Still too much growth

Grrrr….once again I am running into the same old problem with way too much growth on my plates. After trying and trying again with different amounts of yogurt or milk being added into the peptone water I still cannot get countable CFUs. The first dilution used to plate was a solution of 99ml peptone water with 1ml of yogurt or milk. Then I used that solution to plate with different amounts (100µl, 75µl, 50µl, 25µl, & 10µl) on five different plates for each yogurt or milk. When the first set of plates came back with too much growth I moved onto trying again with a different amount of product. The second dilution used to plate was a solution of 99ml peptone water with ½ of 1ml of yogurt or milk. Again the solution was used in different amounts on five different plates. Still I kept getting too much growth so I went even smaller with the amount of yogurt or milk being added into the dilution. I gave it two more attempts with the amounts of 1/10 of 1ml and 100µl of yogurt or milk; believe it or not there was still too much growth.

So after realizing that this course of action was not working I went to Cori for help. She suggested that I try to dilute my dilution solution instead of changing the amount of yogurt or milk being added into the peptone water. So my new course of action is to start with 99ml peptone water with 1ml of yogurt or milk then I’ll be taking 1ml of that solution and adding it into a new peptone water then adding 1ml of that solution into another new peptone water. I’ll keep repeating this process until I reach the fifth peptone water. After I have the five dilutions I’ll be taking 100µl from each dilution and plating it. I am keeping my fingers crossed that this course of action gets me the results I need to move forward in my project.

The top picture is of the five dilution of peptone water I prepared inside the lab to see if I can get countable CFUs. The bottom picture is from http://www.personal.psu.edu/ to show a more clear understanding of what I am trying to do to get countable CFUs

Redo week...

I don’t have too much to report this week in lab. Aside from having to re-plate because the plates were left in the incubator too long, I haven’t moved too far ahead in my project this week. I am trying to get some good/countable CFU (colony forming units) so I can compare results after putting the yogurts, buttermilk, and kefir milk through the mockup of the human stomach I’ll be making inside the lab. Speaking of the human stomach mockup I had a difficult time coming up with how to actually do something close to the stomach. After brainstorming about it and talking with Matt and Josh I finally came up with something I can do with the resource within the lab.

I’ll be using a recipe of hydrochloric acid, sodium chloride, potassium chloride, and distilled water to make up gastric acid of the stomach. After making the solution a proper amount of yogurt/milk will be added to it and then placed inside a Ziploc bag. Once inside the Ziploc bag it will then be placed flat inside a box with marbles laying on top of the bag. The whole box will then be put onto the rocking machine inside the lab so the marbles can move back and forth atop of the bag. This is what I have come up with to simulate the muscles of the stomach. I’ll be running the whole stimulation for two hours on each of the products because it usually take about two hours to digest food. I am looking forward to actually running this stimulation when the time comes. Well fellow S-STEM scholars until next time and good luck with your projects.

Picture of the redos done this week there were sixteen done in all. Four for each product with different amounts on each one. The amounts were 25µl, 50µl, 75µl, and 100µl. Using a 99ml peptone solution with 1/10 of 1ml yogurt/milk product.

Off to a good start...

This week in lab I started plating my two yogurts and kefir drink using the L. S. Differential media I made last week. The first set of test I did on Monday came back with too much growth on the plates. So I had to readjust the amount I was using on the plates. I first start out using 100 µl of solution on the plates but that produced way too much so I lowered the amount to 25 µl. The second set of test plates were done yesterday (Wednesday). It takes about 48 hours in the incubation process for growth to happen. I’ll be checking on the plates tomorrow (Friday).

Other than plating this week in lab I have be gathering material together to make up my human stomach simulation. After checking with Cori to make sure that the lab had all the ingredients I needed to make up my gastric acid. I got the green light that the lab has all the ingredients I needed. I’ll be using a recipe of hydrochloric acid, sodium chloride, potassium chloride, and distilled water to make up gastric acid of the stomach. I have also been working with Josh to come up with a way to react how the human stomach digest food. We came up with a pretty good way to do it but I’ll share that next blog. Got to have something to write about next week. Hope everyone’s projects are coming along nicely and I hope you all have a great weekend.

The pictures above are of the materials I used to plate this week and of the first set of test plates done on Monday October the 20th, 2014.

Making Media

Top picture is of L. S. Differential medium base & the bottom picture is of the antibiotic-free skim milk.

