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Today was different since it is the first time we went to a cooperative together as four. We headed to Barangay Sibut, San Jose City, Nueva Ecija in one of the cooperatives there, Eastern Primary Multi-Purpose Cooperative. PCC is highly supportive of the establishment of organizations in local communities such as cooperatives. These cooperatives are envisioned to become independent and effective avenues in the promotion of enterprises for the carabao industry. Smallholder farmers or families can find ways through these cooperatives for creating business which is usually only for commercial producers.These farmers consolidate,grade and process (sometimes even trade) water buffalo farm products such as meat, milk and breeders to become more competitive in the market and industry. They also solve issues on production in community levels and are like extensions of PCC itself. Eastern Primary Multi-Purpose Cooperative is one of the firstly initiated cooperatives at the so called National Impact Zone (NIZ) in San Jose City.

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We were all tasked to come to Eastern PMPC for this last milk test day since there are a lot of samples that will be submitted and Mam Jilliene and Doc Chat need manpower.

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We arrived back at the Biosafety and Environment Laboratory at PCC just a few hours before lunch and we were tasked by sir Dadz to help him and the other docs to unbox materials, drugs and other equipments sent to them.

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In the afternoon, we were accompanied by mam Noemi in doing PCR for the detection or Trypanosoma spp.. This laboratory at PCC does molecular testing for detection of microbial pathogens such as PCR. Luckily for us, we have had a lot of experience in doing PCR since it is related to our theses.

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We left PCC headquarters at 5:30 am with Tata Naro and traveled for almost 3 hours to Gapan City, Nueva Ecija. We met one of the head officers of Parcutela  Cooperative, Mr. Godofredo, and he and his family let us into their home and prepared for us a delicious breakfast.

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Tata Naro then proceeded with his routine of diagnosing pregnancy for those served through AI, deworming and giving multivitamins to water buffaloes of all ages. He also taught us the proper way of administering these drugs orally and intramuscularly, respectively, but the most important thing we learned was the proper restraint of these large ruminants. Most of the farms we visited have chutes and holding pens but for those who did not have, restraint was achieved by tying the buffalo to a tree or post and having the farmer owner help in the restraint. Sometimes it needs more than two people to restrain these buffaloes. Tata Naro advised that for those aggressive buffaloes or those difficult to restraint, the farmers should just mix the dewormer with darak or rice bran and feed it to them.

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The vitamins we are essential nutrients or supplements for water buffaloes just like minerals are. They are important for metabolism and growth, reproduction and even health. In water buffaloes, the general concern of farmers are mostly on fat soluble vitamins particularly A, D and E.

Vitamin A plays an essential part for vision, immune system function and reproduction. Vitamin D, on the other hand is important for the metabolism of calcium and phosphorus and bone growth. Lastly, vitamin E is an antioxidant in cells and tissues and plays an important role for immune function as well. All these so called “fat-soluble” vitamins are stored in the liver and in adipose tissues so that daily supplementation is not needed. This is in contrast to water soluble vitamins which needs daily supplementation. In the case of water buffaloes, rumen bacteria supply the needs for vitamin K and B vitamins in most situations so they don’t need supplementation via injection with these vitamins. Tata Naro also carries in his car injectable B vitamins and liver extract which he sometimes administer to water buffaloes with non-specific anorexia , during convalescence from diseases and those with debility and showing general weakness.

Albendazole is the one used commonly by PCC technicians or veterinarians to deworm the water buffaloes with the exception of those pregnant, lactating and currently milked for production.It is a suspension and a broad spectrum endoparasiticide against a variety of nematodes, cestodes and even protozoa. It is labeled against Ostertagia ostertagi, Haemonchus spp., Trichostrongylus spp., Nematodius spp., Cooperia spp., Bunostomum phlebotomum, Oesphagostomum spp., Dictacaulus vivaparus (adult and 4th stagelarva), Fasciola hepatica (adults), and Moniezia spp.Unlike albendazole, ivermectin is also an ectoparasiticide. It is a prototype avermectin drug used in variety of species. At times, we subcutaneously inject ivermectin only in caracalves .It is approved for use in the control of gastrointestinalroundworms (adults and 4th stage larva), lungworms (adults and 4th stage larva), grubs (parasitic stages), sucking lice, and mites (scabies). It lacks data about publichealth safety and so is contraindicated in lactating animals.

