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When we entered the Biosafety and Environment laboratory, we went straight to the incubator and observed the culture media we inoculated yesterday. We observed what we expected that was that the last line of streaking had the thinnest lines and many isolated colonies of bacteria can be observed. These colonies can be further described by using some specific terminologies. First is the form or the basic shape of the colony such as circular, filamentous, etc. The size or the diameter of the colonies can also be measured using a ruler. Some also measure the side view of the colonies or their elevation viewed when the petri dish is turned on one end. On a closer look, the margin or the border of the colony can be magnified and observed as well as for its surface (smooth, glistening, rough, wrinkled or dull). Other colony characteristics that can be observed are its opacity (transparent or clear, opaque, translucent, etc. ) and the pigmentation or color (white, red, purple, etc.).The colonies we observed were similar in overall morphology which means that it is possible that they are of the same group. When colonies are distinct, each colony will represent an individual bacterial cell group.

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We then proceeded to the microbiology room and made stock cultures of the colonies in our culture media. These will be kept so that the microorganisms will be in viable condition or so that there will still be a source of the microorganisms if they will be needed. We observed aseptic techniques such as flaming the loops and then dubbing it on to a bacterial colony. The inoculationg loop or needle is then stabbed on to a small tube containing a culture media and then they will be stored in the refrigerator. Also, some colonies were placed using inoculating needles in small microcentrifuge tubes containing double distilled water. They were then placed in a 95 degrees Celsius water bath and this is the Boiling Method of bacterial DNA extraction.

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While waiting for the microcentrifuge tubes with bacterial colonies, we helped in testing some blood samples sent to the laboratory for Brucella spp. using the RBPT.

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For each of the organ sample’s (lungs, liver, mesenteric lymph node) culture media, 2 representative colonies were taken so there were 6 total samples. After boiling them to extract their DNA, they were then used for PCR mixture preparation. The master mix consists of 56.8 ul of double distilled water, 20 ul of buffer, 40 ul MgCl2, 24 ul dNTP, 8 ul each of forward and reverse primers and 3.2 ul of Taq polymerase. Five microliters of DNA for each sample was then mixed with a corresponding amount of the mixture.

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The PCR tubes containing the PCR mixture were then placed inside the thermal cycler or the PCR machine and the run time was approximately 2 hours. Thermal cyclers are “DNA amplifiers” which regulate the temperature in different cycles (denaturation, annealing and extension). The following steps will not be done yet. Mam Noemi said that the PCR products will still be further processed, will be sequenced through an automated DNA sequencer and then the sequence results will be analysed by the BLAST search engine.

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Immediately after we entered the laboratory, we checked the culture media we incubated yesterday and observe who had the best streaking.

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Mam Noemi then instructed us to make another culture media made from Potato Dextrose Agar (PDA) which is mainly for the culture of fungi. According to its label, it is actually a general purpose medium for molds and yeasts.  It is a recommended medium for plate counts in testing food and dairy products and in growing clinically significant organisms. It has a nutritionally rich base (potato infusion) which encourages growth, pigment production in some dermatophytes and sporulation in molds.

PDA is composed of dehydrated potato infusion and dextrose which encourages the growth of fungi which are usually hard to grow. Agar is the solidifying agent of the medium. We prepared the mixture by adding 500 milliliters of double distilled water to 19.5 grams of commercial PDA powder which contains 20 grams of dextrose, 15 grams of agar and 4 grams of potato starch.

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During our lunch break, we visited one of the 12 regional centers of PCC which are strategically located nationwide, PCC- CLSU. Just like the PCC National Headquarters mission and vision, the center aims to help  improve the state of rural farming through carabao genetic improvement, technology development and dissemination, and establishment of carabao-based enterprises which will help ensure higher income and better nutrition.

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Near PCC-CLSU was another center, the Small Ruminant Center which is headed by a different center director from that of PCC. They also aim to help develop, disseminate and commercialize technologies for the socio-political well-being, technical and economic needs, environmental concerns and cultural demands of the farmers. They continuously develop technologies, produce quality stocks and promote the goat and sheep industry in the country and Luzon in particular.

