Bacteria [Communications]

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Trader
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Project Question: Role of AI-2 in Biofilm Formation of L. monocytogenes
Project Due Date: Janurary 1, 2010 (2 months!!)
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Re: Bacteria [Communications]

Post by Trader »

The first pair of judges noted that how I organized the tables (it was a bit last-minute prepared) was not the best way to display the richness of my data?

But the second pair of judges said that they really liked how I organized the data! o=. But either way, I think I should reorganize my data.

I'm hoping that I can do more trials, or even better, measure the oxygen levels that there are?... But I don't think I can do that all in one month :(.

Perhaps just a little more trials will be good. I really want to measure oxygen levels as well.

I just looked at the projects the previous ISEF finalists had...they are so fancy! Then I looked at the judging guidelines -- in what section would my relatively un-fancy project have marks taken off of?

xD either way I'm very excited to be part of the experience. Sounds like so much fun!!

Next year I'll go fancy ... in addition to understanding =P
Trader - scientist wannabe =)
donnahardy2
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Re: Bacteria [Communications]

Post by donnahardy2 »

Hi Trader,

I think you definitely need to redo the data, but I need a chance to look at it again before I can give you specific advice. It would be nice to repeat the standard curve at 30 and 37 degrees C, and add the OD at 660 nm (I can't remember-do you have a spectrophotometer?). But I definitely don't want you to work up to the deadline again. Since you are going primarily for the experience this year, it would be perfectly OK, just to redo the graphs that you have and make the board as presentable as possible. I would say you should leave at least 2 weeks for the board, so if you can't start an experiment next week, don't try. It's really hard to do this type of work without new sterile Petri dishes and pipettes.

The science buddies website has a forum for students who are entering the ISEF fair. I'm not sure if you should start a new post in that forum; I think it would be better to continue this project here, but I've sent a message to the moderator of that forum to find out. I would be good to get advice from someone who has participated in that fair.

Hopefully we'll have an answer tomorrow.

Donna Hardy
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Re: Bacteria [Communications]

Post by Trader »

The school does have a spectrophotometer :D.

In the one month that I have, I probably would want to do more trials to make sure that my conclusions are stronger. My current title is "Analyzing growth of facultative anaerobe bacillus subtilis under microaerophilic conditions" --

i just want to make sure: would it be expected of all facultative anaerobes to have lower growth rates (longer lag times, doubling times) under microaerophilic conditions?

I know in my hypothesis I expected the highest amount of viable organisms to be lower, but it turns out that wasn't true, given my platings.

I expected I wouldn't get anything at the regional fair so I already packed everything up and threw all my preparation materials away :(, but it won't take more than a few days to get prepared again.

is there an "easy way" to measure available oxygen levels? And for my significance, I told many judges that doubling times/growth patterns would be important in determing the creation of industrial enzymes, but I never really knew anything beyond that -- perhaps there are some special conditions (so and so oxygen levels with the addition of so and so) that I can add (unless it'll take too long) so that I can see the new growth?... Ahh I don't know :).

I guess the idea of me representing the region is just hitting me <_<. The judges said that the other finalist and I (both from the same school :D) had a chance at getting "a placing", and I guess I don't want to let them and the region down...I think the chances are probably not there ,but I'll want to do my best :).

Thank you for looking into it!
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deleted-71588
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Re: Bacteria [Communications]

Post by deleted-71588 »

Congratulations. You've come a very long way on your scientific quest for knowledge. In fact, so far that you are now just beginning to understand that with every discovery come more questions. DO NOT GET DISTRACTED at this critical point!

Donna gave you some excellent advice already and I'd like to say it again in slightly different words after stating the KISS principle: "Keep It Simple Stupid!". You had an excellent project that got you this far. Don't screw it up and start changing things and investigating additional things. Hopefully Donna will get back to you soon after she has had time to take a second look at your data with specific recommendations.

In any case, the biggest difference in the ISEF level of competition from the regionals is in presentation materials. You've done the "science", now it is time to show that you've done the science and that you have the skills to present it. Your audience will be a wide range from those that may or may not have an interest in your specific area of investigation to those who are extremely knowledgeble in your specific area. You need to display it so that people walking by your booth are drawn in to take a closer look. You are literally "selling" your project to the passer's by. In fact, you maybe selling yourself as a young scientist to potential organizations and people who might be interested in helping you along your path to scientific knowledge and education.

Research scientists not only have to do the science, they have to sell their projects, their scientific integrity, and their interests to organizations that fund research. This is an excellent opportunity to hone your presentation skills. Make good use of this opportunity to present yourself and your project. You would not have gotten this far if it wasn't worth presenting well!

Again, congratulations.
-Craig
donnahardy2
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Re: Bacteria [Communications]

Post by donnahardy2 »

Hi Trader,

Craig has given some excellent advice based on his experience with the ISEF level of competition. One of the problems with getting data on any experiment is that the results will point to 10 more experiments that need to be done, and you are definitely thinking about doing too much in a short time. At the ISEF fair, you need to present your data like one of the scientific journal articles that you read last fall, so the presentation and analysis are critical. There are problems with your data that are going to affect the judges' evaluation of your data, so it would be nice just to repeat the 30 and 37 degree growth curves to replicate the results. But think about it and decide if you can do this fairly easily. Otherwise, just plan to spend your time on redoing the presentation with the data you have. Your project is complete and there are lots of details to discuss.