Tata Naro also told us that it is not that rare to encounter trypanosomosis in farms in Nueva Ecija. He diagnoses them through pattern recognition and usually suspects it for those with clinical signs of emaciation, weight loss and abortion.And so, he carries a drug which is Isometamidium Hydrochloride (Trypamidium Samorin). It is a trypanocidal powder for injection against trypanosomiasis at 0.5mg/kg body weight in cattle for curative, but 1.0mg/kg body weight at three months interval for prophylaxis of animal trypanosomiasis. Tata Naro mixes it with sterile water for injection and also allows us to administer it intramuscularly.

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 Most of the time, Tata Naro let us to practice our skills and allowed us to administer all the drugs he would prescribe to the animals. He would just be the one to record them and do physical check-up and rectal palpation. His records were many which represent the many number of farms he would visit a day and log in his daily reports. For those caracalves without a registered number, Tata Naro would give them ear tags of which technique he demonstrated to us. Records and ear tags are important tools in keeping track of the production and health parameters even in smallhold farms and would help PCC monitor the animals in each area.

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In one of the farms we visited, we observed that the animals are alopecic most commonly in the head, neck and back area. We observed the animals closely when they were restrained and decided to take hair samples that we will bring back to the laboratory. When we looked under the microscope, we observed nits which are the empty egg cases attached to their hair that the buffalo  lice hatch from. Buffalo lice (Haematopinusaspp.)are considered the largest species of the genus occurring on ruminants but are not of great importance.

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For suspected cases of Surra which were diagnosed by Tata Naro through pattern recognition, we collected blood samples from the coccygeal and jugular vein, placed them in purple-top tubes filled with EDTA and transported them via ice box to the PCC headquarters to be processed in the laboratory.

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                Doc Chat gave us a laboratory request form to fill-up first before we can process the blood sample. This was a similar process that farmers or researchers undergo when they submit samples to the Biosafety and Environment Laboratory for processing. Currently, the laboratory performs fecalysis, blood parasite examination, serological testing (Brucella Test, ELISA for Neospora and CAE), molecular testing for detection of microbial pathogens (PCR, nPCR among others), and conventional microbiological testing (bacterial isolation and identification, antibiotic sensitivity testing, coliform count and total plate count).

            With the help of one of the assistants in the laboratory, Mam Noemi, we made blood smears of the sample and observed them under the microscope. We also centrifuged samples and observed them under the microscope but both methods showed negative results.

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As early as 5:30 in the morning, we are already on our way to start the first day of a series of Milk Test days. After almost two hours of traveling with Mam Jillian and Tata Naro, who were our companions from the Production Systems and Nutrition Section of PCC, we finally arrived at the very first cooperative we encountered for this externship which is Ayos Lomboy Dairy Producers Cooperative in Guimba, Nueva Ecija.

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Dairy farmers were already grouped there when we arrived and some were preparing milk samples for the CMT. Some milk samples were placed in gallons and tanks but the ones placed in four different tubes represent the four teats of the water buffalo and it provided more ease of testing. Mam Jillian made us demonstrate how to do the CMT  or “Cow-side Test” after introducing that it is a qualitative test  for detecting subclinical mastitis. We taught the farmers the CMT by the one Doc Chat discussed yesterday.

First, milk samples were drawn in each well of the CMT paddle corresponding to the quarter of the water buffalo. The paddle was tilted to ensure that all the milk was in equal amount. Equal amounts of the reagent solution were added and the paddle was rotated to ensure mixing. The paddle was then observed for a reaction which was graded as Trace, +1, +2, and +3 according to the viscosity. We reminded them to not mix the paddle for more than 10 seconds and to read the result quickly as it may dissolve after 20 seconds. For collection of samples in the future, we reiterated that they clean the udder and teats first, and do forestripping (voiding of the first milk acquired from each teat) at least three times.

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We remembered the one we read on the first day that this is a part of the services offered by PCC to help smallhold farmers. They offer these services as part of their extension support, education and information dissemination and training of technicians and farmers alike. The local farmers in Barangay Ayos Lomboy showed their gratefulness by preparing us a sumptuous breakfast.

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For the rest of the day, we asked permission from Sir Dadz to allow us to go with Tata Naro in his travels for the day. Tata Naro is a supervising livestock specialist of PCC. It is also Tata Naro’s job to extend the services of PCC to the local farmers even in far-flung areas. We traveled to Talugutug to Cabanatuan City to Talavera and then back to the Science City of Munoz so Tata can visit each and every carabao he intended to visit for the day. Most of the carabaos we visited were entrusted by PCC under the Dairy Buffalo Module. It is a service whereby PCC gives dairy buffaloes to qualified farmer-trustees, families or cooperative members who are capable of running a dairy farm business. It follows a Modified “Paiwi” scheme wherein the one entrusted by PCC will take care of the buffalo, maintain records and provide all necessary information required by the contract, and follow approved practices by PCC. The calves produced by the dairy buffalo will be co-owned by PCC on a 50:50 scheme. We observed this practice as Tata was tasked by PCC to receive half the payment from the trustee who sold one of their dairy buffaloes to the meat buyers. Farmers are allowed to buy PCC’s share in each calf or vice versa.

