Pre-Lab Lab 1: Microbiology Lab Safety (100 points) Student Name: Student ID: Course ID: BIOL202 -Each question on the lab worksheet must be answered completely, thoroughly, in complete sentences and correctly in order to be considered for full credit -If the question asks you to do research or find a source, a reputable, credible and/or scholarly source citation must be included in order to be considered for full credit -If a math formula is required to arrive to an answer, work must be shown otherwise, no credit will be awarded Questions 1. What constitutes personal protective equipment? When should personal protective equipment be worn? Why is personal protective equipment important? (5 points) Personal protective equipment (PPE) consists of safety glasses, gloves, apron or lab coat and an underpad. Depending on the (BSL) microbes and chemicals being used, additional PPE may be required. However, the items referenced above are required for the lab experiments in this course. Personal protective equipment will need to be worn prior to starting any part of the experiment or setup and worn at all times. Individuals compliance with wearing PPE is extremely important. This protective attire helps to protect individuals from potential hazards such as spills, burns, splashes and much more that could harm one’s health. 2. As part of lab safety procedure and before beginning an experiment, what must you do in order to be prepared to conduct the steps of the experiment? (5 points) Prior to beginning any experiment, PPE should be applied, the procedure must be read twice thoroughly to ensure full understanding. If there is any question, do not proceed until clarification is received. The labels of all chemicals must be read. Location and review of the Safety Data Sheet. The selected area to perform the experiment should be a well-ventilated and a nonporous surface. All food, drinks or any items not directly related to the experiment should not within the selected area. The individual should have on close toed shoes with hair tied back. 3. Why are there increasing levels of biological containment procedures? What level of biological containment will be necessary to complete the experiments in this manual? (10 points) The levels of biological containment information provide pertinent information related to the characteristics of the microorganism or chemicals being used. When referring to characteristics, this includes but is not limited to the dangers, potential disease, limited knowledge, mode of transmission. If a microorganism is being dealt with in the lab of a disease with limited knowledge and no vaccine to date, this would be an example of a BSL-4. (Ebola is referenced on the eScience Labs resource, Biosafety Levels slide) The BSL required for the experiments to be completed fit within the BSL-1. The pathogen is unlikely to cause disease and present minimal potential hazard to personnel and the environment. (eScience Lab. Table 1: Biosafety Levels) 4. What document should you refer to if you have questions regarding the safety or disposal of a chemical? Where can this document be located? (10 points) The Safety Data Sheets can be referred to with any questions or concerns. Information on the proper handling and storage, accidental release and exposure, disposal, level of danger and actions that should be taken if one were to come in contact with a certain chemical can be found there. Students can find the SDS for every chemical in our lab kit on the eScience Lab website, https://esciencelabs.com/educators/msds. The information is also provided via the OSHA website. 5. Explain what the presence of dark to gray colored fuzzy colonies indicates on growth plates. (10 points) When the presence of dark to gray colored fuzzy colonies are observed on growth plates, this indicates a type of fungi or mold. Molds initially appear whitish gray with fuzzy edges and will gradually turn into the dark to gray color from the center on outwards. Fungal colonies appear “fuzzy” and bacterial colonies have a more smooth or rough appearance. Bacterial colonies also have a more defined margin, while fungal colonies are not quite as defined. (https://microbiologylearning.weebly.com/mycology.html) Experiment 1 Data Tables Table 2: Experiment 1 Colony Growth (10 points) Plate Condition 1 Hand without washing Hand with washing Yeast Deionized water 2 3 4 Growth Day 1 (0) Growth Day 2 (0) Growth Day 3 (+) Growth Day 4 (++) Growth Day 5 (+++) (0) (0) (+) (+) (++) (0) (0) (0) (0) (0) (0) (+) (0) (++) (+) Image 1 (10 points) Image: Insert an image of your growth plate #1 (when it has colonies present). Identifier: Include your identifier clearly visible and written on a piece of white paper, your name, student ID and the current, submitted date included in the image with the growth plate. (No credit for this lab will be provided unless both the growth plate and identifier are included in the same image and can be clearly identified). Experiment 1 Post-Lab Questions 6. What purpose did plating the yeast serve in this experiment? What purpose did the water serve? (10 points) Yeast serves as a positive control and water as a negative control. By using controls, this provides validity to the experiment being performed because if done properly the results are known beforehand. When a control is used, the individual can predict or will already have expectations on what will happen with each control. Therefore, this validates that the experiment was done properly. If the controls do not have the expected results, there was more than likely a contamination or error in the process of the experiment. When using the water, no result or no growth should appear. (https://whydoscientists.org/validating-experiments-controls/) 7. Which of the two hand treatments grew the most colonies? (10 points) The #1 plate by far grew the most colonies. There were multiple colonies of various sizes and colors. Primarily a creamy color, with 3 that are a goldish color and round-ish in shape. There are multiple that are smaller than the size the tip of a pen dot and a large group of colonies without defined margins where they are connected about the size of the nail on my pinky. (YUCK!) The rest of the colonies have defined edges and are almost a perfect circle. 8. What effect does hand washing have on the presence of bacteria? (10 points) While washing hands, initially the presence of bacteria increases because you just remove the transient bacteria, but you expose the resident microbes to the outer surface of your skin. However, with proper techniques while washing hands drastically reduces the amount of bacteria on your hands. The proper technique; use warm running water and soap, wash all surfaces, rub hands together for at least 15 seconds, rinse with warm water, hold fingers higher than elbows when drying. (https://microbeonline.com/handwashing-hygiene-wash-hands/) 9. Were there any colonies present on plate #2? What could account for the presence of these colonies? (10 points) The #2 plate (after handwashing) did have colonies present but not near as many as the #1 plate (no handwashing). There are several possibilities leading to the growth of bacteria on the #2 plate. My daughter (9) is also very interested in healthcare and has been learning some about Microbiology along with me. We decided to use her hands for this experiment. She very well could have not washed her hands properly, leading to the growth. As well as touching the faucet when turning the water off. She did very well at holding her hands up and not touching anything while we applied the glove to her hand.