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When we went back to the laboratory, Mam Noemi instructed us to pour the agar mix we prepared to disposable plastic petri dishes. After we poured it, Mam Noemi closed the biosafety cabinet and turned on the switch for UV light. Mam Noemi said that that was to sterilize microbiological surfaces inside the cabinet as well as the materials and media inside the cabinet. This is an example of an Ultraviolet germicidal irradiation (UVGI) which is a disinfection method which uses short-wavelength ultraviolet(UV-C) light for killing or inactivating microorganisms. UV light can destroy the nucleic acids and thereby disrupting bacterial DNA, leaving them unable to perform vital cellular functions and killing them.

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While waiting for the decontamination, we helped the NVSU interns test the blood samples from the Gene Pool for trypanosomes using a simple microtechnique. The technique uses centrifugation of blood in a capillary tube and observing for trypanosomes in the buffy coat. Other blood protozoa may also be observed through this technique. One of its advantages is that the technique does not alter the morphology of the trypanosomes. The only difficulty we encountered with this technique is that the diffraction of light casts shadows at both sides of the capillary tube and therefore it is difficult to examine the sides. This may reduce the capacity of the technique to detect low numbers of trypanosomes especially if they are at or near the sides.

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We also helped sort out and check all the fecal samples indicated on a list by Doc Chat.

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Mam Noemi acquired the plates she decontaminated in the biosafety cabinet through UV irradiation then we proceeded to the corridor. Mam Noemi said that the occupational health nurse requested in giving them additional health awareness by conducting swab testing to determine the amount of microorganisms present in some equipment (e.g. finger scanner, office door knobs, stair handles and telephones) which many of the employees are using. This simple effort is important since the results will help them create a more suitable preventive action to prevent the spread of germs.

Mam Noemi showed us the materials we need for the activity, some sterile cotton swabs, 9 ml of phosphate buffered solution in sterile conical tubes. We first did Equipment Swabbing by rubbing the swabs back and forth with heavy pressure on some of the equipments in the offices and in the hallway common to all (e.g. office door knobs, finger scanners, telephones, printers, doorbell). We then immediately returned the swabs in dilution fluids and labeling them thereafter.

Next thing we did was to test for the Air Quality to assess if the air within the environment is free from contaminants. We made use of the PDA plates for this test by opening the cover of the culture media and placing the petri dishes at random locations in each room for 15 minutes.

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For our final activity for this day, we helped the NVSU interns process some blood samples for PCR of Surra.

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As we entered the Biosafety and Environment Laboratory, sir Dadz tasked us with the fecalysis of fecal samples from a goat herd. We did the usual routine sedimentation and flotation techniques of fecalysis in the laboratory together with Jakee and Jendy. We observed many Toxocara spp. and Strongyloides spp. ova in one of the fecal samples.

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After our fecalysis, we went on to observe Mam Noemi who was preparing different types of culture media. Culture media are important in microbiological tests since they help grown and obtain pure cultures, count microbial colonies, and cultivate microorganisms selectively. It increases the possibility of achieving accurate and repeatable test results.

These culture media contains gelling agents (agar), nutrients, energy sources, growth promoting factors and other elements such as minerals and salts depending on the type of culture media. There are several categories of culture media sucg as growth media which are designed to grow most bacteria. Other categories include transport media for preserving bacteria during transport and enrichment media which increases the numbers of desired bacteria.

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We helped Mam Noemi in making cultures of microorganisms that we were trying to isolate from organs of deceased water buffaloes submitted to the laboratory for disease diagnosis. The three culture media we used today for that purpose were Mannitol Salt Agar (MSA), MacConkey Agar and Eosin-Methylene Blue (EMB) Agar.

We helped Mam Noemi in making cultures of microorganisms that we were trying to isolate from organs of deceased water buffaloes submitted to the laboratory for disease diagnosis. The three culture media we used today for that purpose were Mannitol Salt Agar (MSA), MacConkey Agar and Eosin-Methylene Blue (EMB) Agar.

