Day 20: Rumineyshennation

Today, we attended a proposal of a product by Mr. Aner Yacobi entitled SCR Fertility and Cow Welfare. He proposes to PCC staff, veterinarians and heads of offices an electronic ear tag, yet to be released in the country, which measures the activity of a cow to detect estrus as well as to measure the rumination time of a cow which correlates to its health and nutrition. Mr. Yacobi said that cud chewing is a factor where nutritionists in their country heavily rely on to assess the nutrition of the animal. One of their products already released is a collar type which measures movement of the neck. However, the only problem they encounter with this design is strangulation of the animal.

Mr. Yacobi started by saying that “If you can’t measure it, you can’t manage it.”. He is referring to the measurement and detection of estrus to allow rebreeding and make an efficient breeding program. He says that the product helps optimize herd health, fertility and nutrition which are keys for profitability. The product sends data to a modem and then sends the data to a computer which can be placed in an office. Data can easily be managed and read since they are immediately presented in tables and graphs. However, training is required before interpretations can be done. The product has never been tested to buffaloes yet. PCC is willing to conduct trials to detect the accuracy of such products to their own herd.

Just before lunch, we taught Novy how to extract DNA from some blood samples. After lunch, we went to observe Mam Noemi who was examining three disposable plastic dishes containing a fungal culture she made from paddy straws. She aims to isolate the causative agent for Degnala disease which is Fusarium spp.. She also made an enriched culture. It was quite difficult to acquire fungi from the culture and place them on glass slides for observation under the microscope unlike for bacteria. We then decided to use Scotch tape test to acquire some fungi from the plates. We stuck to tapes to the glass slides and slightly stained them with methylene blue. Based on our examinations, the cultures contain not Fusarium spp. but Aspergillus spp. based on morphology. Aspergillus spp. is a ubiquitious fungi and is an opportunistic pathogen. We disinfected immediately the slides and materials we used and then went on to process the purulent fluid we collected yesterday.

With the help of Mam Noemi, we inoculated some of the purulent fluid into three plates containing nutrient agar (NA). NA is a general purpose medium which supports the growth of a wide range of non-fastidious organisms. We then stained some of the purulent fluid on a glass slide using Gram stain (1 minute crystal violet, 1 minute Lugol’s iodine, 10 seconds alcohol and 30 seconds safranin). We observed some gram positive cocci in few numbers and suspected that they were just contaminants. We will wait for the culture results and then stain again later to observe for the bacteria which will grow.

Day 19: Milktasteagen

Today was another Milk Test Day and it was Novy’s first time to attend the activity so we guided her aside from Doc Chat’s tutorial. We left the headquarters at 6 in the morning together with Doc Bjet and Doc Jonathan. We arrived then in one of the cooperatives supported by PCC, Pulong Buli Primary Multi-Purpose Coperative in Sto. Domingo, Nueva Ecija. We immediately started with the milk testing using the California Mastitis Test. All the negative samples for CMT were placed in milk cans to be pasteurized and sold as dairy products. Some amount of the negative samples was placed in 50 ml tubes to be submitted at the Meat and Milk laboratory to be analyzed for contents. Mam Paulen said last week that those cows yielding the best quality milk based on the milk analyzer will be used for breeding to improve the production of each dairy farm and in other words, the cooperative itself.

There was a meeting after the milk testing and Doc Bjet asked who were interested to avail carabull or caracow loans from PCC. After the meeting, we accompanied Doc Bjet in going to Saranay or LBRAF where the buffaloes for loan are being housed. Doc Bjet who was a roaming veterinarian in PCC was also responsible for checking some of the buffaloes for loan and transacting them to those interested to avail.

In the afternoon, we headed back to Sto. Domingo because a farmer called Doc Bjet for a case in his farm. According to the farmer, his caracalf has a swelling in the abdomen and Doc Bjet suspected it was umbilical hernia. He instructed us to prepare for a surgery and so we wore our boots and gloves, carried the surgical instruments, drapes, drugs and other paraphernalia to the pen after it was cleaned by the farmer.

Doc Bjet immediately saw the swelling since it was already big, the size of a medium melon. Doc Bjet palpated the swelling and it was soft and feels like a fluid inside is present. Doc Bjet suspected it was not umbilical hernia but navel ill or omphalitis. It is a condition wherein the navel or umbilicus of a caracalf is infected by contamination of bacteria which is usually found in the environment. This condition is common usually soon after birth and is localized. However, Doc Bjet said that such cases should be attended immediately since the swelling may serve as a source of infection leading to septicemia. The caracalf was 1 month old and is still active and not feverish which indicates a good prognosis.