Unfortunately, I won't be able to review the data thoroughly until this week-end, but I'll try to send some suggestions just as soon as possible.

Donna Hardy
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Re: Bacteria [Communications]

Post by Trader »

OK. I'll repeat the growth curves, and then focus on the presentation.

For the flaws in my data, pointing out what I know/what I did wrong -- that won't help as much in the ISEF right? Because I think that in terms of science and skill, I basically relied all on science to qualify ... and that's not going to help me as much in the Intel ISEF <_<.

I will use the spectrophotometer to see how my dilution plating (I'll do one more plating for 37, one more for 30) correlates with it.

A few things I was curious about --

is there any way to measure oxygen levels? (I'm not going to attempt the linear relationship, but I really want to make it less ambigious that my environment is microaerophilic [it was one of the questions the judges asked and I wasn't really too sure about the question])

For the skill emphasized in science fairs, does that translate to "fanciness"? (That's how I'm interpreting it ><...)

Finally, for my hypothesis (I speculated that it would have longer lag phase, longer doubling time ... ) -- was it a given that any facultative anaerobe would have that lower growth rate in a microaerophilic environment?

Sorry -- these are the last complications I'll be pursuing! And I'll research more about the why the results happened.
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deleted-71588
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Re: Bacteria [Communications]

Post by deleted-71588 »

For the flaws in my data, pointing out what I know/what I did wrong -- that won't help as much in the ISEF right? Because I think that in terms of science and skill, I basically relied all on science to qualify ... and that's not going to help me as much in the Intel ISEF <_<.
WRONG! Everything you have done so far is still VERY VERY important. Everything that has mattered in all of the preceding competitions still matters. If you loose any of what mattered, you lost something that made your project a great Science Fair project. It is just that at the ISEF level, you need to add polished presentation materials. You still have to prove that you did a great Science Fair project. The subtle difference is that you also have to prove that you have learned how to present it as well.
is there any way to measure oxygen levels? (I'm not going to attempt the linear relationship, but I really want to make it less ambigious that my environment is microaerophilic [it was one of the questions the judges asked and I wasn't really too sure about the question])
When I've have judged Science Fair projects in the past, I always asked at least one question I didn't expect the participant to be able to answer. I've done the same thing in every job interview I've ever conducted. This allows me to judge character and integrity and the ability of the participant to think under pressure. I remember back to an individual I competed against who everybody thought was brilliant and would go far. Many years later, that individual was caught faking data on a research project which was a carear ender and a huge disgrace for the employer and it called into question large amounts of reasearch done by the organization. If that individual had learned the lesson that it is perfectly acceptable to answer with a simple "I don't know", things might have turned out a lot better. Often knowing what you don't know is more important than knowing what you do know.

"microaerophilic" is a property of a bacteria or organism which by definition means that organism requires oxygen to multiply; however, its growth is NOT impeded by an atmosphere that contains less O2 partial pressure than the normal earth atmosphere at STP.

There are ways to measure oxygen levels but they are expensive in terms of the equipment needed and difficult in terms of controlling equipment contamination so they typically aren't used for your kinds of experiments. The two common methods for insuring an environment to prove an organism is microaerophilic are 1) introduce a competing gas such as CO2, NO2, or helium and 2) reduce the atmospheric pressure with a partial vaccum but these methods require a sealed environmental chamber. In order to classify an organism, you have to attempt to grow it in aerobic, microaerobic, and anerobic environments to compare the growth and growth rates.
-Craig
donnahardy2
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Re: Bacteria [Communications]

Post by donnahardy2 »

Hi Trader,

Craig has made some excellent comments about your data presentation. The data you obtained is real data (empirical results), so you can't change the data. You want to present it in a positive way and explain it logically. I should not have said your data had problems; I should have said it would be challenging to present and explain the results.

Oxygen analysis is very difficult and requires instrumentation, and the values would not add that much to your results. The addition of an OD reading at 600 nm might possibly verify the results, or help explain why the generation time was so short on your first experiment and would not be that difficult to do.

Here is the site that I tried to show you before, that you could not open. I printed it and scanned it so hopefully you can open it now. It would be great if you could present your data in a graph form as well as the table format. And the graphs shown here show the approximate relationship between viable plate count and OD at 600 nm.

Maybe someone has published results of growth in Bacillus species under microaerophilic conditions. If you could find a literature reference, you could compare your results to the published data for an interesting analysis. I haven't looked yet for this type of reference, but I'll see what I can find.
bacterial growth curve.pdf
how to graph a bacterial growth curve
(208.89 KiB) Downloaded 428 times
Donna Hardy
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Re: Bacteria [Communications]

Post by Trader »

Thank you for your replies, Craig and Donna :D

Ahh, I see now. Because the presentation is the biggest difference between the two (as well as the level of the projects), I'll continue what I'm doing before, only this time by "better presentation skills", does that mean that I should also demonstrate a greater depth of understanding of the results, and actually include things that I should have included before like error analysis, a better presentation in terms of the board, bringing all my data into questioning, like that?

Another small question would be -- how much does how "fancy" a project look like factor into the judges' decision? such as, if there were two equal presenters with just as equal of an understanding and presentation (if I ever get there <_<), how much of an advantage would the "fancier", more "advanced" science project have? -- I realize that this is probably not the direction you want me to be thinking, but it's something I can't really get off my mind.

