12/7-12/8 The Making of Cheese (Part II)

Wednesday (12/7) Ilde showed Tomie and I the 3 quadrant inoculation technique, which Tomie posted a picture of. Using this technique, we took two new samples from each cheese and put them in the incubator. Then, we gram stained bacteria cultures from last week's plates (FYI: gram staining is a method for staining samples of bacteria that differentiates between the two main types of bacterial cell wall). We started off by placing 1-3 drops of DI water on a microscope slide and mixing a small amount of bacteria to it (using an inoculation loop), we did this for eight cultures. We waited for the slides to air-dry, then began the actual staining process...

Stain/dye with Gram Crystal Violet for 30 seconds

Rince with distilled water for 5 seconds

Use trapping agent, Gram Iodine for 60 seconds

Rinse with distilled water for 5 seconds

Use Gram Decolorizer for 1-5 seconds

Rinse with distilled water for 5 seconds

Stain/Dye with Gram Safranin for 30 seconds

Rinse with distilled water for 5 seconds


(Note: Only enough drops to cover the dried bacteria is needed)

We didn't get to slide eight because we left to meet with Mrs. Chapman to discuss our Projects for next semester and our experience at PC thus far.


Thursday (12/8) Tomie, Ilde and I worked with misroscopes to identify each of the bacteria. We checked if each one was Gram Positive or Gram Negative, then evaluated the shapes to determine the type of bacteria i.e. Spirilla, Cocci, Bascillus. We were successful in identifying most as either Staph or other type of Bascillus (I can't consult my notebook, which is at PC this very moment missing me). But there were some we couldn't i.d., even after consulting Rick and holding him captive for over an hour, and we weren't comfortable labeling them.

11/30-12/1 The Making of Cheese (Part I)

Two weeks ago Tomie and I started two small projects to try to make cheese...

Wednesday (11/23) we mixed a tablet of rennet into 400mL of DI water, then added 40 mL of it to four different beakers containing: a 400mL milk/buttermilk solution, a 400 mL milk/yoghurt solution, and two filled with 400mL of milk. We each had a milk/rennet solution but Tomie was responsible for the milk/yoghurt/rennet solution and I the milk/buttermilk/rennet solution. After stirring and covering the solutions we put all of them except the one with buttermilk in an incubator at 37 degrees celsius until the next day. The one with buttermilk was left at room temperature. (Note: We missed a step where we were supposed to heat and then cool the milk before adding yoghurt, buttermilk or rennet.)

We arrived Thursday (12/1) and drained the excess liquid from the solutions using a filter. The milk/yoghurt/rennet solution had pretty solid cube-shaped curds, the milk/rennet solution Tomie was responsible for looked like cottage cheese, while the one I was responsible for resembled cottage cheese but wasn't quite there, and the milk/buttermilk/rennet solution had a liquidy-yogurt consistency. The last thing we did was add Kosher Salt. (Pictures in written order, left to right)

The pH levels of the products ranged from 6.0-6.9. We took two samples from each beaker, plated and incubated them.

12/7/11

Yesterday was a pretty slow day, well its usually a slow day on Wednesday because most of the classes don't have any labs but this particular day was the slowest yet. There wasn't that much to do on this day because Josh told us that we needed to meet with Amanda to discuss what we have done in the lab so far, Basically everything we have done from the moment we got to Phoenix College by going of the blog post we have made. We looked at the cheese that we made and it didn't look like cheese, more like some oddly deformed piece of yogurt substance which was really disgusting because I really dislike yogurt with a passion the smell of it just makes me want to vomit. After we checked the cheese we made Matt told us to inoculate some of the cheese in a TSA medium  which is basically a jell like substance that makes bacteria grow we also learned how to do a 3 zone inoculation and it was fairly easy basically its just dividing the dish into 3 parts and then streaking it from one zone to the other but after you do the first zone you don't go back to it.

12.07.11

So yesterday was really slow. Mainly because we were waiting to speak with Amanda. What we did was look at the cheese we created. We forgot to lable which one was made with buttermilk and which one was made with yogurt. So Mat asks us to inoculate the cheese to see what bacteria was used. The one, we believed, was made with buttermilk seemed more like yogurt than it did cheese. The other one was starting to really resemble chesse, so that was pretty cool ( cause that was the one I made) =)

We also, kind of, learned how to do the 3 zone inoculation. I say kind of because our teacher, Ilde, showed us but did it wrong himself while doing it. But its ok because we still understood it in the end. Last, but not least, we all sat and learned about the rubric for out projects. We also got a chance to tell her about our expierences here and what we've done. Pretty cool. =0

This is the picture of the 3 quadrant technique from zone 1-3.

