Week 2: Initial Test and Water Change!

 Hello! Hanah here again✌😁

So today IS THE DAY of the first water change. I prepared the new conditioned water with all its treatments and plant nutrition. I sealed it in a 5gal bucket with Saran Wrap and tape to make sure nothing got in until I can buy a proper bucket with a lid from Home Depot or something! I couldn't change it yesterday since I need the water to be properly conditioned over 24hrs. I also began to label each syringe I would be using for each chemical and plant nutrition fluids and put those instruments in separate baggies, one for Tank Water chemicals and another for Plant nutrition chemicals.

However, I did go ahead and my first water test with the Fresh Water kit and recorded my initial data for the tank this week. Everything was good except the pH and the Nitrates. I was a little annoyed since this has been the issue since the passing of my other snail in the tank/: So, no. Not that good. Here is a picture of that.

Even though I'd be more alarmed if the Ammonia and Nitrites were high instead of Nitrates, this is still something I need to fix soon before Vincent(betta fish) and Gary(Nerite snail) begin to get affected. To my fortune, this is easily fixed with good and big water change. I decided to do an initial 60% water change to not stress the fish with a drastic change tonight. Every week, I will be checking the water for Nitrates and doing 20% water changes as needed until I can get the Nitrates level back to 0 ppm. Something awesome that I came to learn is that if the Nitrate levels increase, the pH levels decrease significantly. Realizing this honestly made me feel a little more confident of a good research question and a much better hypothesis revolving around these two variables. I'd love suggestions if you have 'em! Some of my constants will be temperature and volume, and waste(food digested by Vincent and Gary). I will also be controlling the amount of time I will be leaving the light on in the tank since I am currently having a big algae problem.

Vincent has been digging some of my front plants, so I will be replanting those. During the water change, I will be taking Vincent out into a separate small tank, including Gary,  to gently scrub the algae off of the plants and his house. I want to do some trimming to the dead leaves since that can also influence Nitrate spikes. I will also be doing a 50% change in the biofilter media just enough so there isn't too much sludge but enough so that the beneficial bacteria aren't disturbed. This IS the one thing that helps break down ammonia and nitrites and keep the cycle going, so we do NOT want to kill that.

I will be doing a water test again after 24 hours to see how things are, then check in a week again before the next water change.

Week 1 Spring2021!

Hello Everyone!

I'm super excited to finally be working on TRAIN stuff once more! I'm certain it has a lot, if not everything to do with the new research project that I jump-started a couple months back in the midst of COVID-19. I'm talking about my 5-gallon live plant aquarium which is currently housing a male red betta fish called Vincent and a snail called Gary. The was a smaller snail named Beetlejuice but he sadly passed unexpectantly, I'm sure he had already been a bit since when I got him. I'm hoping to be adding a second snail to give Gary some company soon, though. The main question for my research is whether or not I can create a balanced ecosystem in my fishes tank with the help of beneficial bacteria, live plants, light, in a 5gallon tank, and only having to worry about minimal water changes and monitoring the pH, temperature, ammonia levels, nitrites, and nitrates. so far I have successfully maintained the tank going for 5 months, and I've only had to change 25% of the water maybe every 3-4 weeks.

    

This next week, However, I will have to do a big water change, and also replace some of the biofilter media due to the sudden passing of one of my snails. I first noticed something off when I got a very high reading of nitrates in my tank which is harmful to the fish, and that's when I realized it was due to the decomposition of the dead snail.

I also got a bad case of green and black algae on my plants, so I did have to trim down some leaves to stop the decaying of some plants thanks to the ALGEA. 🍃

SO. It feels a bit like a downer, but this is also perfect because I get to give my tank a little restart just in time for TRAIN! I'll do the water change and filter stuff next coming week and will keep you all posted! Good luck and Happy Valentine's day!💖

WEEK 7 Fall2020

Hello Everyone, hope your week is off to a good start!

I was gone for a while thanks to life(this whole year really), but thanks to a small chat with Dr. Chapman I was able to get back on track with everything. I've decided to put my current research project on pause for next semester just because I feel like I'd need the complete semester for it, due to the fact that its all a combination of Bacterial biochem shtuff and coding, which I have no clue and where to start with the last part. I do have an Excell document started which has been serving as my "database" for now. However, due to not really being able to be in the lab, things got a little more complicated for me. I am a person that likes getting things done good and give 100% of my best effort. I don't like doing nor leaving things halfway. So with Dr. Chapman's advice, I will be putting a pause and start that research official in Spring.  In the meantime, I will be participating in a few tasks Dr. Chapman has set out for me that will still keep me busy and learning.

Now, on to Halloweenie things... My betta fish disappeared completely. Yes, I checked around and under the surrounding furniture, including filters and whatnot... I CAN'
T FIND A BODY.