As for the microaerophilic conditions, nutrient broth w/o aeration is definitely OK right? Sorry, I did ask this before, but I'm worried about the actual definition of microaerophilic -- since aerobic is 20% oxygen, and microaerophilic is around 2-10%, I wonder if the oxygen level would be 11-19% and/or I'm really only measuring the growth curves of the bacteria that are closest to the air in terms of the nutrient broth w/o aeration, and not those at the really bottom ...

THANK YOU for the bacteria growth curve pdf!! I was looking at it when I was finishing up the graphs the night before the fair <_< and I was attempting to use semi-log paper on graphical analysis, but gave up in the end and prayed that I wouldn't be asked about my graphs later.

Thanks for all your advice! >< I definitely couldn't have gotten far without all this help.

From my understanding of what I'll see in the ISEF
- Spectrophotometer to analyze the doubling time.
- More observations on how long it'll take to get turbid (with a constant volume to reduce that error, though mention this before)
- More trials regarding highest amount of viable organisms.
- Stop at two weeks before the fair!
- Then work on all the presentation (thanks once again for emphasizing this!)

This is from my understanding of the difference now :D.

Now I remember that I should have asked the judges at the regional fair how I could have improved!! Now I could only ask by email, which is not good <_<. Perhaps I can talk to them a month later on the plane, but...

THIS IS SO EXCITING ;D
Trader - scientist wannabe =)
deleted-2131
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Re: Bacteria [Communications]

Post by deleted-2131 »

Hi Trader,

CONGRATULATIONS!!! I am very excited for you and wish you best of luck. As you are already sensing, preparing for ISEF is very important. Your goal is to be prepared with both professional-quality research and a professional level display and presentation. A significant portion of judging is done without you there, so your board needs to be clear, complete, cohesive, and memorable. Your abstract is also critical because many organizations will decide whether or not to judge you based on your abstract. Your judges will be evaluating many, many projects, so your research and your presentation need to stand out in their minds.
Because the presentation is the biggest difference between the two (as well as the level of the projects), I'll continue what I'm doing before, only this time by "better presentation skills", does that mean that I should also demonstrate a greater depth of understanding of the results, and actually include things that I should have included before like error analysis, a better presentation in terms of the board, bringing all my data into questioning, like that?
Yes.
Another small question would be -- how much does how "fancy" a project look like factor into the judges' decision? such as, if there were two equal presenters with just as equal of an understanding and presentation (if I ever get there <_<), how much of an advantage would the "fancier", more "advanced" science project have? -- I realize that this is probably not the direction you want me to be thinking, but it's something I can't really get off my mind.
I'm not sure what you mean by fancy or advanced. Your display needs to be well organized from a content point of view and well planned from a graphic design point of view. It needs to be able to stand alone and tell the story of your project without you there to explain it. You should have the most important parts of your project emphasized - the buddy board idea works well. There is a balance between too much text and not enough information, and this can be hard to find. The importance of your project and its applications should definitely be highlighted. Awards will go to the students with three characteristics (1) students with solid science (sound methodology, lots of data, substantial data analysis, logical conclusions and meaningful applications), (2) students with a deep understanding of their subject matter, including both the very germane and the somewhat broader field, and (3) students who can sell their project - their display and presentations are top notch and memorable. You should be able to speak confidently without lots of umms... and ahhhs... and be able to answer questions intelligently. It's helpful to pause for a moment after a judge has asked you a question to gather your thoughts. Repeat the question to make sure you understand it, and then answer it. Taking these few moments will allow to ensure you understand the question and will help you construct a cogent answer. Like Craig said, it is important you know your limits - if someone asks you a question you don't know then say "I don't know".

As has already been said, Science Buddies has a wealth of resources to help you prepare and I strongly suggest using them! Donna, Craig, and I are here to help as well and will continue to guide you as best we can. Keep your questions coming and we'll keep answering them. In the coming month, try to focus more on what you can do and what you have done and less on other competitors and hypothetical scenarios. This will keep you focused, driven, and much less stressed. You have a great project! Keep working hard and you will succeed.
All the best,
Terik
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Re: Bacteria [Communications]

Post by Trader »

Thank you for clarifying that!!

Wow...This means I should definitely do some background research :D:D

The fanciness was referring to the complicated sounding titles that the winning projects had, but I now know to 1) not focus on hypotheticals as well as 2) know that knowing of background info is more important.

I tend to focus a lot on how others are doing ><. A lot of the psychology impacts me, unfortunately, but thank you all for the confidence boost :D.
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Re: Bacteria [Communications]

Post by deleted-2131 »

Trader,

I think you probably know more background information than you think you do. All that reading of scientific papers - that's background research! I would focus on your remaining experiments, data analysis, and display/presentation before focusing on more background information. Stay focused on what's most important - your list a few posts back is a good one in my opinion (although Donna is a better person for the actual science goals than I am - I'm a planetary scientist, not a biologist.)

With regards to titles: they only sound fancy because you don't understand them. For example, if I said that the title of my project was "Performance Analysis of the Lunar Time-of-Flight Mass Spectrometer: a Novel Approach for the Complete Characterization of Transient Gas-Releasing Events", I'm guessing you would call that fancy. To someone who knows the subject, it makes perfect sense. Your current title, which I believe is "Analyzing growth of facultative anaerobe bacillus subtilis under microaerophilic conditions" seems quite fancy to me because I don't understand microbiology at the level that you do.
All the best,
Terik
donnahardy2
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Re: Bacteria [Communications]

Post by donnahardy2 »

Hi Trader,

Thanks to Craig and Terik! Your comments are very helpful.

