• 

• 

Today was sadly our last day of externship. We woke up early to conduct a milk test in a dairy farm in Sitio Lomboy. We went with Doc Chat, Doc Jonathan and Doc Duran and two other vet students from CLSU. It was very early and it was cold when we arrived at the farm. The farm staff already prepared the buckets and the portable milking machines we will be using. Doc Jonathan prepared the 50 ml vials where we will be placing some milk samples, the CMT reagents and the CMT paddles.

• 

• 

The water buffaloes were guided to a double-sided herringbone milking parlor with one side only used for 19 buffaloes at a time. Feeds were placed in the feeders and the buffaloes which were known to be aggressive were tied to the post at the hindlimb. One of the farm staff then started forestripping the teats of each animal after he wiped them with a clean cloth. Labels using tapes were placed on each division to denote each animal for easier identification. We then conducted the CMT while the farm staff acquire milk samples using the paddles. Doc Jonathan was recording our observations.

• 

• 

• 

• 

Lastly, before the animals are led out of the parlor, bulk milk was collected using the portable milking machine.

• 

The teats were dipped on povidone iodine solution after milking to prevent mastitis. The bulk milk was weighed and recorded in kilograms. Bulk milk negative for the CMT was then transferred to a collecting bucket after straining in a cloth and after being sampled in a vial. 

• 

• 

• 

• 

The sun was already shining at us when we finished testing and sampling all the animals.

• 

• 

• 

We went back to the PCC headquarters to be able to submit the milk samples at the Milk and Meat Laboratory for analysis.

• 

• 

• 

• 

• 

• 

In the afternoon, we went back to the same dairy farm with some new vials containing Bronopol, a preservative and an ice box. We arrived at the farm at just the right time for the afternoon milking. We collected negative bulk milk samples, placed them in vials and transported them in an ice box. The milk collected was again for milk analysis.

• 

• 

Today was supposed to be a Milk Test day but the cooperative at Riverside had a power and water shortage and so Doc Chat decided to postpone the testing and so we headed back to the laboratory.

• 

• 

• 

• 

At the laboratory, we helped sir Dadz extract DNA from blood samples and Mam Noemi in doing PCR.

• 

• 

We also observed in making agar gel for electrophoreses.

• 

• 

In the afternoon, we helped Mam Noemi do some subcultures of a bacterial culture and sir Dadz with RBPT.

• 

We started the day by processing fecal samples and observing them under the microscope.

• 

Afterwards, we also extracted DNA from some urine samples.

• 

Some of us were assigned to check the blood samples for microfilaria or Trypanosoma spp. The blood samples were placed in capillary tubes and centrifuged for 5 minutes at 10000 rpm. The buffy coat were observed for the blood parasites under the microscope. The PCV was also acquired and then recorded. The blood samples for those tests were from the purple top tubes containing EDTA.

• 

                The blood samples in the plain vials were used for the RBPT. 25 ul of the serum from those blood samples were mixed with 25 ul of reagent for the Brucella spp. testing.

• 

• 

After observing some coccidia under the microscope during fecalysis, Sir Dadz told us to extract its DNA. We did so by doing the following steps:

1. Add 0.5 ml of fecal sample in a 1.5 MCT.
2. Add glass beads until the 1 ml mark.
3. Add 0.5 ml of lysing solution.
4. Vortex the mixture for 30 seconds and then incubate at 95 degrees Celsius for 15 minutes.
5. Centrifuge at 14 000 rpm for 1 minute. Aspirate the supernatant and transfer into a 1.5 MCT with 500 ul of isopropanol.
6. Centrifuge at 14 000 rpm for 1 minute and then discard the supernatant.
7. Add 500 ul of 70% ethanol and then discard the supernatant.
8. Add 30 ul of DNA rehydration solution.
9. Incubate at 95 degrees Celsius for 10 minutes and then store at 20 degrees Celsius until PCR.

• 

• 

For today, some of the PCC staff is still attending their weekly Monday meeting. So, sir Dadz allowed us to go the National Water Buffalo Gene Pool to meet Doc Jim. We rode our bicycles to the Gene Pool and were met by the biosecurity measures such as the tire bath and the fencing. Doc Jim and the vet students from Central Luzon State University were transferring caracows from pen to another. We also saw Tata Mar and Tata Naro conducting pregnancy diagnosis through rectal palpation just like they do whenever we go out to the National Impact Zone or to other dairy farms.

• 

• 

We went around to see the pens and saw some cattle mixed with the water buffaloes.

• 

• 

It was also feeding time for the caracalves and we saw one albino water buffalo which was a progeny of some albino lineage in the farm.

• 

• 

On our way back to the laboratory, we passed by a small building with pens for goats specifically Boer goats which are meat-type of animals.

• 

Back at the laboratory, we helped Sir Dadz in processing some fecal samples and observed a lot of Fasciola spp. eggs in the samples after sedimentation technique.

• 

• 

Some urine samples also arrived at the laboratory and Sir Dadz instructed us to get the DNA samples. The method was as follows:

1. Transfer 1 ml of urine into a 2 ml tube.
2. Centrifuge 14000 rpm at 10 degrees Celsius for 10 minutes.
3. Aspirate the supernatant.
4. Add 37ul DNA rehydration solution
5. Incubate at 95 degrees Celsius at 10 minutes
6. Keep in -20 degrees Celsius for PCR.