BIO250 Lab 2 micro pdf..

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Jan 9, 2024

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“ Lab 2 Culturing & Aseptic Technique BIO250L ” Student Name: Brandon Estelle Access Code (located on the lid of your lab kit): AC-GIF4SU Lab Report Format Expecta0ons U"lize college level grammar and professional forma4ng when comple"ng this worksheet. Submissions without proper forma4ng, all required photos or suﬃcient responses will be rejected. Pre-lab Ques>ons 1. This lab includes two experiments that each look at different aspects of culturing microorganisms onto growth media. What concepts does each experiment focus on? Why do you think these concepts are important to the field of microbiology? THE CONCEPTS EXPERIMENTS FOCUS ON ARE VIRUSES, BACTERIA, ALGAE, ETC: 2. In this lab, the experiments will allow or call for the use of four inoculaVon tools. List these tools and provide a scenario in which you would use each tool. ○ Pipets ○ Agar plate ○ InoculaVng loop ○ Culture tube 3. Describe a piece of equipment used in biotech labs to extend the growth pa\ern of microbes. Focus parVcularly on those used with E. coli used to make human insulin. Incubators because they help bacteria cultures grow at ideal temperatures. 4. You’re a physician trying to isolate bacterial colonies from the human gut in an a\empt to diagnose a gastrointesVnal infecVon. You streak your sample on a growth media containing glucose, amino acids, and salts containing both sulfur and phosphorus with a pH of 7 . You incubate the plates in aerobic condi>ons at 37 ˚C for three days, at which point you can see clear bacterial colonies forming on the plate. Would you feel confident in staVng that you had successfully cultured all the bacteria from your gut sample? Why or why not? No personally I would not feel comfortable trading the paVent because I would only be seeing the aerobes on the culture. Also, because the anaerobic bacteria will not grow in this experiment. EXPERIMENT 1: AGAR PLATE PREPARATION AND BACTERIAL INOCULATION Introduc>on Ques>ons

“ Lab 2 Culturing & Aseptic Technique BIO250L ” 1. Describe the objecVves of Experiment 1. State the goal of the experiment and the learning outcome that it is trying to convey to you as a microbiology student. The main objecVve is to see how much bacteria growth can be on regular household use items. 2. Proper asepVc technique is crucial to ensuring the growth of pure bacterial colonies. What will you do in this lab to demonstrate that you are using proper asepVc technique? Proper hand hygiene, no food or drinks near lab, using the provided sterile equipment. In a laboratory sebng, what are three ways you can properly sterilize culturing equipment? ○ Dry heat ○ Wet heat ○ UV radiaVon or chemical solvents 3. What method of sterilizaVon will you use in this experiment? Dry heat 4. What results do you expect to see with this experiment? For example, do you expect to see growth from all surfaces or just some? Are there parVcular strains you expect from the surfaces you swabbed? I expect to see growth from some heavily used areas but not all as to some are kept clean. I expect to see more growth on the shoe bo\om than anything.

“ Lab 2 Culturing & Aseptic Technique BIO250L ” Data and Observa>ons In the table below, report your observed growth corresponding to each plate number and its corresponding source. Then provide a photo of your growth plates. Your data should have a quanVtaVve aspect to it, such as a count of the number of colonies that grew. Your data must also correspond to the photo you include for credit. Table 1: Colony Growth Provide a clear, high resoluVon photo of your plates ader incubaVon with the lids removed. For credit on this lab, your photo must show the growth that is reported in your tabulated data and must also include your handwri\en name in the background. Plate Number Source Growth (Color, Amount, Shape, etc.) 1 Teeth 0/ no growth 2 Fridge handle + Li\le growth round yellow, white colonies 3 Shoe bo\om ++ moderate growth about 7 colonies yellow/white 4 Control 0/ no growth 5 Airborne 0/ no growth

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“ Lab 2 Culturing & Aseptic Technique BIO250L ” Sample 1 Sample 2 Sample 3 Control Airborne Contaminatio n

“ Lab 2 Culturing & Aseptic Technique BIO250L ”

