Hannah Asquith – 2023 IWU Freeman Asia

Lab work for the Week

Monday was slow I headed into lab bright and early in the morning to find that my advisor was working from home and that my fellow intern also would not be in lab. So I decided to pour some plates from the previous week and take photos of out antagonistic plates to right down some observations in my notebook. after ending early on Monday, Tuesday finally had some work for us to get done. This included going out into the filed and collecting some root and leaf samples. We collected from rice patties with a nitrogen value of 0 as well as a high nitrogen value patch. We collected 20 leaf samples and four root samples which we would process the next day. We would pull one plant and cut 5 leaves off of the plant and place them into a sterilized tube. And for the roots once the plant was one of the ground we had to shake/pull the mud off of the roots and place them into a 100 ml conical tube. When collecting the roots the area in which we will be isolating from is the rhizosphere, this is the area right outside of the roots (shown below).

This is where we are most likely to find endophytes that grow in the root systems. The next day on Wednesday we had the task to disinfect and plate the leaf samples. The procedure I found during my research to disinfect the leaves and roots was as the following:

1. Rinse with sterile dH2O
2. Dip in 7% ethanol for 1 minute
3. Dip in 2.5% Sodium hypochlorite (Chlorox) for 4 minutes
4. Dip in 7% ehtanol for 30 seconds
5. Rinse three times in sterile dH2O

The set up or the disinfecting and plating process looked is shown below.

After the disinfecting process we had to cute our 4 leaf pieces into 2 and place them on two PDA plates and 2 NA plates. During plating we also had to injure the leaves by making small incisions on the sides to induce infection! Once that was completed which took the entire day by making 40 plates each, we placed the NA in the 37 degrees C incubator and the PDA plates in the 28 degrees C incubator. Thursday was yet another busy day filled with mainly making plates for the next day of experiments. We made 40 plates of both NA and PDA as well as about 40 tubes of PDA slants and NA Slants. Friday was a bit more exciting with Vincent returning and us being able to see some of the cable bacteria that grew in the slides for a week!

The cable bacteria quite literally look like cables or to me they look like hair under the microscope! These do help with reducing methane emission in rice soils however, they do make the soil quite acidic with the amount hydrogen they produce.

next we had to analyze our results from our leaf plates! Most of them seemed to grow either a fungus (on the PDA) or a bacteria (on the NA) below are the results!

We did not have enough time in lab this week to streak for isolation but we will complete that task next week.

The last activities we did on Friday is go out into the field again with Vincent to collect sample from the high nitrogen levels patch. The images of the cable bacteria came from a soil sample of high nitrogen. The samples from low nitrogen had no cable bacteria. So we scooped a huge bowl of mud and sifted the mud using a strainer and then put it in a conical tube! It was very hot that day and we were all sweating quite a lot, so after sampling we all went for an iced coffee to end our work week!

Weekend Activities!

This was another slow weekend. We started it on Friday night where we went to a Karaoke place called LBeats. We sang so many songs, its was fun! Then on Saturday the group split up with me and Julia hanging around Los Banos due to an errand I had to run. The idea was that we needed to find Bayluscide which is an apple snail control that is only sold here, and my dad wants some for our taro patches back home. When I tell you it was journey getting there, it was. We started at a café to get some food into our system. I had some waffles and a brown sugar latte, Julia got a strawberry latte and we both thought it was quite interesting. Next we went to find a charger because Julia’s phone was about to die and she had the google maps on it. After that we headed into a part of town that we both haven’t walked in. The streets were busy and after another 20 minutes of walking we stumbled into the small farming supplies store. The store was so small it seemed as though it could only fit three people. I was lucky enough to get the last box of 10! Next we decided to hang at one of the smaller malls where Julia and I sat at a coffee shop reading our palms for like 2 hours. Then we decided to get some messages in the message chairs here.