Craig and Terik have given some very excellent advice for presenting your project and for preparing for ISEF. I'm sure you will be able to incorporate their suggestions as you focus on finalizing your project. You should just concentrate on making your project the best that it can be.

I have just printed out your data again, and I will take a look at it now that more time is available. I have gone back through the posts just prior to the first science fair, and I can't find anything except the data. Would it be possible for you to post the rest of your presentation? Or, if you have done this, just tell me which date you posted it and I'll find it. As Craig and Terik have mentioned, an understanding of the science and an analysis of the data are critical at ISEF, so it would be helpful to see your whole presentation.

I know you are planning to repeat one more growth curve, so please ask if there is anyway you could have time and access to the lab so you could take your samples and plate them at the same time. From your data, I think that the bacteria didn't stop multiplying when you took your samples; the organisms were in log phase and apparently had lots of momentum and kept dividing until you were able to plate them. I am not suggesting that you do anything that would interfere with your other classes, but perhaps an adult could stay with you in the lab a little later, or on a Saturday to do the growth curves. If this is not possible, then redo the experiment exactly like you did it before and we'll see if the results are reproducible.

If you have not used a spectrophotometer before, ask if you can look at the instruction manual. Find out if cuvettes are available to use. For 600 nm, you can use disposable plastic cuvettes, and this will save time, compared to quartz cuvettes, which have to be cleaned in between each reading. If you include spectrophotometer results, you will need to understand how a spectrophotometer works, and why 600 nm is used to estimate numbers of bacteria. Please let me know what brand and model of spectrophotometer you have available to use.

I have just a couple of comments on the project board. It is helpful to print out the pages and lay them out on your board and stand back to look at the proportion between the words and spaces. This is very important in making a board that's easy to read, and you can't see this until you try it. A type-size that's too big or too small for the space will not be as easy to read. So you will have to find the balance that Terik mentioned between "too much text and not enough information," that will be easy to read. Try to make the presentation balanced and symmetrical and do make sure it is easily readable. Anything visual that you can add to help the judges understand what happened will be helpful. For example, a flow diagram of your experiment. Or, a diagram showing why your culture was growing under microaerophilic condition. Or, perhaps a drawing of a Gram-stain that you did or a photograph of the equipment you used. Or a drawing of a little rod with cilia to show any non-microbiologist judges what your subject looks like. Your visual information will help make the project memorable to judges who are looking at a lot of projects in a short amount of time and help them "see" what happened. Adding a little color to the board is helpful, for example, as a border. Use bright, clear colors that are monochromatic or complementary and your project will be very memorable. I don't know if ISEF is judged on a point count, but for other fairs, there are a few points that are awarded for the quality of the presentation. I imagine that in a fair like this, there is very little difference in the point count of winners vs non-winners, so an extra half-point awarded for the presentation could make a difference.

Donna Hardy
donnahardy2
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Re: Bacteria [Communications]

Post by donnahardy2 »

Hi Trader,

One more thought. If you haven't heard back from some of the internship programs you applied for, you could consider sending an update to each program to let them know that you are an ISEF finalist. It might help.

Donna
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Re: Bacteria [Communications]

Post by Trader »

I think my board is OK (well, the left panel at least) -- I think I can get the photos uploaded by next post, because I know that the cameraman took a picture of it.

This is exactly what went on the board (some of it is last minutely prepared, please understand :D)

[The extension doc has been deactivated and can no longer be displayed.]

>> OK, looks like only 1 attachment is possible so I'll be making multiple posts -- they're also in big font because they were the size I used to put on the poster.

My science teacher suggested that I immediately go to the board, but I convinced him to give me a week. For the spectrophotometer, I would spend a Saturday where the morning I would inoculate the bacteria, and then every hour I would take one sample
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Re: Bacteria [Communications]

Post by Trader »

Data:

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As for the data, I know that the uncertainty with dilution plating is much greater than when using the spectrophotometer. If I were to get results from the spectrophotometer, would I want to shift my conclusion and analysis primarily on that obtained from the spectrophotometer because it is so much more accurate, or would it be depending on whether there is a close correlation between the values obtained for each?

IF I inoculate nutrient broth in the morning and get 80 minute intervals until late in the night, would that be enough? I'm a bit worried because it seems like the stationary phase won't come until 12 hours or so after incubation, which means that I'll have to stay very late...

Oh no... but I can't do that during the day because I'll have to leave them out... <_<.

OK, I'll have to use the Saturdays then. I wish I had separate incubators!

One more thing! For EVERYTHING, there is an uncertainty of half the smallest increment right? So even if I was using machines with electronic readings (is this what a spectrophotometer does?), there is always uncertainty right?