Yes after waiting and doing some more waiting I actually got to physically start my project this week in lab. All my supplies finally came in to make the L. S. Differential media I’ll be using in my project. On Monday I talked to Cori about how long it would take from start to finish. She told me that it would take about three and half hours. It was a no go on Monday for making the media because I wouldn’t have enough time to finish.

On Wednesday I came into the lab and made the media. I’m not going to lie it took me more than three and half hours to make the media. However, it was a great learning experience. I ran into a few problems making the media but nothing I couldn’t overcome with the help of Cori. The first problem that came up had to do with adjusting the timer on the autoclave machine. The L. S. Differential media requires antibiotic-free skim milk to be added into the main media along with TTC (Triphenyl-Tetrazolium Chloride). In preparation the antibiotic-free skim milk had to be autoclave for five minutes and the setting on the autoclave are set at fifteen minutes. When we asked Josh about how to go about changing the settings he brought up another way to prepare the skim milk instead of changing the settings. That way was to use the microwave to heat up the milk and then filter it through a filter pump. This way did not work at all and caused most of the problems that I ran into. From the microwave over cooking the milk to the filter getting clogged do to the fat cells inside the milk. In the end Cori found a way to readjust the settings using the liquid settings on the autoclave machine and I ended up autoclaving the antibiotic-free skim milk.

The Waiting Game

After doing some research last Friday I came upon two different test solutions that would simulate gastric fluid and intestinal fluid. I took my research to Matt to make sure it was doable inside the lab and it turns out it is. Matt had to order a few thing to make it possible to make the fluids inside the lab which should be coming in sometime next week. So as of right now I am just waiting around for the items to come in but I have been using this time to type out my abstract, background information and procedures. It’s never too early to start thinking about the rough draft paper that will be due next month as we get closer to the end of the semester.

Image is from http://www.firstcovers.com

The digestion process - What happens to your food as it travels through ...

This week in lab I worked on getting the knots out of my project that keep me from getting it off the ground. After brainstorming for what seem like forever I came up with a plan that I’m happy with and most important I got the okay on. I’m going to be testing the effect of stress that probiotics go through inside the digestive system to see the survival rate. The two probiotics that I’ve chosen for this project will be Lactobacillus bulgaricus and Streptococcus thermopilus these two probiotics are mostly found in yogurt… Yep that’s right I’ll be using yogurt once again however this time around I’ll be only using three yogurt products instead of twenty products.

So the first thing I’ll be doing is testing each product to make sure they contain the probiotics I’m looking for. For test one I want to use LS differential agar but we don’t have in the lab. I guess I’ll have to talk with Matt to see if we can order it. This media would be able to differentiate between Lactobacillus bulgaricus and Streptococcus thermopilus. In the mean time I’ve been talking with Cory to see what else I could use that we already have inside the lab. After I get done with test one I’ll be moving on to the stress part of the project. Taking the probiotic and putting it into an environment that is like the human stomach. It looks like I’ll be use hydrochloric acid because it’s very close to the pH level of the stomach. I’m still thinking of what other material I’ll be using to create a similar environment of the stomach.

Video from McGraw Hill on youtube.

Breaking out the anatomy textbook

This week I have been researching how the digestive system works and mapping out the probiotic project. After talk to both Matt and Josh I got an understanding of where I want to go with this project and how I can accomplish it. The base of my project will be how different products with probiotics hold up in the digestive system. I will be measuring to see which product will have the most lactobacillus produced.

My first step was doing research on how food get digested in the stomach and moves on to the small intestine. While researching I also found out the pH levels of the stomach and small intestine. By getting this information I am trying to come up with a media that will be similar to the gastric acid in the stomach. There are other items that still need to be mapped out but I am happy to say that the probiotic project is coming together.

When it comes to the products I’ll be using I have narrowed it down to five products. I was going to use a pill supplement but after talking to Josh about it I decided to can that idea. There are just too many supplements out on the market. Instead I am going to stay with buttermilk and yogurts (three different types - homemade, Greek yogurt, plain yogurt). I will also be adding kefir into the mix. Kefir is a fermented milk drink made with kefir grains and has its origins in the north Caucasus Mountains. It is prepared by inoculating cow, goat, or sheep milk with kefir grains.

Image from http://www.aspirus.org

Continuing with probiotics project but I get to do the straw gardening as a side project too.