Once Julia and I finished our we decided to start the trek back. After walking for a bit we were hungry so we stopped at a somewhat fancy restaurant called Bonitos. It felt so nice to sit down in an AC room for a bit, plus we were very hungry. We got our food and enjoyed our time. When we were about to leave there was a huge crack of thunder followed lighting and some heavy rain. We were stuck in the restaurant for a bit and then when the rain lightened a bit we made the decision to head out. It was drizzling a bit but the thunder and lighting were heavy. It was quite beautiful though because we were walking during sunset so when the lighting flashed the whole sky turned purple. Sunday was yet another lazy day where Jessica N. Julia and I went to a café to do some work and had some yummy treats. Till next time folks! 🙂

Lab Work/ Week Work

Due to my four day weekend I didn’t start my week until Tuesday which surprisingly wasn’t a lab day! Instead we were able to go out into the rice fields to get a taste of how the farmers work so hard everyday. Our day started at 6:30 in the morning with a boot fitting! Then we headed out to the patches for our day of work. We were put into assigned groups and I was partnered with a very kind older woman named Ramil. She told me that she was here for the day just for fun and that she isn’t an intern. Ramil has 7 hectares of rice farm land which she manages, and she used to work at IRRI in the social sciences department. She also worked at UP LB as a professor for the social sciences. Our group started with preparing grids for manual rice planting. I had fun with this task because all we had to do was drag a wooden fence looking at things across the mud. The rice patches are very similar to kalo patches, I didn’t feel any difference while working in them, so I had a very easy time moving through the thick mud! Next I rode on the planter machine which was a small tractor with floaters and a planter on its rear rather than a tiller. It was smooth and easy. I can see why they use it so much. We were then instructed to get into the same patch that we made our grids in to start manual planting. Just like kalo all that you do is plant the plant in the intersection of the lines. Working on the farm reminded me of home and when I go out to the lo’i with my dad. Next was a hydro-tiller which was like a push lawn mower but in the water tilling the mud. Then we got to use a carabao (swamp water buffalo) to till a more muddy patch! She was very sweet but walked very fast (the cow). Next was another tiller, not too sure what that one did exactly but they ran super fast and I feel like I am jogging in the mud with this machine. The last machine was a hand held planter which is shown below. This one wasn’t too fun, it was heavy and when I got it I got stuck for a second and the helper had to run over to get me unstuck (how embarrassing).

Then the second to the last station was built like a pa (or banks) for the rice to be planted on. This technique is for the experiments done on rice, they have these banks that run across the entire width of the patch and those lines are where you can plant rice. This helped because you can have more than one variety of rice or even stages of rice right next to each other. One of the instructors also told us about the golden apple snail! He said the late president’s wife visited a farming region in Indonesia and saw that they had these snails being cultivated and eating on a daily basis. The snails were grown in wet environments similar to rice patties and they are a great source of protein. So she decided to introduce them to the Philippines so that when farmers are out all day they can have a snack on the farm! So many apple snails are now found on rice patties without needing to be introduced. (In Hawaii they are quite invasive, and I forgot to ask the farmers here if they are also annoying and invasive). Lastly we got to watch some drones fly super high in the air which help farmers scatter fertilizer as well as spray pesticides. Once the day was over I was tired and ready for a nice shower and my bed. The next day I was back in the lab after my long weekend and the start was very slow as we weren’t doing much this day. On Wednesday the only activity in the lab was to prepare freezer stock for the endophyte starins I had already revived. Instead of storing them in skim milk we stored them in NB and Glucose this time. The procedure was simple, all I had to do was pipette some of the culture broth I made last week into some of the freezer tubes along with some 50% glucose I also prepared a month ago. My advisor had to finish his abstract for this project by this weekend so he was a bit busy. But the next day a new group of interns came to the lab. To our microbiome team we got a new team member whose name is Dane. She is a botany student from UPLB college here at IRRI for a one month internship. We made pleasantries and got right to work with some antagonistic testing. This is where we split the plate in two and spread our endophyte culture from last week on one half of the plate (using a glass spreader) and on the other side of the plate we place a spore of the fungus. Then we wait overnight to see if the fungus has either taken over the area of the endophyte or if the endophyte strain shows a resistance or an antagonistic characteristic to the fungus. Next I had to attend our lab meeting where the new master student presented her thesis. The day ended on a slow note. Finally Friday hit and I was surprised with the announcement that we will be going out with another lab to collect some soil samples for their project. That is where we met Vincent. He is a PhD graduate conducting his post research in cable bacteria. Cable bacteria are able to conduct electricity through oxidation in the soil of rice patties. They decided to collect samples from high nitrogen patches and patches with no nitrogen to see what the cable bacteria prefer. We also collected samples from areas with more clay and sand in the soil to see if that also makes a difference. The collection of the soil is done by pushing a glass with both ends open into the soil then placing a cap at the top to suck the soil stuck in the glass out of the patch. Once out of the patch you must place a cap at the bottom of the jar to inhibit soil from escaping. Once we have the sample we head back to the truck and collect two different samples from that soil tube. One is from the top where there is oxidized soil and the other is from the bottom of the tube where there is no oxidized soil. When looking for cable bacteria in the unoxidized soil we wanted to collect roots that have an orange color to them, indicating iron in the soil and some sort of oxidation occurring. We were collecting samples the entire morning so after lunch it was back into the lab for us! Dane and I had to make more plates for the week of endophyte isolation to come. While that was in the autoclave we made sure to check on our antagonistic plates from the other day to see what the results were. We were checking to see if any of the strains showed resistance to the fungus. We did notice that only two out of the five strains showed a clearing from the fungus which meant that they are antagonistic. These two strains included In-b-590 and In-b-714. Next Vincent returned to our lab with his soil samples, he offered us a chance to learn how to prepare slides for bacteria. The procedure was simple, we had a slide with a square cut out from the middle which is where we placed our mud samples. You pack in there pretty well using a spatula, and wipe the edges using some tissue. Next you place some water on the slide over the dirt using a syringe and then put a slide cover on top of the dirt slide to keep air out and allow the bacteria to move around and live. Next we placed them in some large conical tubes and put them in a box on the counter till the next week. Vincent brought in his own microscope to show us what one of the rice roots looks like. The root looked very yellow under the microscope and had many blemishes on it. I could not see any bacteria in the rhizosphere which is what we were supposed to be looking for. And that concluded our week in the lab!