Artificial insemination (AI) is also one of PCC’s banner sevices which AI technicians render to farmers nationwide through their PCC centers which we also have in UPLB. It is a great reproductive tool ued to produce buffaloes with better milk productivity without the hardship of transporting a bull. As part of technical assistance and extension which is free of charge provided by PCC, Tata Naro also checks if AI was successful and the caracows became pregnant by doing rectal palpation. He also allowed us to administer multivitamins (Vitamins A, D, E and K) and a dewormer (Albendazole) to a caracalf. We visited many farms and Tata Naro provided the farmers with information on animal reproduction, nutrition and health and other areas he was knowledgeable of. We learned that he does regular on-site assessment and field visits to respond to the farmers’ needs and they call him Doc Naro. Our duration on each area varied depending on the situations on that area and also on our merienda time. Tata Naro again contacted Sir Dadz and scheduled a trip for the next day for us and sir Dadz allowed us to go again with him. We arrived at the PCC headquarters in the afternoon and sir Dadz called us back up to the Biosafety and Environment Laboratory.

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The laboratory is where farmers and other interested parties submit samples from farms for analysis and it is one of the services of PCC. They help diagnose diseases whenever they can. And so, whenever we were at the laboratory, there are lists of buffaloes or farms where samples are collected. This time, we did the Flotation technique for fecal analysis. We added water to tubes filled with fecal samples which were previously prepared by sir Dadz, and placed coverslips at the top. We waited for about 20-30 minutes to allow ova to float such that they will adhere to the coverslip when we remove them. In contrast to the Sedimentation technique, no staining of methylene blue was done. This technique is useful for observing for the presence of strongyles, Strongyloides spp. and also coccidia. The only ova we detected was that of Strongyloides spp.

We had been preparing for the start of this 5-week externship since last year. We had breakfast in our apartment and we’re ready to go. We started walking from Roseville Subdivision near the Welcome arch to San Jose City to the Philippine Carabao Center National Headquarters and Genepool at around 7:30 in the morning. It was bright and we were very excited about the things we are about to learn.

We were instructed by our contact, who we knew by the name of Sir Dadz as introduced virtually by our former senior clinicians, to log in at the gate and head to the Human Resource Department (HRD).

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As we entered the building where the HRD is located, we were again asked to log in the visitor’s logbook and were briefly confronted with a short introduction to the Philippine Carabao Center.

One of the texts read:

 “The Philippine Carabao Center, operating as an attached agency of the Department of Agriculture, is mandated to conserve, propagate, and promote the Carabao as a source of milk, meat, draft power and hide to benefit the rural farmers.”

With its vision:

“ A premiere research and development institution propelling sustainable growth of the livestock industry.”

And mission:

“Improve the general well-being and competetiveness of the livestock industry stakeholders through animal biotechnology and technology development, technology dissemination and knowledge resource management, active private sector participation, livestock-based enterprises, and policy reforms to sustain development of livestock enterprises, thus ensuring socio-economic empowerment for nation building.”

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We then headed to the third floor where the HRD is located. We saw Mam Helen, our contact from the HR department, going down the stairs and hurrying to attend the Monday ceremony and meeting and instructed us to re-meet at around 9 am. We toured ourselves around the building instead. We were in awe with the huge facilities and beautiful rooms of PCC.

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We passed time at a sofa and table waiting area just outside the HRD and Executive Director’s Office. We read magazines and official newsletters of the PCC and told stories of each other’s vacation. The magazines were all filled with new trends in the agriculture industry and happy farmers whose lives were improved. They were pretty inspiring.

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Mam Helen arrived at 10 am and spared just a short time since their meeting was not yet adjourned. She asked for our requirements (Acceptance Letter, Curriculum Vitae, and Parent’s Letter of Consent) and gave our respective On the Job Trainee IDs. She then passed us under the supervision of sir Dadz and said to meet him at the lobby downstairs. Mam Helen wished us luck and more learnings and we headed downstairs. We just can’t wait to start our duty at PCC. The people seemed amazing.

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Downstairs was a whole lot more magazines and books to read . PCC was not only good with helping people, they help create more amazing stories too.