Two nerite snails visible.
 1 next to the heater bar, and the other under the big leaf

I was adding a new live plant to its aquarium and a new nerite snail to the tank; then after cleaning everything up, I fed him, Leonard (my fish), closed the lid and went to get lunch. I come back with my dad to show off the new set up... and HE WAS GONE.  3 days later and he is still gone. What is weirder is that I only have guinea pigs aside from Leonard... which unless they pulled a G-Force stunt on me... and became pescatarian...Are far from being suspicious. I mean look at them.

The mystery is what mostly bothers me at this point, Like not being able to even find the body??? No water signs anywhere?!

Anywho. I hope your week goes smoothly and I wish you the best on your midterms this week! I look forward to catching up on what the rest of you have been up to. Let's hope and pray the spooky things stay at bay!

 

WEEK 5: More Gram Staining and Frozen Plate??

         

On this week I went ahead and streaked a Hektoen Enteric agar plate with a colony from the isolation plate of Salmonella enterica just to make sure that this plate in fact had ONLY Salmonella growing on it, since the colony growth on it was unfamiliar. The HE plate is a selective and differential agar primarily used to identify Salmonella from an unknown culture.


To my surprise, the HE plate showed positive results, meaning that this colony is Salmonella. However, the results also indicated that this microbe didn't show indications of lactose fermentation or hydrogen sulfide production; so it remained its original green color.



I then went ahead and made Gram stains for B. subtilis, S. aureus, E. faecalis  to take a look at the cell morphology of them. Unfortunately, I smeared too much bacteria for B. subtilis so could not really see much of the cell shape due to how much clumps were present on the slide.

       

WEEK 3: Cross Contamination & Gram Staining

On February 13, I went ahead and began another isolation plate for E. coli due to the amount of time that had passed by. As you can see in the images below, the original cocci or circular shape of E. coli cells had denatured into coccobacilli mostly because of how old the bacteria had gotten. In addition to the E. coli isolation plate, I began one for Bacillus subtilis.  Both isolation plates were placed in an incubator to stay overnight and be checked for growth the next day.

The next day on February 14th, as I observed the growth on the TSA Isolation plates for E. coli and B. subtilis, I discovered that the B. subtilis plate had been contaminated with a different type of microbe. With further observation, we came to the conclusion that the broth culture of B. subtilis had been contaminated considering how much precautions I had taken to inoculate the first isolation plate. Therefore, the culture that I used to inoculate was disposed of, and a new culture was used to inoculate a new TSA plate to isolate some colonies of B. subtilis. 
After this was taken care of, I proceeded to move on to observing the E. coli plate, and after seeing that it was good, I went ahead and did a Gram stain. this time the cell shape was circular and violet.

WEEK 4: The Start of the Bacterial DATABASE

Hello everyone! I hope everyone had a lovely long weekend, I certainly took the opportunity to catch up on sleep (finally). This week in the lab was a little exciting, for the conversation of the database for the bacteria research began and plans began to be made. The first thing that was made, was a list of all the possible tests that can be conducted in our lab to identify different characteristics of the bacteria that we will be looking at. In total there were seventeen tests, plus cell and colony morphology observations that can be conducted in our lab for each bacteria; which I am more than excited to be conducting. 

With the help of Matt, we both started a Google Sheet and began a database where the data gathered from these tests will be inputted, and will be shared between myself, Brittany, Matt, and Josh for now, since we are the main ones on this research project. It definitely took me a while to find an appropriate chronological order for these test to be done, but I have to give a huge shout out to Daisy for sharing her advice and experience with me to sort these test in a good order.  Once this step was done, the next task was to come up with a system on how we were going to input data. So we did a second sheet tab as a key, and we used this tab to create a drop-down for each Test column using the data validation tool. I this was probably the coolest part of this whole creation. At least for ME because I didn't have any experience with Google Sheets or Excel, so I can't deny the fact I felt kinda cool knowing this trick 😆.

The database still needs a lot of work of course, but what we have right now is a pretty strong base to build from. 

Week 2: ROOKIE MISTAKES

For week two, I was able to finally check for growth on my Petri dishes that I had previously inoculated with the bacteria commonly known as E. coli. 
I had gotten good results at first, but to my misfortune, I left the plates too long in the incubator and ended up drying up the media.
I pretty much should've known better, but I guess this is one way to learn.
That led me to finally take the correct precautions and do another set, which ended up giving me very lovely isolated colonies.

I decided to wait out the next procedure, which is to gram stain the bacteria so that I could identify if E. coli was a Gram-negative or a Gram-positive bacteria, and also determine the type of morphology it had. This is usually is determined by first, identifying under a microscope whether the bacteria is in pink color or a dark purple. Then we observe and determine if the bacteria has a rod, sphere, or spiral shape that will eventually help us determine the name for this bacteria.
I hope to get good, detailed pictures of this so that I can have a nice reference and visual image I can look back to. 

Below are some pictures of one of the Petri dishes I completely ruined, and one that I did correctly.