“ Lab 2 Culturing & Aseptic Technique BIO250L ” Results and Discussion 1. Describe what morphological traits (i.e., size, shape, arrangement, color, margin, etc.) you observe from the growth on your plates. Be as specific as possible in the characterisVcs. The fridge handle sample grew in white, yellowish, circular shape, there was only one circular gross on the enVre plate. It was smaller than a dime. Usually the bacteria that grows on a fridge handle is micrococus, bacillus species, staphylococcus epidermidis 2. Based on the morphological qualiVes you observed on your plates, and using any resources available to you (i.e., a textbook or internet sources such as the CDC website), look up what bacteria you may have found on the surfaces you swabbed. I believe i grew staphylococcus epidermis on the fridge handle due to it being touched daily by humans and skin cells which carrier bacteria. 3. For the colonies hypothesized in QuesVon 4, are you surprised to find this type of microbe on this surface? No i am not surprised since this typed of bacteria growth is found on humans 4. Looking at your control, did you perform proper asepVc technique or were your plates contaminated? If contaminaVon did occur, list the possible sources and how you can prevent

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“ Lab 2 Culturing & Aseptic Technique BIO250L ” contaminaVon in the future. In my opinion, I believe I did perform proper asepVc technique on my plates. My control was not contaminated. There was no growth on the Agar plate, and I was able to do the control fast and cover the plate to prevent any bacteria. 5. Compare the colonies that grew on your swab plates to the Airborne ContaminaVon plate. Did any of your experiment plates grow colonies similar to your Airborne ContaminaVon plate? How would you describe the risk of airborne contaminaVon in your experiment? There was no growth on my airborne contaminaVon plate, so these cannot be compared “

“ Lab 2 Culturing & Aseptic Technique BIO250L ” EXPERIMENT 2: BACTERIAL TRANSFER TO STAB TUBES AND AGAR PLATES Introduc>on Ques>ons 1. Describe the objecVves of Experiment 2. State the goal of the experiment and the learning outcome that it is trying to convey to you as a microbiology student. Click here to enter text. 2. What results do you expect to see in Experiment 2? Click here to enter text.

“ Lab 2 Culturing & Aseptic Technique BIO250L ” Data and Observa>ons Record your observaVons in the tables below. Table 2: Ini>al Reserved Plate Colony Growth Observa>ons Table 3: Final Plate and Stab Tube Growth Observa>ons Plate Sample Appearance of the original colonies (morphology, etc.) 1 MulVple lines from inoculaVng loop 2 Yellow dots from inoculate Loop 3 White and yellow dots Sample Form Growth (Y/N) Same Appearance as Ini>al Plate (Y/N) Successful Transfer? (Y/N) 1 Plate Yes No Yes 1 Stab Tube Yes No Yes 2 Plate Yes No Yes 2 Stab Tube Yes No Yes 3 Plate Yes No Yes 3 Stab Tube Yes No Yes Control Plate Yes No Yes Control Stab Tube No No Yes

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“ Lab 2 Culturing & Aseptic Technique BIO250L ” Arrange your plates and tubes as shown below. Remove the lid from the plate and place the stab tube next to the matching plate. Then take a high resoluVon photo and insert it below with your handwri\en name in the background. For credit on this lab, the growth that you report in the above table must correspond to the photo below, and the photo must be of a high enough resoluVon to assess the reported data. Plate/Tube #1 Plate/Tube #2 Plate/Tube #3 Control

“ Lab 2 Culturing & Aseptic Technique BIO250L ”

“ Lab 2 Culturing & Aseptic Technique BIO250L ” Results and Discussion 1. Review your data and determine whether or not the growth on your transfer plates have the same colony morphology (i.e., size, shape, arrangement, color, margin, etc.) as your original sample. Discuss this below. Yes, they do have the same color. Just the size is a li\le bit bigger. 2. If there was any difference in colony morphology, what do you think could have caused this? More bacteria growth

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“ Lab 2 Culturing & Aseptic Technique BIO250L ” 3. Imagine you are performing an experiment that requires the transfer of cells to different plates which will grow bacteria over an extended period of Vme (days or weeks). Describe why an unsuccessful transfer or growth with a different appearance from the iniVal plates would be a problem in this scenario. 4. Do some research and describe two or three scenarios in which it would be preferable to use a stab tube vs. a growth plate. ( Hint : What do bacteria use to help them move? Can moVlity be used to help idenVfy many medically important pathogenic bacteria such as the Enterobacteriaceae family of bacteria?) The growth plate allows more oxygen to be led in growth rather than the stab tube

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