                MSA is a selective medium which contains a high concentration of salt (7.5 %) that is inhibitory to most bacteria. It is also a differential medium in that it contains the sugar mannitol and phenol red. Bacteria which are able to ferment mannitol will produce acid turning the red media to yellow (indicative of pathogenic species). MacConkey Agar and EMB are also selective and differential media only differing in components. MacConkey Agar contains bile salts and crystal violet which inhibit the growth of most gram positive organisms. It distinguishes non-lactose fermenters from fermenters due to the presence of lactose and neutral red as the pH indicator. The media also contains gelatin and peptones that prove the essential nutrients for growth. EMB, on the other hand, contains eosin and methylene blue which inhibits gram-positive bacteria and lactose and methylene blue as pH indicator.

Mam Noemi taught us proper streaking techniques. Proper streaking is necessary in isolating a pure strain or species of bacteria. It is ideally a rapid and simple process of dilution isolation. It can be done by using any sterile tool such as a cotton swab but in the laboratory they use inoculation loops and needles. To prevent contamination of the agar plates, we did the streaking inside a biosafety cabinet.

The inoculation loop was first sterilized by placing it inside an incinerator. When the loop is cool, it is dipped in a media containing a piece of the organ sample where many bacterial colonies are present. The inoculation loop was then dragged across the surface of an MSA, MacConkey Agar or EMB Agar back and forth in a zigzag motion until ~30% of the plate was covered. Streaking was done in four directions wherein the heaviest growth should be observed in the first section and the last would have many isolated colonies. The plates were then placed inside the incubator and will be observed the following day.

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For today, we were assigned to Doc Jega at the Reproductive Biotechnology Laboratory and Physiology Unit of PCC. They conduct reproductive technologies there which are helpful tools for the improvement of the herd and also in multiplying desirable characteristics of the herd’s progeny. However, since Doc Jega has still some matters to attend to, we helped NVSU interns Jakee and Jendy first prepare some blood samples submitted at the Biosafety and Environment Laboratory for DNA extraction prior to PCR.

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When Doc Jega arrived, we headed down to the second floor where their laboratory was located. Doc Jega introduced us to BioPRYN^®, a test which offers an alternative and easier way for the confirmation of pregnancy in beef cattle. PCC is testing if this accurate test for pregnancy may be applied to water buffaloes and if they can use it for an early re-breeding program. The test detects for the presence of Pregnancy-Specific Protein B (PSPB) which is a protein in the blood circulation of an animal that is only produced by the placenta containing a growing fetus. In addition, Doc Jega said that it is effective in pregnancy detection as early as 28 days as compared to ultrasonographic detection of pregnancy which is 30 days.

Doc Jega acquired some samples for the test so we can try it. Before starting with the assay, the kit reagents as well as the BioPRYN Assay Plate (PL) were prepared at room temperature. For the assay proper, the first step done was by adding a detector solution using a multi-channel pipette to each well of the PL. Standards were added next using a fresh tip for each solution and were run in duplicate (a Standard Hi and a Standard Lo). The third step was bu adding a sample serum to each of the remaining wells using a fresh tip for each sample. The position of each sample was verified using a printed Plate Grid which goes with the kit. After all those steps were done, the microplate lid was sealed with ParaFilm and was incubated for 2 hours at room temperature (18-24 degrees Celsius). While waiting for those 2 hours, we went out to have lunch.

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After we had our lunch, we decided to visit a nearby center which is the National Freshwater Fisheries Technology Center (NFFTC) of the Bureau of Fisheries and Aquatic Resources (BFAR) located within the Central Luzon State University (CLSU) campus. According to the center’s history, it was established in 1979 as a joint project of BFAR, United States Agency for International Development (USAID) and the Texas A & M University (TAMU). It functions according to the Philippine Fisheries Code of 1998 (RA 8550) to serve as a Philippine Germplasm Reference Collection Center for Tilapia and other freshwater species. It establishes and maintains a collection of a variety of freshwater species in the Philippines as well as from other countries for development of broodstock for future use. Those working at the center also helps in the development and evaluation of different studies on aquaculture production system techniques. 