Weekend Activities!

This Saturday we took things a bit slow. The girls and I woke up somewhat early to head over to the market where vendors are selling food as well as some other goodies! Julia and I were stopped by a sampler who gave us some pesto pili nut spread to try, we did end up buying a jar each of that spread to share with our families when we return. Next we looked for some food which was not a difficult task because there were just so many vendors selling food, we decided to try some bread from a nice lady. Then one of Jessica’s Ate’s from the lab came and brought us to her university’s cultural food event. There was nothing vegetarian but the others sure did enjoy the food! (plus it smelled amazing). Next Julia and I got some coffee from one of the market vendors and sat under a tree and just talked for a while. After finishing our coffees the others met up with us and we ventured into town to find a store to buy some groceries from. We made a detour for a craft store where I bought a cute little glass jar to store my shells in. Once we finished our shopping, we all decided to head back to IRRI to cool down. Later in the day around 3 pm the Jessica’s came knocking on my door begging to come and play volleyball with them, so of course I dragged myself out of bed and went down to the field to be active. Volleyball was fun, the field (as like the rest of IRRI) has a beautiful view of the mountains and being outside is always nice. The grass was a little wet, so my feet got a bit dirty during playing. There were also some puff mushrooms peeking through the grass! Later that night we decided to go to a Korean BBQ but I was quite tired so I just stayed back for an early night’s rest. However, I did meet up with Julia after she came back for a movie night and to make out little glass jars. We both bought those little glass jars, and we both had some shells, so for a bit we sat and filled our jars with the shells we collected from El nido.

Sunday wasn’t that much different, one of our slow weekends I guess. But we did decide to head over to Calamba which is the next town over. There is a mall and movie theater there where we got to be in the AC for the rest of the day. We looked around at some of the different shops but ended up watching Mission Impossible. Nothing too exciting but we have had many busy weekends so it felt nice to just cruze a bit this weekend.

Anyways, till next time!

Below are the Antagonistic tests of the five endophyte strains against a fungus. If a clearing appears that means that the endophytes is resistant to the fungus and is indeed antagonistic.