12/7/11

Today we were gram staining our plates we had made from our cheese and had made new plates using the 3 quadrant zone. When gram staining, it's pretty much almost vital that you get the times correct or else it will destroy your data and alter your results. They way we gram stained was using this substance called Crystal Violet for 30 seconds. 2nd, we washed it off using distilled water for 5 seconds. 3rd, we put on Gram Iodine for 60 seconds. Then washed again with the distilled water. Mind you that we used the little droppers in the bottles it came with so we did a few drops to cover the bacteria. 5th, we put on the Gram Decolorizer for 1-5 seconds. 6th, washed off with distilled water once more. 7th, put on Safranin for 30 seconds and washed it off, yet again, with distilled water.

I learned a new technique of inoculating bacteria and that's the 3 quadrant zone which we spread the cheese in half of the plate, sterilized the inoculation loop, spread the cheese in a quarter of the plate, sterilized the inoculation loop again, then spread the cheese once more over the last quarter. We then taped up the edges and stored it in a "special fridge".

Lastly, we meet with Mrs. Chapman to talk about our upcoming projects and what we have been doing thus far in the semester. :)

11/30-12/1/11 SAY CHEESE!

CHEESE!!!!! Making cheese might seem hard and it might seem easy, depends on the person. Having the Rennet tablet dissolved (rennet is an enzyme in the baby calf, sheep, goat that breaks down the mother's milk), we then poured it into a beaker with 2% milk to the 400ml mark using 40ml of rennet. We did this for rennet & milk, rennet & milk with yoghurt, and rennet & milk with buttermilk. My experiment dealt with, rennet & milk and rennet & milk with yoghurt. With the yoghurt, I just had a few tablespoons worth in the beaker.

After we had it in 37 degrees Celsius for a day, we then took out our results (see above) and drained each beaker to get the product. When that was done, we put the product back into the beaker and used Kosher salt. This looks like a Mexican cheese named cotija queso. (See right)

Then we have the cheese that looks like cream cheese for bagels. (See left)

All in all, making cheese isn't that hard but it's not easy either.

Lab Report Rubric

Registration Info for Next Term (Spring 2011)

Feed microorganisms oil so they can be full

Bioremediation

 Future plans for past, present and future disasters.

I was given an Oil Spill Bioremediation Kit to investigate that microorganisms can use oil in water as a food source. I thought it was interesting because the first thing that came to mind was "Why isn't this being applied to oil spills that have happened?" However, reading the manual+guide shared information about how the process of bioremediation has been implemented to oil spills, which saves the lives of sea animals, land animals, humans and marine life.Bioremediation is an interesting concept that I was introduced to since it is a way to clean up pollutants.

The question that I was given was, How can bacteria consume oil, and also how can they be beneficial when it came to oil spills?

My hypothesis was, Bacteria helps with oil spills by ingesting some factors or components of the oil, which as a result decreases the mass of oil. 

I learned some terminology that I defined which are the following:

• Bioremendiation refers to the use of living things to clean up environmental pollution. 
• Biodegradation is the breakage of oil over time that turns into simpler, nontoxic products by oil-degrading microbes. 
• Bioaugmentation is the process of supplementing (can be seen as "seeding") a population of naturally occuring, oil-degrading microbes with additional microorganisms. (This technique is used when the existing population of microbes in a contaminated region is not optimally suited to degrade the type of oil present. 
• Microbes are microscopic living organisms. 

 Excerpt from kit guide

 "Scientists recognize great potential in utilizing the oil-degrading properties of microbes to expedite the breakdown of harmful oil from spills."

A venue to the future...

As scientists continue to explore, especially with this process, it could lead to different things to clean up instead of oil. For example, everyday Americans put items in their trash, and that trash can goes to landfills. What if, if there aren't already, bacteria that can eat the trash to reduce pollution and garbage in our environments? Bioremediation is definitely something that I would like to continue to pursue and become more familiar with. 

12/6/11

Today I worked on getting my internship paperwork done and then i checked the PH on The Wine I made and then wrote it down in my notebook and the I labeled them to remember which is which.

11.29.11 - 12.01.11

Today we made..... cheese! We had to pasterize the milk (heat up) until 80 degree celsius then let it cool to 40 degrees. This wasn't the funnest process but we got it done. When it got to 40 degrees I had to add a spoon of yogurt and 40 mL of rennet solution. I then had to incubate the soon to be cheese at 37 degrees celsius. And thats it.