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When we came back to Doc Jega in the afternoon, we continued the BioPRYN test. The ParaFilm seal was removed and the contents of the wells were removed by striking the inverted plate on a tissue paper (blot drying). The microplates were then loaded in an automatic plate washer machine which washed the plate four times with distilled water. Blot drying was again done after. Then, a second solution was added to the plates which is an enhancer solution. It was added using a multi-channel pipette and then the plate was incubated for 30 minutes after covering it with ParaFilm. Blot drying and washing four times were again done and then a TMB substrate solution was added to each well.  The plate was then incubated for 15 minutes at room temperature. After incubation, a stop solution was added to each well using a multi-channel pipette and the well contents were gently mixed through gently swirling for few a times. The plates were read between 630 and 650 nm using an ELISA plate reader.

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Doc Jega decided to show to us other laboratories of PCC and so we went on laboratory hopping. Our first stop was at the Nutrition Laboratory which is laboratory supporting the research and development activities of PCC. They also provide technical services and information to farmers by the assessment of the nutritive value of feedstuffs and other related samples. They also collaborate with other researchers and even students with regards to new knowledge and information. They help address nutritional problems and deficiencies of ruminants (especially the water buffaloes at the Gene Pool) and other livestock species.

One of the laboratory personnel who were touring us said they perform a test that is familiar to us which is proximate analysis. Components of feed such as dry matter/moisture, ash, crude protein, crude fat and crude fiber were being measured. They do detergent analysis such as acid detergent fiber, neutral detergent fiber and acid detergent lignin and mineral analysis such as Calcium and Phosphorus and other relevant analysis.We met sir Reddik who was getting the measurements of some feedstuff acquired from fistulated water buffaloes at the Gene Pool.

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Beside the Nutrition Laboratory was the Dairy Laboratory which provides services for the analysis of milk components.  It supports the Genetic Improvement Program of PCC. Their services are mainly intended for dairy herds who are participants in the milk recording and performance testing of PCC such as their assisted cooperatives. In contrast to the California Mastitis test which is a subjective test, milk samples are rapidly being analyzed by a milk analyzer for quantitative components such as milk fat percentage, milk protein percentage, lactose, total solids, and somatic cell count (SCC).

According to Mam Paulen, one of the staff working in the laboratory, analysis is usually completed within nine working days from the date of request or submission of milk samples. They can, however, preserve the raw milk samples for up to 7 days at 4-5 degrees Celsius by adding bronopol.Unpreserved samples were immediately placed in a container with ice and analyzed within 2 days or 72 hours after its collection. Farmers were also oriented or trained regarding the proper sampling of milk, handling and storage.

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Shine and I were stationed at the laboratory while our two companions accompanied Tata Mar on the field in doing AI. We immediately processed the fecal sample we collected from the calf yesterday and after the sedimentation technique, we observed multiple Toxocara spp. and Strongyloides spp. ova. Sir Dadz said to coordinate with Doc Cla and do appropriate actions to treat the calf.

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Sir Dadz then tasked us to process the fecal samples we collected from Saranay using the sedimentation and flotation techniques. We asked for the help of the NVSU interns and we did fecalysis all morning. We then prepared to go for lunch and Mam Noemi, Mam Jhena, and Sir Dadz went with us.

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What we were excited to taste was the famous and unique Tilapia-flavored ice cream of CLSU which they sell here. Mam Noemi told us that it won  the Innovation Gold Award during the Salon International de L’Agroalimentaire (SIAL) ASEAN in Manila last 2016. Surprisingly, it has no fishy smell and aftertaste. It was served in a cup and in an ice cream sandwich.

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After taking a break and touring CLSU, we went back to the laboratory and did PCR extraction with the same protocol as we always do.We went home tired but happy as we had more bonding time with our fellow interns and our supervisors.

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We waited at the Biosafety and Environment laboratory for instructions about our activity for today when Sir Dadz told us that Tata Mar and Doc Cla were requesting for two clinicians to accompany them in doing AI. We draw lots among ourselves and Shine and I won and went downstairs immediately because Tata mar and Doc Cla were already ready to go. We went to San Jose City, Nueva Ecija in a farm with several heads of buffaloes. We were astonished to see that not only it is a buffalo farm but also a fish farm particularly tilapia. As I have learned from my previous courses, the more appropriate term for this type of farm is “Integrated Livestock-Fish Farming”. The buffalo pens were constructed on the side of dikes which serve as fish ponds. I can recall from high school that one advantage of such farming system is that manure can be used in hastening the growth of natural food organisms that are being cultured in the pond and thereby increasing food for the fish. Another is that excess livestock feeds may be directly thrown to the pond for utilization of fish. With these advantages, small-scale farmers can lessen the need for buying expensive fish farm inputs or in other words, lowers the cost of producing fish. PCC is near a Tilapia Research Center, and one of their studies concluded that not only does this type of farming produce high yields of tilapia, but also provides an efficient way of disposing livestock manure.