From what I understand, I would want to plate the samples at 30 and 37 at least...one of the first things on my to do list is to find out what conditions bacillus subtilis are used in when creating industrial enzymes. If there are certain temperatures they are usually created in, I would want to measure those right? [I'm worried about the significance of my project...sorry -- maybe I'm worrying about the wrong thing]

(For its use in probiotics, the stomach would be a microaerophilic environment right? And that would be 37 C, so that's cool)

I just want to clarify -- oxygen levels cannot or should not be measured because I won't have the tools (I don't think I do) or the time right? I was thinking that there might be some coefficient that has a linear regression with the amount of oxygen levels. If I can somehow measure the oxygen level available in my "microaerophilic condition", and then somehow do "normal" growth curves by having an incubator and stirrer working the entire time at a high heat that results in 30 or 37 given the unclosed incubator door...and then find if there's a linear relationship.

But I can already tell by the description that I'll never be able to get that done in time. Awww...

For now it looks like: growth curves (two Saturdays, or maybe one entire weekend) for 30 and 37 C, and then STOP.
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Re: Bacteria [Communications]

Post by Trader »

Graphs:

[The extension doc has been deactivated and can no longer be displayed.]

These were poorly done... I definitely know this.

I should have the axis constant, and I should have definitely somehow graphed the (hopefully) constant growth rate (especially for 30) with semilog y axis.

I should have added error bars (?) or at least indicated my uncertainty somewhere.

One of my graphs (for the aerobic conditions) was also completely based on the assumption (unsupported by any literature because it was last minute :() that the doubling time would be 20 minutes ... <_<. The problem is, when I type in Google "bacillus subtilis growth aerobic", I can't get any ASM journal type (or related) studies on its growth. I've only found a few studies mentioned a doubling time:

[link]http://www.pubmedcentral.nih.gov/articl ... tid=214739[/link]
[link]http://jb.asm.org/cgi/reprint/179/10/3371.pdf[/link]

I'm afraid I really have no support about the doubling time of bacillus subtilis. I contacted the authors for those who investigated the growth of bacillus subtilis under anaerobic conditions if that might help, but just how different the conditions my experiment was setup in (such as the special type of nutrient broth) worries me a bit.

I'll continue hunting for it -- a really cool modeling study of e. coli growth had models of its growth to compare with, and I hope that perhaps someone modelled b. subtilis growth before? It's really cool.

I really want to do the below for b. subtilis ... at least part of the below ^^:

[link]http://rms1.agsearch.agropedia.affrc.go ... 7-3029.pdf[/link]
[link]http://aem.asm.org/cgi/reprint/71/12/7920.pdf[/link]

But I can't seem to find any of the above for any other species but e. coli :(. I'll keep searching...
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Re: Bacteria [Communications]

Post by Trader »

Discussion:

[The extension doc has been deactivated and can no longer be displayed.]

I know that with the addition of growth curves, I would definitely be able to discuss more.

My teacher appears to be reluctant for me to continue my project -- I am pretty sure I will be able to conduct growth curves using the spectrophotometer, but (sorry if I overlooked anything) would it be "worth it" for me to continue to plate samples (even when I can access the more accurate alternative of the spectrophotometer?) -- if I can, I know I can reinforce the data that the dilution plating appears to give.

My abstract paperwork is due in approx 5 days and my teacher wants it in soon -- I know that the weekend isn't until another 5 days, so I wouldn't be able to add what I learned from the spectrophotometer in the abstract. I'll ask about the details regarding updating the abstract (though I don't think they'll like that), but in general, would it be OK if the abstracts have "vague" conclusions such as "the lag times were found to be longer, the doubling times were found to be longer, and the highest amount of viable organisms at the stationary phase were found to be approximately the same as those reported in literature" -- would that be OK?

Because I hope that the spectrophotometer will emphasize what my dilution plating results appear to indicate. (my 37 degrees Celsius results are not so great, and my 30 degrees Celsius results appear to be a bit weird too ...)

In response to a few things Craig said:
"microaerophilic" is a property of a bacteria or organism which by definition means that organism requires oxygen to multiply; however, its growth is NOT impeded by an atmosphere that contains less O2 partial pressure than the normal earth atmosphere at STP.
OH. Does this mean that a facultative anaerobe by definition would have growth impeded by an atmosphere that has less 02 in the atmosphere than at normal levels? [I think this is meant by STP?] I think definition of b. subtilis as something that's not a strict aerobe was established in 1998, as for it being a facultative anaerobe I thought I saw another study showing that it is a facultative anaerobe...
There are ways to measure oxygen levels but they are expensive in terms of the equipment needed and difficult in terms of controlling equipment contamination so they typically aren't used for your kinds of experiments. The two common methods for insuring an environment to prove an organism is microaerophilic are 1) introduce a competing gas such as CO2, NO2, or helium and 2) reduce the atmospheric pressure with a partial vaccum but these methods require a sealed environmental chamber. In order to classify an organism, you have to attempt to grow it in aerobic, microaerobic, and anerobic environments to compare the growth and growth rates.
OH, so an environment between aerobic and anaerobic environment would not be microaerophilic, but would be microaerobic instead (or is there a difference between the two?) -- where microaerophilic only describes the properties of the organism's growth under varying oxygen levels?

Sorry if I overlooked any of this -- so then, my condition (nutrient broth plugged w/ cotton, no stir [is no stir equal to no aeration?]) is microaerobic?

Ohh I really want to be able to measure oxygen levels...but that is for sometime else. Bacteria growth is cool!

Thanks!
Last edited by Trader on Mon Mar 30, 2009 7:07 am, edited 1 time in total.
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Re: Bacteria [Communications]

Post by Trader »

Conclusion:

[The extension doc has been deactivated and can no longer be displayed.]