Hello everybody,

What a crazy start to this week with all the rain on Monday. I hope the rain didn’t cause too much trouble for anyone. As my title states I’ll be continuing my work with probiotics but this time around I’ll be taking products with probiotics (not just yogurt) and comparing them to one another on how they affect the digestive system. I am still working on getting this new project off the ground so I don’t have a lot to go on right now. I’ll be spending the next week doing research on the digestive system and see how I can bring this project into the lab.

As I stated last blog I really wanted to do a project with gardening. With the help of Paul I found a project that was totally cool to do. I ran it pass Josh and Matt and got the green light to go but ran into problems off the bat. Problems like where would we put the straw bale at??? Another problem that came up was the maintenance of getting the straw bale project started. Josh brought up the idea of doing the straw bale project on a mini scale so it would be easier to do inside the lab but I would really like to do the real thing. So as of right now Josh and Matt has said that I can do the straw bale project but we are still working out the problems we are facing.
Image from http://www.liquidplanner.com

A START TO A NEW SEMESTER / A START TO A NEW PROJECT......HOPEFULLY SOMETHING WITH GARDENING

Hello everyone, I hope everyone had a great summer. It’s a new start to a new semester and that means a new start with the S-STEM program. I am happy to be back and ready to start working. I am unsure of what project I’ll be doing, I still need to talk with Matt and Josh about it. However, I’m thinking I want to do something with gardening along the lines of growing something. Over the summer I got into gardening with my nine year old niece Clarice Marie. Our family garden was not the best success but we did have a lot of fun trying. The best part of it all was spending time with my niece and getting our hands dirty. Another reason why I want to do something with gardening is because gardening has been my go to thing to relax. By the end of last semester I was stress and it didn’t help that family issues arose at the same time school was finishing up. I needed to find something that would help release my stress and gardening was it. The way I look at it is why not have a project I can work on and at the same time relieve stress. I look forward to working in the lab once again this semester and seeing everyone. Image from http://www.exploratorium.edu/gardening/

Hi all, I don’t have too much to report this week. I only made it into the lab two days this week Monday and today (Thursday). I had a family emergency come up early Tuesday morning that kept me from coming into the lab on Tuesday and Wednesday. Besides working on the research rough draft I collected all my data from my notebook so far and made a excel spreadsheet. Below is just a glimpse of the finished spreadsheet. I couldn’t fit the whole spreadsheet into the print screen but the final spreadsheet has all twenty yogurt/dairy products on it and the tests done. The count of the tests done so far is eight however not all the tests were done to each product.

One thing after another……..feeling the pressure.

Last week was a very busy week for me. I had two test one in biology and the other in chemistry. So I did not post a blog or do the discussion bored. To top it off I had to start out this week with a test in math…..grrrrr! As the week went by one thing after another kept popping up adding more items that I need to take care of like the reapplication for the S-STEM program for fall 2014, the rough draft for the research paper, and let’s not forget the conferences at Metro Tech and Estrella Mountain. Speaking about the conferences I am starting to get a little worried because I am not getting the results I need to wrap up my research. I am either getting no growth or too much growth I cannot count. I hoping that I can get a count on all the yogurts by the end of next week fingers cross. I will be heading into the lab tomorrow to make a whole bunch of TSA plates so I can finish getting the results I need so I can move on to the next step.

Picture from bentbutnotbroken.org

Brand new yogurts yea!!!

Hello all, I hope everyone had a wonderful relaxing spring break. I know the last time I blogged before spring break I said I had a game plan which I did. However once I got into the lab on Monday I was stopped in my tracks due to all my yogurts and dairy products being out of date (expired). I had to make a shopping list of all eleven dairy products for Matt. It took a few days but Matt came through with a whole bunch of yogurts and dairy products. To be precise he came through with eighteen yogurts/dairy products; the refrigerator is full to the max. I have to wait until next week to dive into all these new products I cannot wait to start. With that many products I will be dividing them up into five groups; below are pictures of the groups I have divided them up in.

Time to take a break and recharge

I don’t know about all of you but I am very glad that next week is spring break. I plan on spending my getting caught up/head on homework, getting caught up on house hold chores I have been putting off and taking the time to really relax so I can come back recharge ready to hit the ground running. Since next week I won’t be in the lab I took time this week to wrap up what I was working on which was trying to get a count of growth from the yogurts I am working with. Out of the eleven products of yogurt I’m working with I only successfully got four of the products to where I can actually count the number of growth. Everything else has come back either blank, contaminated, or too numerous to count. I cleaned out the incubator of all my trails and error plate and made more TSA media so when I come back I can jump right into what I need to do. I also came up with a list of what I have to work on still and with any luck I can get most of that list finish the week we come back from spring break. Well I hope you all have a great and safe spring break see you all when we get back.