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Doc Cla only asks for hot water whenever they do AI. They pour the hot water in a Thermos cup with an attached thermometer. When the temperature reaches the 38-42 degrees Celsius range, Doc Cla retrieves semen straw from the cryocan and places it in the hot water. She lets it sit for some time and then she cuts the plug, places the straw at the tip of an AI catheter and covers it with straw sheath. She then hands it over to Tata Mar who has already inserted his hand on the rectum to guide the AI catheter up to the body of the uterus where the semen will be deposited. Doc Cla and Tata Mar said that the semen is deposited at the body of the uterus to allow sperm capacitation which is an important factor for a successful conception. As mentioned earlier, PCC technicians conduct AI as one of their banner services and provide it to their sponsored farms or whoever requests for it.

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The next farm we went into was found in the middle of an onion plantation. There were also other vegetable plantations nearby such as tomatoes. This is also another type of integrated farming. The manure from the buffaloes can be used as organic fertilizers to reduce the cost of commercial ones. We did usual activities of pregnancy diagnosis and deworming and administration of vitamins A, D and E. After that, the farmer was very thankful that he gave us some freshly harvested tomatoes and onions.

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At the next farm, we encountered a month-old caracalf with some worms protruding out from its rectum. We took fecal samples with the help of Doc Cla and also adult worms to be examined at the laboratory.

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Most of the adult caracows in the farm showed signs of difficulty in breathing and mucous nasal discharges. Doc Cla suspected enzootic pneumonia through pattern recognition and instructed us to administed tiamulin hydrogen fumarate intramuscularly. Based on the carton label, it is an antibiotic used primarily for swine to treat pneumonia. It is a bacteriostatic antibiotic which becomes bactericidal at higher concentrations. It has good activity against Mycoplasma spp., spirochetes and many gram positive cocci such as Staphylococci spp. and Streptococci spp.

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Aside from rectal palpation, Tata Mar also observes for clear mucus discharge for heat detection and timing of insemination. He just inserts a straw sheath into the vulva and aspirates fluid with the use of a 50-ml syringe attached on the other end.

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Before going back to PCC, we encountered more of the farmer’s buffaloes near a river. Tata Mar instructed that he ties it to a tree and we dewormed and gave them multivitamins.

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When we arrived at the laboratory, we stored our fecal samples at the refrigerator. We also made friends with two interns form Nueva Vizcaya State University, Jakee and Jendy. Mam Noemi then called us outside to share to us some food that Doc Bong left for us. So, we ended our day with a delicious ensaymada!

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Sir Dadz handled us at the laboratory in the morning while Doc Daryl was our handler for the afternoon activity. Sir Dadz tasked us to do fecalysis using both the sedimentation and flotation techniques. We observed both stronglye-type ova and Trichuris spp. ova.

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We went back to LBRAF in the afternoon with Doc Daryl and Sir Chito to collect samples from the newly arrived caracalves yesterday. We were not able to do these activities yesterday as it was already dark when they arrived. As part of the quarantine program, the caracalves have to be checked for parasites using fecalysis, blood parasitism especially Surra, and leptospirosis using urine.  All those samples were collected when the animals were restrained either in a chute or with the help of Kuya Ger.

Fecal samples were obtained by using a rectal glove and inserting the hand intrarectally such as in rectal paplpation. Approximately 10 grams of feces was collected since an extra sample is needed for storage and just 2-3 grams of sample is enough for fecalysis. They were placed in sterile small plastic bags and labelled with the animal’s ID number. Blood samples were collected via the jugular vein. Urine samples were collected in Falcon tubes by waiting for the animals to normally void the urine. All the samples were placed in a chest box with refrigerant prior to transport.