My favorite part is explaining why!!! :D

The metabolizing of nitrate and nitrite is really cool. I'll definitely look into it after I analyzed the data (starting with error! I was really worried that I didn't mention anything about error, only sources of uncertainty, in my presentation) ...

To do list right now is to get growth curves in 30 and 37 C under spectrophotometer. :D Of course, the paperwork.

Thank you so much for everything!!! I am so excited about this. It's probably the coolest experience I'll have in my sophomore year, and definitely one of the most unforgettable ones in my high school life -- I can already tell!!!!!!!

XD

Abstract --

I've never written one of these before, and I've tried to get as much as I can from the abstract-teaching guides. I hope I did an OK job:
The experiment analyzed the growth of facultative anaerobe bacillus subtilis under microaerophilic conditions. Until recently, bacillus subtilis was characterized as a strict aerobe; studies concluding the bacterium’s metabolism of nitrate under anaerobic conditions found that this was not the case. The importance to analyze microaerophilic growth of bacillus subtilis is then apparent to test the classification of b. subtilis as a facultative anaerobe; because of the definition of this classification, it was hypothesized that the lag phase and doubling times would be between reported lag times and doubling times under aerobic and anaerobic conditions with similar experimental setup, and that the highest amount of viable organisms reached in the stationary phase under microaerophilic conditions would be lower than that under aerobic conditions. Quantitative observations to a turbidity standard were measured to record the lag time. Dilution plating and optical density measurements by the spectrophotometer calculated the doubling time and samples of the nutrient broth after 24, 36 and 48 hours of incubation were plated to record the highest amount of viable organisms at the end of the stationary phase. Recorded lag times were longer than that reported under similar experimental setups with aerobic conditions; calculated doubling times and generation times indicate a growth rate between aerobic and anaerobic conditions, supporting the hypothesis. However, the highest amount of viable organisms found at tested intervals of the stationary phase was found to be similar to that reported in related literature, refuting the hypothesis.
241 words?

I know my wording is terrible, I will definitely look into that later -- I wouldn't be penalized for vagueness right? It worries me that the judges might not even interview me if my abstract isn't good enough ... that I'll have to adjust to (I can definitely express verbally better than ... through writing) ... Kind of.

:D

Thank you once again!!
Last edited by Trader on Mon Mar 30, 2009 8:50 am, edited 1 time in total.
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Re: Bacteria [Communications]

Post by deleted-2131 »

Hi Trader,

Wow! That's a lot of posts and a lot of information for us to digest, so I apologize if I miss one of your questions in my response - just let me know and I'll answer it.

First - TURN IN YOUR PAPERWORK ASAP! Completing your abstract, the online finalist questionnaire, and mailing in your completed forms should be your most important priority right now. ISEF is incredibly strict about paperwork and deadlines. If you don't get things done on time, you will be disqualified - no ifs, ands, or buts about it. You've worked incredibly hard to get this far, so please, please get your paperwork done. You don't want to wait to the last minute to do it, either. I strongly suggest that you write your abstract and post it here for feedback before submitting it. Many judges - especially for special awards - will read your abstract and use it to decide whether to look at your project. If your abstract isn't superior (in content, grammar, style, spelling, etc.) then some of these judges might decide not to even look at your display, let alone interview you. It's OK if your conclusions are, to use your phrase, "somewhat vague". I think that the sample sentence you gave in your post would be just fine. Deadlines for abstracts are always far ahead of the competition or conference, so we can't always be as specific as we might want to be. There are also sections of the online finalist questionnaire that require writing and you should make sure that these sections are quality as well.

Second - I think that the time line Donna suggested for your project during this next 5 weeks is a reasonable one. Getting more data is important to helping you be successful; you also need to have ample time to analyze that data and put together your board. Stopping two weeks before ISEF to do the latter is reasonable. I can understand why your teacher would like you to stop experimenting and focus solely on the board and presentation, but I would recommend Donna's suggested timeline instead.

Third - Donna's suggestion about printing your board out on paper and laying it out to look at it is an incredibly important one. When I did my displays for ISEF, I would usually do a dry run about 4-5 times before I was pleased with it and then finally printed it for real. Her comments on using flowcharts, diagrams, and pictures are also excellent. I've attached an example of one of the panels from one of my old ISEF boards that shows one approach to integrating text, images, and flowcharts in the methodology section, which can hopefully give you some ideas to spark your imagination.

Keep up the hard work and keep the questions coming!

Terik
All the best,
Terik
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Re: Bacteria [Communications]

Post by donnahardy2 »

Hi Trader,

Thanks for everything. It's great to see the whole project put together. Your writing is excellent. I have printed everything out, and will read it tonight and post comments tomorrow. I have just a couple of suggestions on your proposed lab project. It's very ambitious and if you do 30 degrees C on April 4, and 37 on April 11, this would leave you with final data to analyze in less than a month before May 10. There is a definite advantage of stopping now and concentrating on the presentation, but redoing the experiment would offer the advantage of having duplicate results, with two experiments at each temperature You do have plenty to discuss with the results you have now.