Sorry for the late post,

In the lab…

This week started off to a bit of a slow start with presentations happening on Monday and Tuesday. Monday starting around 9:45 am, some speakers came to talk about some of the projects being done at IRRI. To start there was a talk about IRRI’s mitigation and some of the developments as well happening correctly. Then my on site advisor Dr. Van talked about the project I am working on in the lab during my stay here. She talked about plant microbiomes and plant rates that correlate with low methane emissions. After a few more talks there was a break in which I ran up to our lab to put away the NA plates I made last week and to take pictures of the results from the salt resistance test, the nitrogen detection, and phosphorus detection. On the Pikovkaya’s plates there seemed to be no clearing which would make sense since we were missing the ingredient for the detection of phosphorus. For the Jensen’s Medium there were also no clearings which meant there is no nitrogen fixing bacteria in these endophytes. The salt resistance test was very interesting though. Most of the endophyte strains selected were resistant to salt and grew nicely on the plate. The two strains who were not resistant were In-b-714, and In-b-24. The data for In-b-24 is inconclusive due to the fact that we are unable to properly grow colonies in the tubes. When looking at the salt resistance plates I am looking to see if there is growth in the spots that I directly pipetted my colonies and NaCl solution on. For In-b-714 there was no endophyte growth, only the formation of some salt crystals on the plate. For the rest of the day on Monday I attended more of the talks that were held for science day. On Tuesday my supervisor returned to the lab so we were able to get a bit more done. He looked at the plates I had made a week ago and agreed with my conclusions. Later that morning we headed out to the lower land field to collect some leaf and root samples for the isolation of ‘wild’ endophytes. It was hot and humid and the smell of unoxidized dirt reminded me of the days I spent with my father in the taro patches at Kealia. It was nice to have that feeling again no matter how sweaty I was. From each plant we picked five leaves, and placed an entire root system into some conical tubes and let them rest there until we are ready for the disinfecting and isolation process. On Wednesday I was tasked with preparing a lab progress presentation for our lab meeting the next day. I included all the results from the tests that I have already completed as well as the plans of my next steps. During these presentations in the lab meetings the other researchers are welcomed to stop your presentation and make comments on your work, and sure enough that did indeed occur. They mentioned that I should be more specific on how exactly I will be plating the colonies and they decided we should try to acquire and characterize as many endophytes as we can during my time here. Dr. van also mentioned that we should test the other frozen endophyte strains that haven’t been stored in skim milk, as well as test the new endophytes in skim milk. Once we were finished with our meeting it was the end of the day and we were free to go home. I took my four day weekend this week so I missed the lab on Friday. More to come about the lab in week 6!

THE WEEKEND!!!!!!!