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Doc Daryl was expecting to meet us again today to accompany us at LBRAF but as he has still some things to settle, he allowed us to go the lab to help Mam Noemi in DNA extraction. This step is important prior to PCR as it lyses red blood cells and the nuclei to expose the DNA to be amplified and to remove contaminants.

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We bought lunch on our way there just like yesterday but Sir Chito brought additional ulamfor us. He brought ham and sausage made from carabeef which he said were products processed at PCC in a facility near genepool. It was delicious and tasted like quality export. The difference of carabeef from beef is that it is less fatty and is darker.

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We also encountered a caracow which has aborted its fetus. Due to the abortion, the caracow was constantly straining and Doc Daryl said that maybe the placenta was not yet expelled. The contractions grew stronger and then there was a prolapse of the uterus. Doc Daryl, Sir Chito and Kuya Ger led the team and we assisted them in bringing back the uterus inside. Sir Chito administered local anesthesia which is lidocaine injection at the first intercoccygeal space. We placed the exposed uterus above a clean sack. Doc Daryl instructed us to put cold water and sugar powder to the exposed uterus after washing it to minimize its size for facilitation of bringing it back inside. However, the caracow just made stronger abdominal contractions and the uterus just keeps on going out. Sir Chito then decided to call a buyer and sell the caracow. For the mean time, Sir Chito and Doc Daryl brought the uterus inside the abdominal cavity and sutured the vulva to prevent it from opening again. If the procedure should have been succesful, we would have done uterine flushing with diluted povidone iodine solution which was already prepared by Doc Daryl.

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It was beginning to get dark but we did not yet leave the facility because we were waiting for Tata Naro who was escorting an elf with the newly arrived caracalves for quarantine. One important guideline in transporting animals is that the animals must be restrained with minimal stress and less risk for injuries.

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While the caracalves were being unloaded one by one, Tata Naro and Sir Chito were checking their records. Doc Daryl, on the other hand, was observing them for abnormalities in walking, breathing, etc. One of the farm staff filled the water troughs with clean water and fed the caracalves with silage. Silage is a fodder that was stored for a long time without drying first. Feeding the animals and giving them water help them compensate with the stress of transport and may help them adapt to the new environment.

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We started this week at the Livestock Biotechnology Research and Animal Facility with Doc Daryl and Sir Chito. It is located at San Jose City, Nueva Ecija which is just 30 minutes away from the PCC National Headquarters. We bought lunch on the way there since it is quite far from the highway and from stores. It is quite understandable that it is distant since it served as a quarantine facility for newly arrived caracalves or adult buffaloes. These caracalves are “payments” for the buffalo loan programs of the PCC and can come from any of their cooperatives or sponsored families. Quarantine is a very important aspect of farm management for PCC as it is a preventive measure which ensures that no disease enters the genepool or whatever farm these buffaloes will be loaned to in the future. Quarantine usually lasts for a month or two or more depending on Doc Daryl’s observations, physical exam and laboratory results from the Biosafety and Environment laboratory.

We started our activity which is deworming and blood collection later in the afternoon as the sun was high and it was very hot. That was to ensure that the animals will not have additional stress brought about by the temperature. So before lunch, I just familiarized myself with the facility. It is wide approximately 3 hectares and has at least 5 buildings.  There were buildings for the farm staff, a parking building, building where the pens are located, a building where hay is stored, and a milking parlor. Half of the vacant land is intended for plantations of Napier grass which is harvested by mechanical harvesters and forage cutters and maintained by tillers, tractors and rotavators.

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In the afternoon, started deworming the buffaloes with triclabendazole. We started with the youngest age group and move our way pen-to-pen up to the adults. We used neon pink marker paints so we won’t have to see the ear tags for identification of the dewormed animals. Restraint was quite hard since there is no chute to place the animals in. With the help of KuyaGer and other farm staff, we just lassoed the animals, pushed them on a corner or at the fence and tied them there. The most important points is to prevent the animal’s horns and being careful that the animals may step on you as Sir Chito keeps on reminding us.