Two comments on the abstract. Your abstract should refer to results from your first experiment, and you should not include, for example, "highest number," and optical density measurements in the abstract because these results don't exist yet. You can definitely add them to your presentation if you obtain more results, but you have to submit your abstract before the second experiment, and you don't know what will happen or if the second experiment will happen. The other comment is that Bacillus subtilis is a genus/species name so should be spelled with a capital B and printed in italics. You can make your abstract a little vague withand intriguing to leave room to include the second experiment results and inspire the judges to want to come and look at your project. How many words can be included in the abstract?

Microaerobic is a new term for me. I looked it up, and it seems to be used to describe organisms that preferentially grow in low oxygen environments, which is not Bacillus subtilis, which prefers to grow aerobically. Go ahead and use the term microaerophilic, and we'll make sure you have enough information to defend your use of terms in case you meet a judge who is an expert at microbiological semantics.

I'll post more comments tomorrow. Keep up the hard work.


Donna Hardy
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Re: Bacteria [Communications]

Post by deleted-71588 »

Trader,
Sorry if I confused you...

A "facultative anaerobe" is by definition a microorganism that grows equally well under aerobic and anaerobic conditions which means it will grow equally well with an excess of oxygen to an environment where there is no free oxygen.

STP (Standard temperature and pressure) based on NIST (National Institute of Standards and Technology) is 20 degrees C (68 F) and 101.325 kPa (14.696 psi). The only reason I introduced this term was that you can reduce the amount of oxygen available below that of a nominal earth atmosphere by creating a partial vacuum.

As Donna points out, your "Bacillus subtilis" prefers to grow aerobically (which means it prefers to grow in the presence of oxygen) so at some point between a normal earth atmosphere oxygen content decreasing down through a microaeophilic environment to one devoid of free oxygen, the growth rates will decrease significantly. A "microaeophilic" environment is one of those terms that is loosely defined and covers a broad range of oxygen levels.

Hope this clears up any confusion I added.
-Craig
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Re: Bacteria [Communications]

Post by Trader »

Thanks!

There is sure a lot of paperwork with ISEF...

Just to double check, "ISEF Questionnaire and the abstract" are the ONLY things that are required to be sent within 12 days of the fair, and is this only the electronic copy -- meaning we don't need to send the hard copy of forms within those 12 days?? I've asked the fair director and haven't had a reply yet -- it would be best to make sure, because the checklist tells us to have a hard copy (one to send to the society apparently), and another to send electronically.

Attached is final copy of the abstract -- some things edited :)
Until recently, Bacillus subtilis was characterized as a strict aerobe; studies concluding the bacterium’s ability to metabolize nitrate under anaerobic conditions found that this was not the case. This experiment analyzed the growth of Bacillus subtilis under microaerophilic conditions established by broth without aeration to test the establishment of the bacterium as a facultative anaerobe. Based on the definition of a facultative anaerobe, it was hypothesized that lag phase and doubling times under microaerophilic conditions would be between the lag times and doubling times reported under aerobic and anaerobic conditions, and that the highest amount of viable organisms reached in the stationary phase would be lower than that under aerobic conditions. Similar sized individual colonies from a streak plate were inoculated into nutrient broth and the time it took for the broth to reach turbidity as established by a turbidity standard were recorded. The doubling time was calculated through dilution plating and colony counting. Samples of nutrient broth after 24, 36 and 48 hours were plated to record the highest amount of viable organisms at the end of the stationary phase; two temperatures of 30 and 37 degrees Celsius were tested. Recorded lag times and doubling times were longer than times reported under similar experimental setups. The stationary phase population reached levels similar to experiments carried out under aerobic conditions
With abstracts -- is the idea to be as concise as possible, or to be concise as possible under the given word limit? There are definitely things I could add...
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Re: Bacteria [Communications]

Post by deleted-71447 »

Hi Trader,
I like the abstract. Is this actually "final" or are you still looking for suggestions?
Chris
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Re: Bacteria [Communications]

Post by donnahardy2 »

Hi Trader,

I'm glad Terik alerted you to the importance of the ISEF deadlines. Did you receive a copy of all of the rules for this fair, including size of the board, etc.? You should review all show rules and make sure you follow all of the requirements.

I like the new version of your abstract. It is straightforward and clear. I prefer the end of your first abstract, however, because it summarizes your conclusion about the doubling time under microaerophilic conditions. So if you have not submitted this, try to make the last sentence kind of a summary or conclusion of the whole project. If you have already sent this, it's very good and perfectly acceptable. If I were a judge, I would definitely want to come and see your detailed results.

I will post other ideas as I have time, one at a time. I know you are busy getting ready to set up your experiments and submitting the paperwork.

Here is information on McFarland standards, which are used as turbitity standards to estimate concentrations of bacteria. You do not have to make these standards, so this is for your information. If it turns out you can't use the spectrophotometer at 600 nm, however, you could use these to estimate numbers of bacteria, if barium and sulfuric acid are available. And if a judge happens to ask you about this, you'll understand the question.

http://en.wikipedia.org/wiki/McFarland_standard

Terik's procedure section that he posted was excellent. No wonder he won. Did you notice that you can look at the page and "see" how the protocol was done? And the very conservative border of color enhanced the presentation, without being distracting. Can you think of a way to present your method that is more visual? Just think about this for a few days; I'm sure you'll think of something.

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Re: Bacteria [Communications]

Post by deleted-2131 »

Hi Trader,

EVERYTHING NEEDS TO BE TO SSP WITHIN 12 DAYS OF YOUR FAIR. All the online things need to be submitted and all your paper forms sent in.