So for this weekend all five of us as well as Chrishana and Semaj, took a four day weekend in order to visit the island of Palawan and stay in El nido. The Los Banos group left our dorm on Thursday at 11:45 pm to reach the airport which is two hours away for a flight at 4 am. Once on the plane, the ride was a little less than an hour and we landed in Puerto Princesa where we then drove for almost 5 hours to El nido. Once reaching El nido at around 10 in the morning we checked into our hostel called the outpost and hung around talking to each other and enjoying the scenery. Later that day me and Julia tried to swim out to where Sam and some of the others from FNRI went but it was low tide and there were weird things in the water that we don’t have in Hawaii. When I was walking there were some crunchy things in the sand, there were also holes which could have been made by some sort of worm, and there was seagrass. The sea grass was gross, you could be swimming and then all of a sudden get touched and of course I freaked out. After our attempt at swimming we sat at the hotel’s bar and restaurant for some dinner and later headed to bed to prepare for our big day! On Saturday as a huge group we made sure to book for an island hopping tour which started at 9 in the morning. I made sure to get up early and have some avocado toast in order to prepare for the activities of the day. At around 8:55 am the tour guide started to yell around the beach for all of us to meet there so we could head over to the boat. Early in the morning the tide was low but steadily coming in. The boat was anchored not too far from shore but about a 5 minute trend through the warm ocean water. Once on the boat we had to put on our life jackets to make sure none of us fell in the water. The only size the crew had were Large’s so as I was sitting in the boat I was being choked from my larger life jacket. Once passing the safety check station we were allowed to take out hot life jackets off and enjoy the views. The short boat ride was amazing. The ocean was a calm deep blue in the areas where you couldn’t see the bottom. The cliffs were sharp and stood proudly out of the water. Since the tide was low we could see that some of the rocks coming out of the water had been eaten away from the ocean and looked almost as if they were floating off the water from the indent at their base. The first beach we stopped at was packed full of people but still amazing. The tour guides told us 45 minutes at this beach. Some of the others went to play beach volleyball but I immediately headed over to the water with Julia to jump in. It has felt like forever since I got to dive under the salty sea. The ocean was warm but the further you dove the colder it got. The water was crystal clear and you could see a bountiful life under the surface. Chrishana joined us a couple minutes later followed by Jessica N. and Semaj. I kept diving to see what I could bring up from the bottom which seemed to be a lot. I found rocks and shells and so many pieces of coral that have been snapped off. After a while me and Jessica decided that we would try to launch each other out of the water to do some flips, although we were not that strong and they didn’t seem to work very well. After a couple more laps we heard the outpost boat calling for us and since we were a little late we had to run to the boat. Once we boarded we headed off to our next destination which was less than 10 minutes away. At this place we were told we could snorkel to see the amazing reef. I was so excited, I love snorkeling and swimming with the fish. After diving in, I struggled a bit to put my mask and snorkel on but after a few attempts I dove straight to start exploring. When I tell you it was beautiful, that is an understatement. The water was so clear and there was just so much life. I felt like I was in another world. There were Christmas tree worms in all sorts of colors, there were brightly colored clams who would get shy when you swam next to them. I found some anemones with some clown fish hiding in them and just so much more. The coral was so healthy and abundant, if you listened closely you could hear the parrot fish crunching on the corals which I loved. I said hello to a puffer fish who to me looked like the puppies of the sea. I even saw some spikey looking starfish! After diving a bit more and admiring the ancient corals it was time to leave, I could have honestly spent another hour or two there. The next place we stopped was a smaller beach where we had lunch. I was so hungry that I inhaled my warp in like 6 minutes. After the other finished eating we went off to the big Lagoon! This place is amazing, I sadly forgot to take photos but this place is known as one of the top places to visit in the entire world. We were put on kayaks and set off to start exploring. My kayak partner was Julia, of course, but we unfortunately only had one paddle which made it a bit difficult but we pulled through. The channel into the lagoon was spectacular, the sharp limestone walls protruded in some areas in which we had to skillfully watch out with our kayak (but it was difficult due to the lack of a paddle). The water was a blue I have never seen in nature before, it reminded me of the water cup you have when painting and you just dipped a blue paint brush into the water which previously had a white paintbrush in it. We tried our best to explore every nook and cranny of this lagoon, which turned out to be quite difficult. There was an area where the mangroves almost hid you and we decided to take a moment and sit there. The area was quiet but loud with bugs and the stiffness of no wind. The smell of the salty limestone was strong as we admired the plants dangling above us. Once both Julia and got our fill of the beauty we went to explore the next part. We paddled out to the middle of the large lake and decided to see where we would drift. It ended up taking us to the Jessica’s and they told us of a place where you could see the sea urchins. So we raced over to the back of the lagoon where there weren’t many people. Near an overhanging cliff an underwater shelf housed a dozen or so black pointy sea urchins. They were so cute! In a little pocket I decided to slip out of the kayak and take a quick dip. As I floated there just listening to the sounds of the lagoon I felt a small soft raindrop smack my nose. The rain was beautiful, it was soft and refreshing and made everything quiet. Then as we were heading back to the big open lagoon we headed to the outpost once again, meaning we had to head back to the boat. Oh! I almost forgot to mention that before kayaking we were doing some jumps off the boat where I thought I could do a front flip but the second one I ended up hitting my back. Once the kayaking was over we headed to our final destination which was another beach, however it was smaller and there were less people. When I dove into the water I felt something sting my face so I decided to stop swimming and rather play some volleyball with the others. Once we got tired of volleyball we all ran to the ocean to swim. We decided that we would try to throw each other out of the water and see who has the best flip. When it was my turn I felt like I was 7 feet in the air! It was so much fun. Once we got back to the hostel it was time for a nice hot shower and some rest. The next day we all got up pretty late due to being quite exhausted from the day before, and since it was a slow day I decided to talk to my family since it has been a while. The next day was similar as well, another slow start but then I got to go out on the town with some of the girls where we had some good food at a nearby restaurant then it was back to the hostel to pack for our departure. Check out was at 11 am and after we packed we waited for our which was scheduled to arrive at 1:30 pm. During that time I enjoyed a relaxing reading session on the beach with Julia. In the middle of it Julia did spot a very large hermit which we got to say hello to as we crawled off to his next place. The drive back was just as beautiful as the ride to El nido. Although it was long, it was quite pleasant after our fast paced weekend. Closer to the end of the drive coming around the bend there was a huge tour bus that seemed to have rolled over. Our diver stopped to talk to some of the others who seemed to be helping, he said everyone was out, and there didn’t seem to be many injured who were still on the crash site. We then had to hurry to the airport to catch our plane back to Manila. The flight was pleasant and I watched little women until we landed, and back to Los Banos it was. This was the best weekend I have had here, and will forever cherish the memories and the people we met there.

Until next time!