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Sir Dadz arrived at the lobby at 11 and apologized since he also still have important matters to attend to at their meeting and said to meet us later again after lunch at around 1. But, before we went to grab lunch at the nearby carinderias, we rode a jeepney to Barangay Bagong Sikat to find the Bicycle Rentals since it is more economical and ecological as well to use these bicycles in going to PCC headquarters and especially Gene Pool from our apartment. The Bicycle Rentals was almost closed since bicycles were banned at CLSU campus and unfortunately, we cannot use them to take a leisure trip there. Thankfully, the owner of the Bicycle Rentals was a veterinarian and opened his bicycle renting services to us. We each got a bicycle and drove again back near PCC to have our lunch. We were very much in love with our bicycles.

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We arrived at the laboratory and sir Dadz gave us a short tour around the PCC Biosafety and Environment laboratory, orienting us the proper working areas, machine uses and disposal bins. Every thing inside this laboratory is expensive but is of great help to the many researches conducted here. I remembered one of the posts we read earlier before going up the stairs which read:

“The Philippine Carabao Center aims to be a premier research and development institution for the sustainable growth of the livestock industry”

We were overwhelmed with such beautiful and expensive equipment but at the same time are hyped to use them.

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Sir Dadz also oriented us the proper attire at the laboratory. He then tasked us to centrifuge the serum samples at 1400rpm for 2 minutes to thaw them in preparation for the Rose Bengal Plate Test (RBPT) or Rapid Plate Test (RPT). We have only encountered this test in textbooks before and now we are doing them.

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The RBPT is a simple, rapid slide-type agglutination assay which is aimed at detecting antibodies of Brucella species (more commonly for the detection of B. abortus, B. melitensis and B. suis) in animal sera such as ruminants, equidae, suidae, camelidae and carnivores, both wild and domestic. For this day, we tested sera from water buffaloes.

The reagent is a pink suspension consisting of a Suspension of Brucella abortus biovar 1 Weybridge strain No 99, inactivated by heat and phenol and coloured with rose bengal stain in an acidified buffer.This antigen is available commercially, for veterinary use, and stored according to the supplier’s instructions. Before starting with the technique, Sir Dadz made use of a weak positive serum as control to show as what a positive reaction would look like.

We then proceeded with the test by adding a serum sample (0.03 ml) and mixing it with an equal volume of antigen on a white tile to produce a spherical area approximately 1-2 cm in diameter. We gently agitated the mixture for four minutes at room temperature, and then observed for agglutination with good lightning. We will count any visible agglutination as positive but agglutinates revealed after 4 minutes ± 10 % will not be taken into consideration. Fortunately, none of the sera tested positive.

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After that, we were introduced to another routine test to check for Gastro-intestinal parasites which is very familiar to us, Fecalysis. We made slide preparations of the fecal samples after sedimentation technique. This technique is a qualitative method as oriented by Sir Dadz earlier and he also said that the eggs we should expect to observe under the microscope are the “heavy” eggs which will sink to the bottom after sedimentation. These are the ova of amphistomes, paramphistomes and Fasciola spp. He also warned us of the artifacts and pseudo-parasitic particles such as pollen grains which we may mistake as parasite eggs.

We obtained approximately 2-3 grams of feces and placed it into a container. We mixed it with 50 ml of tap water, mixed it thoroughly with a spoon and passed it through a strainer and into another container to remove grass and other large debris. The fecal suspension was then transferred on a 50-ml conical-bottom disposable plastic tube. The sediments were allowed to settle and then approximately 40 ml of supernatant were discarded. The sediments were resuspended with new tap water and were allowed to sink for approximately 20 minutes. The supernatant was again discarded very carefully and the sediments were acquired using a glass pipette and transferred on a microscope slide. The sediments were stained by adding a small drop of methylene blue. The slide preparations were covered with coverslips and we checked them under the low power objective of the microscope; the only egg we observed was that of the liver fluke, Fasciola spp.

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We prepared California Mastitis Test (CMT) reagents after performing the two laboratory tests in preparation of the 3 consecutive milk test days starting Tuesday around the different barangays and towns of Nueva Ecija. To instruct and refresh our memories about CMT, we headed to Doc Chat’s room so she can discuss it to us.

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We later on learned from Doc Chat that the reagent we were diluting constituted a anionic surface -active surfactant and bromothymol blue as the indicator dye. We prepared it by adding 1 pint of the reagent to 3.78 liters of distilled water to dilute it. She also discussed how the reagent works. It makes the milk viscous after mixing (1:1) by lysing the somatic cells, if present, and denatures any DNA. The higher the somatic cell (WBCs) count, the higher is the viscosity of the mixture. The number of somatic cells in milk increases when the number of pathogenic bacteria such as Staphylococcus spp. and Streptococcus spp. are present causing mastitis.