I'm sorry I didn't get to check this earlier to answer your question; I just finished up presenting at a conference, chairing the judging at a regional ISEF-affiliated fair, and finals are just around the corner. Whew! I'll try to check everyday, but there's no guarantees.

Donna's point about reading through everything to make sure you are in compliance is very important. I know that in the packet you received when you qualified for ISEF there is a lot of material and I know that you're extremely busy, but PLEASE read through all of it. In addition, I would suggest looking at the "Creating and Effective Project Display" PowerPoint, which can be found here: http://societyforscience.org/isef/index.asp (scroll partway down the page; it's under the STUDENTS & TEACHERS heading). You might also want to look at the last two pages of the official ISEF student handbook which can be found here: http://societyforscience.org/isef/document/hbk2009.pdf. Pages 8-10 of the official ISEF rules, which can be found here: http://societyforscience.org/isef/docum ... es2009.pdf.

Keep up the hard work, and keep the questions coming!
All the best,
Terik
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Re: Bacteria [Communications]

Post by Trader »

Thank you Terik for all the information! It clarifies a lot.

I was just out the past few days too! Went to an Asia-Pacific Activities Conference for a Quiz Team competition :)

Right now the Finalist login page has a complete set of ticks (complete, green)...and I followed the checklist and the Abstract, Finalist Questionnaire and ISEF Form copies are all in :D. I hope that's everything ^^

For the questionnaire, I kind of regret not putting in more, because from what I understand the judges will be using this information to base their judging on? The fair director gave me a contact email and I asked if I could add additional information.

There was a section asking us if the project involved anything in the following areas:

Resource Conservation
Sustainable Development
Pollution Prevention
Energy Conservation
Sustainable Development Explanation

I'm not sure about the first 3, but would the use of bacillus subtilis in industrial enzymes forming and fermenting be "energy conservation"? If they can still accept edits, I'll email them and hopefully they will add onto it?

As for the use of b. subtilis in the creation of industrial enzymes -- I can see what their use is in places such as;

http://www.rakuto-kasei.net/enzyme/natto.index.html
http://www.scialert.net/qredirect.php?d ... linkid=pdf
http://books.google.com/books?id=4VpZfx ... t&resnum=5
http://books.google.com/books?id=4VpZfx ... 5#PPA77,M1

There are many things I think b. subtilis does -- but I'm not sure if they are microaerophilic environments. It seems like many of the environments are actually anaerobic (according to my teacher), and I'm not sure I can find what environments the uses above are in -- is the process of using bacteria to create industrial enzymes usually anaerobic, or does it vary? Sorry I don't know much about the actual environments they're in, only what happens.
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Re: Bacteria [Communications]

Post by donnahardy2 »

Hi Trader,

Congratulations on getting the application turned in! It sounds like it was a major project to get the paperwork done.

I will think on it, but your project does not appear to be directly related to any of the conservation/energy conservation topics. It's more of a basic research project. Terik and Craig could probably explain the reason for the list of questions, but I suspect that it may be related to special awards. I'm sure the American Society for Microbiology will be judging for special awards, and your project will certainly be considered in this category.

I'll have time to post more tomorrow.

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Re: Bacteria [Communications]

Post by deleted-2131 »

Hi Trader,

Yes, the Finalist Questionnaire, the abstract, and forms are all the things you needed to do.

The information in the Finalist Questionnaire is used for a variety of purposes: collecting demographic information for statistical purposes, helping Special Awards judges determine if your project qualifies to be judged for their awards, and for the preparation of press releases should you win an award. If you can still update your information, then you are welcome to. However, if you have already submitted the online questionnaire then I'm not sure there is much you can do. You are welcome to email the contact person your fair director suggested, but I wouldn't to be able to add material to your responses if you have already submitted the questionnaire.

I would concur with Donna that your project does not fall into any of the environmental-related divisions you listed. But don't worry about it--you will still be considered for all the awards your project is eligible for.
All the best,
Terik
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Re: Bacteria [Communications]

Post by donnahardy2 »

Hi Trader,

Yes, I agree with Terik. You should not worry if your project does not fall into an environmental category; it's a perfectly good project in its own category. It's just not an environmental project. However, when you go to ISEF, do pay attention to the awards that are presented, and if you find that there are more scholarship awards in a specific category, then you could consider doing a project next year that would give you a better chance of winning a scholarship. For example, in our local science fair, there are always lots of physical science projects and hardly any math/computer projects, so I always recommend to students looking for a project to do a math/computer project because there's one blue ribbon given in each category and it's better to be one of two, rather than one of 67, or so. So, this is a possible strategy for winning. However, you should always do a project that you are really interested in, rather than one because you think it is a better way to win. Your project won't be successful unless it's intrinsically interesting to you. I hope this makes sense.

One comment on your write up. Your references should be cited in a bibliography section at the end, and indicated with numbers in sequential order with a superscript number. I don't know if ISEF has a standard bibliography format that is suggested. If not, then pick a standard format and use it with all of your references. For example, this is the format that I usually use: "Hunter, et. al. J. Chromatog. A, 897, 65-80, 2000." The J. Chromatog, of course should be in italics. It is acceptable, and I think preferable if you have room to include the title of the reference. Having a perfectly done bibliography will enhance your project.

Donna Hardy
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