 With the help of the farm owners, the calf was restrained by tying its neck to a fence and tying both the front legs together as well as the hind legs. When the caracalf was already in lateral recumbency, one of the owners was holding it down at the head region while some of us were restraining it near the tail region. The umbilicus can better be seen and it was enlarged, somewhat painful upon palpation but is not draining any purulent material through a fistula. Doc Bjet inserted an 18-gauge needle connected to a 50 ml syringe through the swelling and began aspirating. Doc Bjet aspirated milky white, purulent fluid from the swelling and confirmed that it is omphalitis based on his experience in the field.

                Doc Bjet decided to open the swelling to drain the pus faster and to clean the inside of the swelling. Previously prepared povidone iodine solution was sprayed onto the surface of the swelling and the swelling was shaved with a blade to remove the hairs. Using a different blade, a #20 scalpel blade, a small incision was made on the swelling and pus was drained to a steel basin. Approximately 600 ml of pus was drained by continuously massaging the swelling. When a maximum amount of pus was already drained through massage, the inside of the swelling was flushed with povidone iodine solution using a syringe with an attached sterile cut AI catheter to reach further inside. This technique is common and we’ve learned this during our small ruminant duty where povidone iodine can be used for uterine lavage to clean the uterus of bacteria. Povidone iodine is an effective antiseptic and is effective against a wide range of bacteria. Therapeutic irrigation or lavage was done several times until no pus can be observed going out with the solution.

The inside was then sprayed with chlortetracycline hydrochloride, a tetracycline antibacterial effective against many opportunistic bacteria both gram-positive (Staphylococcus spp. and Streptococcus spp.) and gram-negative (Bacillus spp., Clostridium spp., etc.). Next, a screw worm aerosol was used aas a wound spray to prevent myiasis or the infestation of the wound opening with fly larvae. Its active ingredient is permethrin which is an insecticide that kills flies and maggots that come in contact with it.  Intramuscular injection with an antibiotic, amoxicillin + gentamicin, was also done since topical antibiotics are not that effective for treatment alone and in this case was only a supplement to intravenous antibiotics. Tolfenamic acid was also administered intramuscularly in a site different from the first intramuscular injection as an anti-inflammatory drug and analgesic. Lastly, tetanus toxoid or tetanus vaccine, an inactive vaccine used to prevent tetanus, was injected at the gluteal muscles. Doc Bjet said that tetanus toxoid must always be given when doing surgery especially when wounds are being created because tetanus (caused by the bacteria Clostridium tetani) is common and can be deadly. Doc Bjet said that he will go back to the farm to administer antibiotics for the following days and to check the prognosis of the condition. We brought some of the purulent fluid back to the laboratory and stored it for culturing and morphological staining.

Day 18: HayleyBaley

For today, we were assigned to Tata Mar together with Novy, a new veterinary student intern from NVSU. We accompanied Tata Mar in going to dairy farms and extending the services of PCC such as deworming, vitamin administration and pregnancy diagnosis. We guided Novy in our usual routine especially in drug administration as well as in record keeping. Tata Mar taught her rectal palpation.

We also dewormed all the caracalves in one dairy farm with a new drug we encountered in the field which is doramectin. Just like ivermectin which we were accustomed to using, this drug is also an avermectin antiparasitic agent. Based on its label, the drug is usually indicated for use as an injectable in cattle and swine and as a topical in cattle. It is indicated for the treatment and contol of the following endo- and ectoparasites in cattle: roundworms (adults and some fourth stage larvae)—Ostertagia ostertagi (including inhibited larvae), O. lyrata, Haemonchus placei, Trichostrongylus axei, T. colubriformis, T. longispicularis, Cooperia oncophora, C. pectinata, C. punctata, C. surnabada (syn. mcmasteri), Bunostomum phlebotomum, Strongyloides papillosus, Oesophagostomum radiatum, Trichuris spp.; lungworms (adults and fourth stage larvae)—Dictyocaulus viviparus; eyeworms (adults)—Thelazia spp.; grubs (parasitic stages)—Hypoderma bovis, H. lineatum; lice—Haematopinus eurysternus, Linognathus vituli, Solenopotes capillatus; and mange mites—Psoroptes bovis, Sarcoptes scabiei. The manufacturer also states that doramectin protects cattle against infection or reinfection with Ostertagia ostertagi for up to 21 days.