Lab work for the week

This week in the lab has been the busiest week I have had here so far. On Tuesday I started the day off by making media and Nutrient Agar slants. These are composed of…

• Beef Extract
• Peptone
• Agar
• dH2O

After briefly cooking the agar in the microwave the media is pipetted out into test tubes, each containing around 10 mls of the Nutrient Agar. Everything is then placed into the autoclave which takes around 2 hours to run. I have also been tasked with conducting other tests on the endophytes we have cultured. These include: Salt resistance, motility, Catalase, nitrogen levels, and phosphorous levels. Before proceeding with the experiments I needed to make and prep all plates and broths for the project. Jensen’s Medium was made for the nitrogen detection experiment. Pikovkaya’s plates were made for the Phosphorus detection experiment but two of the ingredients were lacking so we are not too sure if it will work. NA slants for culturing were made, and Tryptic soy broth tubes were also made for the Motility test. Once everything was taken out of the autoclave we placed the slant tubes on a rack that allows a slant to form in the tube. Once they cooled and solidified we were able to streak some of the colonies from the plates onto these slants. My supervisor says that slants are typically used for working cultures, I have never streaked on slants before so it was exciting. We put them in the incubator and waited till the next day to check on them. For the motility test a colony from the endophyte strain was poked into the center of the tube about half the way down, we are then observing its growth. Wednesday was a holiday so there was no lab work being done, but on Thursday is when I started to make solutions for my slat resistance test. I made 4 strengths of it: 0 mM, 50 mM, 100 mM, 150 mM, and 200 mM of NaCl. Along with the NaCl solutions I also had to remake my Jensen’s Medium plates because they unfortunately didn’t solidify overnight which was due to using too much Magnesium sulfate 7H2O, and Ferrous sulfate 7H2O. So we remade the agar and put everything in the autoclave. While that was running I ended conducting the Catalase test which detects an enzyme in the endophytes that catalyzes the release of oxygen. In order to prepare this experiment I needed slides and 3% hydrogen peroxide. I then smeared a colony from the strains and placed a single drop of hydrogen peroxide on it and waited to see if it bubbles. If it bubbles then it has the enzyme that catalyzes oxygen, and no bubbles meant no enzyme. Out of the five strains I tested only two didn’t bubble, those included: In-b-590, and In-b-24. The fact that 24 didn’t bubble is due to the lack of a sample. In-b-24 is very hard to grow and I may not have a large enough sample to see. Finally Friday came around and I was able to conduct my salt resistance test! I only had 10 Nutrient Agar plates left so what I had to do is split the plates into 5 pieces and marked the different concentrations of salt. Then I mixed the colonies in 500 microliters of the salt concentration and pipetted 50 microliters onto the plate. I made two plates of each strain and popped those right in the incubator at 37 degrees C. Then since my Jensen’s Medium plates finally worked I was able to streaked on those and place them in the incubator over the weekend. 

Weekend and Holiday activities!

As said above, Wednesday was a holiday so we didn’t have to go into work. So as a group we all headed to the town to the very small mall to pick up some stuff. I left my nice dress shoes at home so I ended up buying some $15 cute shoes for when we have to present them here! The others bought some clothes and a bag. After that Jessica N. has been telling us about this café that is super cute and super good so we went on a journey to find it. It was around a 10 minute walk from the main road, and we were pretty much walking through a cute little neighborhood. We finally stumbled upon and got ourselves into the cool AC building. The menu was great and I got a Spanish latte with cinnamon which was delicious. Once we were all done with our food and drinks we headed back. Typically we would have caught a bus back to IRRI but there were no more buses :(.