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The animals that we tested for Surra using PCR last week which were positive were treated with isometamidium chloride. We also collected blood form the jugular vein and transferred the blood to purple- and red-topped tubes, as we intend to do the Mouse Inoculation Test in case we observe Trypanosoma evansi on the buffy coat after centrifugation of the blood in capillary tubes.                 

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One of our learning last week was that the PCC has a team doing research in finding a pure Philippine Carabao using genotyping and chromosome analysis.

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For today, we went to San Jose City at the National Impact Zone to make the usual extension services such as check-up, pregnancy diagnosis, vitamin administration, deworming and other technical services. The dairy farmers we met are quite familiar since they are the members of the Eastern PMPC, the cooperative where we conducted the last milk test day yesterday. We went with Tata Mar and Doc Cla. Doc Cla was an alumna of UPLB and graduated the year before we entered the college but she was very kind to us as if we have the “LB Vetmed” connection. She taught us a lot of things about the activities by asking us questions and patiently guided us in the skills together with Tata Mar. The first farm where we went to was at Daduyo Dairy Farm. Mr. Daduyo is the chairman of Eastern PMPC. We arrived there at aroung 10 in the morning and first conducted pregnancy diagnosis. Unfortunately, the minimum number of pregnant caracows the chairman was expecting was not met. In a dairy farm setting, there must be a minimum number of caracows to become pregnant to ensure production.

                The caracows in the farm are also fed by-products of corn since they have a corn farm nearby and it is harvest time. The chairman also gave us corn for merienda and pointed us to the next nearby farm.

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At Miguel Dairy Farm, there are a lot of animals which they are rearing. There are ducks, goats, chickens, of course carabao and they also have dogs. Some diseases affect multiple species and transmission is possible. But in the farm, the animals are enclosed in different stalls and the pens are very organized. The owner of the farm said that not only are they the one drinking the milk produce but the goats who were “orphaned” as well.                                                

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The next farm, Sembrano Dairy Farm was far away from the two and luckily we still have our corn in case we get hungry. At this farm, we observed a buffalo whose horns are very long and extending caudoventrally from the head. Tata Mar said that this characteristic is typical for a Brazilian Murrah. Some of the animals also have long hooves which may interfere with walking if not addressed sooner. But, the one we were really curious about are the ones with short, cut tails. Tata Mar said there was an outbreak of Degnala disease a year ago and they had to cut the tails of those affected. The causative agent is a Penicillium fungus which was found in mouldy hay that the farmers feed the animals. It causes gangrenous lesions in the tail so they have to cut it off. Animals affected also present signs like alopecia, lesions on the foot which they first thought of as FMD, and swelling on the ventral region such as the brisket and legs. Other management measures aside from surgery were administration of antibiotics, supplementations and other medications, washing and disinfection of wounds, cleaning of the environment and ensurance of good quality feed.

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Tata Mar wanted us to practice our skills in pregnancy diagnosis and guided us as we conduct it. He also taught us how he acquires measurements inside to predict the age of pregnancy.

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Marcelino’s Farm was our last stop for the day. In this farm, the caracalves have elevated whitish nodular skin lesions on the face and ears which they scratch on posts and fences. Doc Cla just advised to administer ivermectin subcutaneously. There was also a caracalf which is unable to stand and has diarrhea. The owner said that most of his caracalves die if they have diarrhea within 3-5 days of birth. Doc Cla administered multivitamins to the calf, gave some advice to the owner and said to check it back again for follow-up.

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We thought all along that the Thermos® we carry at the back of our vehicle was Tata Mar’s water bottle but it functioned for containing hot water and measured the temperature at which the semen straw will be thawed. Doc Cla picked up a semen straw from the cryocan with liquid nitrogen and placed it in the Thermos® where a thermometer is attached. When it reached the proper temperature, the semen straw was opened, placed in an inseminating tube and then the tube was placed in a sheath. She then handed it over to Tata Mar who was already wearing a rectal glove. Tata Mar did the rectovaginal technique of cervical insemination. With his gloved hand, he located the cervix through the rectal wall. He then inserted the inseminating tube into the vagina and guided it into the cervix by the gloved hand. Doc Cla then gave us a short review of our lessons in Theriogenology and then we headed back to PCC.