig 20. Photo while doing Fig 21. Photo of strongyle

fecalysis eggs under the microscope

Assessment of OJT/Practicum Program

New Knowledge/Attitude and Skills Acquired

 Improved skill in blood collection through jugular vein

 Learning how to measure and weighing the animals correctly and properly
 Observing the Artificial insemination
 Skills in blood collection, techniques for drug administration, ear notching, ear
tagging, nose roping and restraining was improved
 Learned how to do milk test and milk sampling.
 Able to learn how to perform TB test using the caudal fold test

Theories/Practice actually seen during OJT

 The actual milking process was also observed during the internship
period.
 Milking of the cows were much easier and faster using milking machines
than hand milking
 Interns seen how to do AI and estrus synchronization
 Interns also observed the administration of drugs and dewormers while
the animals were not restrained using a chute.

Benefits Gained

 PCC at USM is close to the boarding house of the student which

decreases the expenses of the interns. Interns acquired additional
knowledge and skills in blood collection, ear tagging, ear notching, nose
roping and administration of drug through hands on experience

Problem Encountered

 Difficulty in restraining buffaloes during blood collection

Recommendations

 Training and additional techniques for proper restraining of buffaloes must

be taught to the interns.