For the weekend on Saturday we decided that we will tackle the great Mount Makiling. The entirety of the hike starts at the base which you follow a manicured road up to the start of the hike to the peaks. But…. that would have added an extra 1 hour and a half to our journey so instead we paid 100 pesos to ride motorbikes up to the baes of the peaks! I have only ridden one motorbike before but this one was fun, although a bit scary especially since I felt us slip on a leaf. Once at the start of the trail there were some snack so I got a Turon for 10 pesos (it’s like banana lumpia but then coated in honey). Then we were off. I love hiking, it is the top most favorite of my hobbies. I hike a lot back home which is also a tropical rainforest environment, so this hike reminded me of home. There were many plants that seemed similar to those from home, for example they do have Lauhala (Pandanas) and there were so many ferns (if you know me ferns are my favorite). However there were also some plants that were super foreign, like trees which seemed to have some sort of parasitic fruit growing on them, or palms with thorns on them! There is also a large parasitic flower called Rafflesia. Sadly it wasn’t in bloom when we got to see it but they were still there. The one things that I did not appreciate on this hike were the leeches. We DO NOT have leeches on Kauai so when I got my first one of my thigh I freaked out. They were everywhere! Constantly biting, I was so happy that I have both long-sleeves and pants on. The hike was beautiful, maybe not the most spectacular scenery but the forest was stunning. If you sat quietly you could here the bugs humming and the bird singing, and if you listened close enough you could here some leaves clacking together similar to the olapa back at home. The guide who was with us would whistle a a very loud bird would respond, they seemed to be having a wonderful conversation! The dirt is similar to the mud in Pihea back at home, with clay in it which makes it more sticky and slippery. If it had rained this day we would have had an even harder time getting up. At the top of the peak which was peak two we could see the town but not much of it. The greenery is so thick and tall that we could only look at it through small clearings. Once we reached the rest point up top (station 30) there were butterflies dancing in the wind but then leeches crawling on the floor, I made the horrible decision to sit down up there and ended up getting a leech stuck to my butt (not fun at all). On the way down I also noticed how huge the tree ferns were! Overall the hike wasn’t super hard but I had a great time, it made me notice how much I miss home though. Once at the base of the mountain we got a jeepney back again and all took a much needed shower. I planned to sleep for the rest of the day, but no, because apparently two of the other interns wanted to come and visit Los Banos. I was tired, but still decided to go out. We ate at Seoul Kitchen where I got Sundubu jjigae which is a spicy tofu dish, it did have mussels in it so I gave it to Julia who has never had mussels! On Sunday our bodies were so sore that it ended up being a nice rest day. Hope you all have a wonderful week! A hui hou 🙂

Working in the Lab!

We once again had a long weekend with Monday being a holiday so my work week didn’t start until Tuesday. My supervisor had some errands to run so he was off for the entire day which meant no wet lab activities for me. However my day was full of researching literature instead! I was tasked with finding methods on extracting and measuring the components of exudates from roots. There was a lot of reading to be done but by the end of the day I had methods for bonds extraction and analysis. The methods to extract from soil typically may involve some suction and cultures of that soil in a nutrient broth. For roots you soak the root for 48 hours and then switch out the culture and soak for another 48 h. Exudates should be in the media after the 48 hours, all one has to do then is run the solution through a filtrate to only be collecting those exudates. Within the exudate extract there is a mixture of acids, bases and neutrals, in order to divided them one must use chromatography and columns. After my readings were complete the last task for the day was to water the rice seedlings for our experiment in the greenhouse. We won’t be using these plants as the main samples for our project they are for backup purposes only. on Wednesday is when we really started to do more things. Once my supervisor came into lab we made some media for our project. Typically there is a lab meeting on Wednesdays at 1pm but this week it was pushed back until Thursday. Right after lunch I was instructed to make 500 mls of Nutrient agar which consists of peptone, Beef extract, and agar. When that was completed I also had to make Nutrient broth which is the same as nutrient agar just without any agar added to the broth. once those were mixed up we placed them in the autoclave which typically runs for 2 hours. it was now around 2 pm by that time. While the autoclave was running, my supervisor, also the head of this project, took me to the greenhouse to transplant our seedlings of rice into bigger pots for growth. It was nice to get out of the lab, but those greenhouses are so hot. When I went to grab the last bit of plants I ended splattering mud up my shirt, it wasn’t too much and came off right when I washed it. Once we finished up our gardening for the week, we headed back to lab to see if the autoclave was complete. Unfortunately is was not which meant I would have to wait until tomorrow to pour my nutrient agar plates. Once Thursday hit we started right at 8am to get that agar from yesterday melted using a hot plate. the melting took around 30 minutes but then it was time to pour. Once I poured we realized that the agar wasn’t cooled down enough so it made some precipitation on the plates meaning i couldn’t streak endophytes until Friday. The reason we had to make Nutrient Agar plates is because four of the five endophytes that we chose from the freezer to be revived can only grow on Nutrient agar and not Kings B Medium. While we waited for the plates to cool we got in-b-24 endophytes out of the freezer and streaked it. These endophytes are stored in skim milk and glycerol so they had a sweet smell every time I opened the tube. once placing the plates in the incubator it was time to head to the lab meeting. I was instructed to crate a presentation about what I will be working on here in lab during my stay however, there were two other presenters and we didn’t finish the meeting until 3pm. Since my nutrient Agar plates were still drying we decided that we will streak the rest of the endophytes tomorrow morning. On Friday we started wet lab activities once again right at 8am. First we had to defrost the endophytes from the freezer which meant to leave them on ice for about 20-30 minutes. During that time I helped IA (my supervisor) with some more media he was preparing. Once the skim milk and endophytes were melted I vortexed them juts a bit to make sure it was mixed through all the was to ensure that I will be picking up endophytes and not just sugars. I streaked two plates per endophyte samples which cam out to a total of 8 plates. Once those were done it was finally lunch time. after lunch my supervisor had to run some errands again and I was told to find other biochemical tests I would like to conduct on the endophytes. Some that we are already doing are Nitrogen and Phosphorous levels. I read some articles and other test include Potassium levels, possibly drought and salinity tests and many more. I still have to run them by my supervisor to see exactly what we will be testing. I look forward to isolating more endophytes next week.