While we were about to leave and go to another farm, we noticed a tricyle driver delivering some grasses to feed to the buffaloes. The grass was not the usual Napier grass we see that were being fed to buffaloes. The grass was barnyard or cockspur grass (Echinochloa crus-galli), a type of wild grass in the country which was previously classified as a type of panicum grass. It was one of the worst weeds and can be found along roadsides, ditches, along railway lines, in disturbed areas such as gravel pits and dumps, riverbanks and the shores of lakes and ponds. It was removed by the farm boys who delivered the grasses since it was a nuisance and for buffalo feed as well. These  warm-seaspm grasses were documented to have been used in some countries as cattle fodder throughout the year and is sometimes cultivated for that purpose. Some authors said it was suited for silage but not for hay. Tata Mar said that while it can be fed, it provides lower quality of fiber and nutrients as compared to Napier.

The next farm we went to was at the top of a hill overlooking onion farmlands and it was very windy and has a breathtaking view. This farm is also an integrated farm with swine pens and free range poultry. The chickens were also beneficiaries of the leftover feeds from carabao feed according to the farm manager which is an advantage since nothing or minimal feed is wasted. We also noticed hay which were compacted, bundled or baled. They used a large round hay baler, a farm machinery used to compress and roll cut and raked hay. This method was done for the ease of handling, transport and storage of hay. The cylindrical hay was also bound by a wire which Tata Mar said to dispose of properly since the buffaloes may accidentally ingest them.

After the caracows underwent pregnancy diagnoses through rectal palpation, we noticed one caracow which has horns curling back to its head and nearly puncturing it. Tata Mar then asked for a steel saw and began to saw the distal 1/5 of the horn. After sawing the tips of the horns, Tata Mar also asked for a whetstone to remove the sharp edges of the horn and prevent future damages. The procedure was quite fast and easy and did not require prior anesthetic administration and post-operative wound management since there was no bleeding.

Day 16: Ubayxtrakshen

Sir Dadz told us that we still don’t have a scheduled assignment for today since most of the samples have already been tested (i.e. fecalysis and RBPT) and processed (i.e. DNA extraction) last week. We decided to just review some of our old notes for the mean time. Mam Jona from the Meat and Milk laboratory of PCC texted Sir Dadz and telling him to get some blood samples from their laboratory. James and I went to the laboratory and she handed us an ice box containing some blood samples in plain vials. Based on the labels, these were from the Ubay Stock Farm of PCC in Bohol.

Also called Ubay Agri-Park, is one of the oldest and also the largest government livestock facility in the country. Based on the stories of some of the PCC staff, the facility also houses different farm animals aside from the carabao which are cows, horses, goats and poultry such as chickens, turkeys, and ducks. Following one of the core principles of PCC, the facility also helps in developing the livelihood of farmers while also catering to tourists who would like to have some fresh milk and other dairy products.

            The facility has sent some samples for screening of some diseases. So for the whole day, all of us extracted the DNA from all 105 samples. We followed our usual protocol for DNA extraction:

  1. In a 1.5 ml microcentrifuge tube (MCT), 1000ul of NH4Cl were added to 500ul of buffy coat/whole blood sample.
  2. The mixture was mixed by pipetting/vortex mixer, and then centrifuged at 14000 rpm for 1 minute.
  3. The supernatant was discarded.
  4. White pellets after centrifugation were observed. Then, steps 1-3 were repeated.
  5. 1000 ul of Cell lysis was then added, the mixture was vortexed to dislodge the pellet, and then it was centrifuged at 14000 rpm for 1 minute.
  6. The supernatant was then discarded by pipetting.
  7. 300 ul of Nuclei lysis solution was added.
  8. The mixture was again vortexed for at least 30-40 seconds.
  9. 100 ul of Protein precipitate was added, and then the mixture was vortexed until it was homogenized.
  10. Centrifugation was also done at 14000 rpm for 10 minutes.
  11.  While waiting, new 1.5 MCT were prepared with each containing 500 ul of isopropanol/propanol.
  12. The clear liquid after centrifugation of samples was transferred to the new 1.5 MCT and then mixed with inversion.
  13. The mixture was then centrifuged at 14000 rpm for 1 minute.
  14. The supernatant was discarded.
  15. 500 ul  of 70 % Ethyl alcohol was added.
  16. The mixture was then centrifuged at 14 000 rpm for 1 minute.
  17. The supernatant was then discarded.
  18. The tubes were air dried for at least 30-40 minutes without overdrying the DNA.
  19. Rehydration was then done by adding 30 ul of DNA rehydration solution.
  20.  The DNA samples were incubated at 65 degrees Celsius for 10 minutes and then stored in 4 degrees Celsius.