Image from wordfromthewell.com

Writing Time

So this week has been spent playing catch up in both my classes and in the bioscience lab. I don’t have much to report this week aside from working on my abstract for the student conference at Estrella Mountain; I redid four of the yogurts because when I did them in the first place I mixed up the amount of yogurt that was supposed to go into the solution. With getting over being sick I didn’t make it to the field trip last Friday to the World Wildlife Zoo. I was really bummed I couldn’t make it but I was right in my decision to stay home and rest up. I look forward to seeing everyone’s blog that got to go and took pictures. Have a good weekend all!!!

This image was taken from grammar.about.com

Sour throat, runny nose, and cough oh my........

Howdy all, well today is my first day back in the lab this week. I was unable to come in because I caught a cold bug over this past weekend. I woke-up Saturday morning with a sore throat and a mild cough, by Sunday morning I could add a runny nose to the list of symptoms I had. I knew I had to take some time to rest when I couldn’t stay asleep Sunday night because I kept on coughing. Waking-up Monday was the worst; Tuesday was a little better but I decided I better not chance it and go to school. I am feeling better than I did this past weekend but I still have a cough that is still lingering. Upon arriving in the lab today I found out that I made a mistake while diluting some of the yogurts last week. I changed the peptone water to 90mL from 99mL with making this changes I over looked changing the yogurt amount from 1mL to 10mL (99mL of peptone water with 1mL of yogurt and 90mL of peptone water with 10mL of yogurt). This is something I’ll be mixing next week. Since I only made it in one day this week I wasn’t able to make another batch of TSA media, so I don’t have any pictures to post. However, I did take a picture of the already finished TSA plates I did last week (posted blow).

A week of studying and trying something new.

Howdy all!! Who else is ready for the three day weekend coming up? I know I am, this week has been packed with studying for tests in both BIO 182 and CHM130. I also finished up diluting the rest of the yogurt products I had not gotten to last week. The pretty cool thing I did this week in lab was learned how to make TSA plates. I was using so much in my project with the yogurt Josh had me make my only plates. I totally forgot to take pictures when I was doing it, however I am going to be make another batch of plates come this Tuesday so I will make sure to take plenty of pictures. Have a great weekend everyone.

The above picture is from amazon.com

Moving Forward

Hello everyone, hope you all had a great week so far. Well as my title states, I moved forward in my project to diluting the yogurt before putting it on a TSA plate. With eleven products to dilute and plate I decided to split the products into three group; two groups of four and one group of three. I diluted and plated the first group of four products on Tuesday the 4th and the second group of three products on Wednesday the 5th. Out of the first group, only one plate had growth on it (I am looking for Lactobacillus or Streptococcus) the other three had nothing. From the second group the same thing happened only one plate had growth and the other two had nothing. The two plates that had growth were the Fry’s buttermilk and the Mountain High plain regular yogurt. However, the two plates had too much growth to count so I will be re-doing them next week with a lower amount on the plates. As for the plates that had nothing I will also re-doing them but this time I will be changing the dilution to a lower solution. Next week will be a busy week full of re-dos and finishing diluting and plating the products I did not get to this week.

The two top pictures are of all the equipment I used to dilute and plate. The bottom two pictures are of the plates that had growth. Second to last picture is the Fry's buttermilk and the last picture is the Mountain High plain regular yogurt.

NEW SEMESTER, NEW PROJECT.

Welcome back everyone and a big welcome to all the new S-STEM scholars. I am happy to be back and ready to make this semester a great one. This pass Monday the 27th was my first day back in the bioscience lab and there was a change made to my project for this semester. Instead of fomites being my project I’ll be doing a project on probiotics. The new project seems like it going to be fun to do; I’ll be working with different types of yogurts/dairy products. When I came in on Monday Matt had already gone to the store and picked up eleven different types of products to be tested. Matt gave me a good starting point to start at; he hinted at looking up how to detect concentration levels in yogurts and dairy products. After looking up what I needed I decided that I would start with streaking a TSA plate for each eleven products to see if I can grow any bacteria, I did that on Tuesday. When checked today some of the TSA plates did have some growth on them. However, after talking to Matt he said that I started at a good test but what would work even better is if the products were diluted before putting them on the TSA plates. So that is what I’ll be doing next week.

Above are two pictures out of the eleven TSA plates streaked.