Weekend Activites!

After completing all my work I was able to get off of work an hour early which was helpful because the we were going to Manila for the weekend and our bus was picking us up at 5:30 pm. We finally got to Manila around 8pm and checked into our hostel. I have only stayed in one hostel before and this one was quite nice compared to the other one. We booked a 6 person room which meant that we were the only people in that room. The next day we got up around 9 and didn’t head out until 11am, we decided to head down to the café right below our hostel which was called Sasa Café. it was cute and the food and drinks were delicious! After wards we headed out to explore some of the museums. Sadly however the only museum we got to explore was the anthropology museum, I noticed that the group with us (Jessica, Julia, Jessica) very much enjoy Museums and wanted to make sure we saw everything. After about many hours in that Museum we wanted to head to the Mall Of Asia Also known as MOA to meet up with some of the others from FNRI. We spent many more hours in the mall where I bought some skin care products and of course food. After our shopping we were all so hungry so we had dinner at a cute ramen shop whcih had some very delicious ramen. Later we headed back to our hostel. Sunday was our last day in manila and the grils went to church but I am not one who goes to church often so instead i went to the mall to get some food with Sam. after spending a little while there we headed back to our hostel to check out and get picked up by our bus to head back to IRRI. And that was the weekend!

The second week started off to a slow start. I began the week in lab with reading some more materials relating to the project we will be starting soon. The project is surrounding the main idea of reducing methane production in rice farming by finding a more friendly long term solution. I began this project by making some media and plates, these included LB broth and plates, Kings Medium plates, PDA plates, some sterile water and 50% glycerol. All these materials made are in preparation for the isolation of microorganisms from the leaf and roots form the rice varieties we have. Later that day on Wednesday I sat in on a lab meeting where the advisor I am working with presented the proposal of this new project to the entirety of the lab. It was good for me to get a better in depth view of the overall project. While presenting the rest of the lab would put in some comments on what would make the project go more smoothly and other advise. For example we will be using rice samples from the long term rice experiment in order to have samples that have exposed to non-controlled rice samples. We may also use some endophytes that are stored in the lab to start with our experiments. I had a short week last week with my day ending on Thursday where my advisor wasn’t in lab so I just ended up ding some computer work that day. But then on Friday we got a tour of IRRI starting with the Library and ending with the gene bank! The gene bank was so cold that we had to wear some heavier jackets but it was deficiently an amazing room to see. The tour allowed us to see the other facilities that IRRI has to offer. We visited the seed health department in which they make sure the seeds are healthy, we also visited the growth facilities where they have growing rooms which enable the perfect amount of sunlight, humidity, and carbon dioxide levels in the room. Lastly we had a tour of their farm. Which we got to see the larger scale experiments being put to the test in the large rice patches.

Finally the weekend hit and we decided as a large group with more of the interns to go to Villa Escudero. 12 of us went there for the whole day, We had to wake up to leave at 7 AM in the morning, and f=drove around an hour to Tiaong where the resort was. We began by visiting the museum which had many artifacts explaining the religion then on the top part of the museum it had more displays of wildlife found around the world as well as some history on the Philippines. Then we got to ride a water buffalo carriage to the resort area where there was a pool and a rafting pond. We decided that we would do the rafting which was so much fun. It was a two person raft and my partner was Julia. We had such a great time. However, it was very hot so after rafting we jumped in the pool till lunch which was at one. Lunch was at the base of a man made waterfall where there was traditional Filipino food served buffet style. We spent a while admiring the waterfall and getting our full of the delicious food. To end the day we spent it once again lounging at the pool.

Overall the Philippines have been welcoming and a great experience. I think I am finally getting used to the humidity here. I look forward to the work a head me! See you soon 🙂