Abenaya Bio Formal Lab Report (1)

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“Attack of the Killer Fungus - Impact of Temperature Decrease on Arthrobotrys Oligospora ” April 1, 2022 Abenaya Murugathas 501084050 BLG144 Adisa Julien

1 Introduction This experiment was designed with the intention to increase knowledge of the scientific method in a practical manner (Sato, 2013). This includes the development of a hypothesis and experiment to accompany it. In this experiment, Arthrobotrys oligospora is used to trap Caenorhabditis elegans , and a T-test is used to measure the significance between data sets. A T-test helps scientists understand what is and what isn’t significant for their data. A T-test measures i f there is a significant difference between the means of two groups (Sato, 2013) . The P-value then measures if the difference may have just occurred by chance. In order for it to not be seen as random, the P-value must be lower than 0.05. A. oligospora are a part of the class Leotiomycetes, which is under the family Orbiliaceae (Sato, 2013). It can be found predominantly on compost, animal excrement, decomposing wood, and metal-polluted soil, and thus is considered one of the most abundant predacious fungi. In environments lacking nutrients, this fungi has an increased potency of catching prey. This then leads to trap formations. When they are not catching prey, the anatomy of the A. oligospora looks highly similar to that of a non-predacious species. The fungi would include features like hyphae and spore-forming apparatuses. With the help of stimuli like nematodes, the fungus is able to develop circular traps. In this experiment, the traps will be involved in capturing C. elegans . C. elegans are small, bacterivorous nematodes that have a rapid life cycle and a simple reproductive cycle (Sato, 2013). C. elegans , has an ideal growth temperature of 15–25 °C (Chen

et al., 2019). C. elegans are from the genus Caenorhabditis, which is a group of nematodes that reach up to 23 different species. C. elegans is unsegmented, cylindrical, and tapered at each end, which follows the body plan of a typical nematode (Sato, 2013). When taking a closer look at the body, it can be seen to be separated into two tubes by a pseudocoelom. There is an outer tube that holds the cuticle, muscle system, nervous system, hypodermis, and excretory system. There is also an inner tube holding the pharynx, reproductive system, and intestine. Homeostasis within the worm is maintained through internal hydrostatic pressure. C. elegans are also self fertilizing and are capable of producing 300 offspring. When they mate with males they can produce 1200 - 1400 offspring. A. oligospora is seen to have a peak growth rate at about 20°C to 25°C, and predatory height at 25°C to 28°C (Sato, 2013). In further studies, it was found that the fungus was significantly slower at capturing nematodes at lower temperatures. In the laboratory setting, C. elegans are considered the model organisms due to their short generation time, short life span, and small size, and homology to human genes (Sato, 2013). This study will explore if the change in temperature, particularly room temperature and a lower temperature of 16°C, reduces the trap formations in A. oligospora and thus increases the number of living C. elegans (Sato, 2013).

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2 Data The following results have been obtained following the experiment: Figure 1. The average ratio of surviving worms on the fungus plate and worms on the non-fungus plate after 6 hours. Control Plate Plate at 16°C Average 1.0818 1.0682 Standard Error 0.06932 0.07451 Average Survival Ratio 0.9874 T-test 0.132346 P-value 0.8998 Figure 2. The average ratio of surviving worms on the fungus plate and worms on the non-fungus plate after 24 hours. Control Plate Plate at 16°C Average 0.2398 0.3854 Standard Error 0.03495 0.1553 Average Survival Ratio 1.607 T-test -3.0311 P-value 0.02307

Figure 3. Visual comparison of worm survival between the two variable groups, room temperature vs 16°C. After performing the T-tests the results show that the plates at room temperature and the plates under 16°C for 6 hours have a significant difference of about 0.1323 and the P-value indicates that there is a high probability that the results are a coincidence. This is because the P-value, as seen in figure 1, is 0.8998 which is higher than 0.05. However, the T-test results for the plates at room temperature and the plates under 16°C for 24 hours has a significant difference of 0.0163 and a P-value of 0.02307, which indicates that there is a low chance of coincidence.

3 Discussion There were a couple of errors that happened when performing this experiment. For example, during the first attempt at viewing the worms under the microscope, there was some difficulty focussing and viewing the worms, which may have given us an inaccurate reading. There was another issue where it took too much time to read the sample of worms, and the sample had dried out. To correct this, we redid this portion of the experiment with a new sample to get more accurate results. The survival ratio between the two groups is found using the following method; Ratio = mean variable survival ratio / mean control survival ratio = 1.0682 / 1.0818 = 0.9874 After performing the calculations, it is found that the survival ratio of the plates after 6 hours id 0.9874, and the survival ration after 24 hours is 1.607. This shows how as time goes on, the ratio of worm survival on the plate at 16 °C is more than that of the control plates. This points in the direction that something is happening that is causing more worms to survive. On further examination of the data from this experiment, it can be seen that there are far more live worms on the plate that was under the 16°C variable than there were on the control plate, as seen in figure 3. Since it is known that C. elegans grows well at 16°C and at room temperature, it would be reasonable to say that the reason for and increase in worm survival at 16 °C is not due to the temperature change harming the worms. However, it is known that trap

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formations are negatively impacted by a temperature decrease such as the one in this experiment. This would have to mean that the trap transformations decreased at 16°C allowing for more worms to live, proving the hypothesis. The results in https://horizon.documentation.ird.fr/exl-doc/pleins_textes/fan/39963.pdf journal support the finding of this experiment. The journal also found that there was a decrease in trap formations in A. oligospora when exposed to lower temperatures. 4 Conclusion In this experiment, the aim was to explore the impacts of temperature on the Arthrobotrys oligospora and Caenorhabditis elegans . It was found that the temperature does have an impact on trap formation by decreasing it. There is a difference in worm survival ratio between the control and variable plates that lead to the acceptance of the hypothesis. A future study focusing on the incubation lengths may be beneficial in understanding how time plays a role in the number of worms surviving. 5 References Brian K. Sato, Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697 found in the JOURNAL OF MICROBIOLOGY & BIOLOGY EDUCATION, December 2013, p. 230-237 DOI: http://dx.doi.org/10.1128/jmbe.v14i2.612 Chen, Y.-L., Tao, J., Zhao, P.-J., Tang, W., Xu, J.-P., Zhang, K.-Q., & Zou, C.-G. (2019, June 13). Adiponectin receptor PAQR-2 signaling senses low temperature to promote C. elegans

longevity by regulating autophagy. Retrieved April 1, 2022, from https://www.nature.com/articles/s41467-019-10475-8#:~:text=For%20C.,leading%20to% 20a%20slowed%20aging . The influence of temperature, nutrition, light and the ... (n.d.). Retrieved March 28, 2022, from https://horizon.documentation.ird.fr/exl-doc/pleins_textes/fan/39963.pdf Appendix Figure 1. Observation of plates for ratio of surviving worms on fungus plate and worms on non-fungus plate after 6 hours. Control Plates Plates in 16°C Plate 1 1.158 0.931 Plate 2 0.961 1.185 Plate 3 1.032 0.818 Plate 4 1.132 1.042 Plate 5 1.126 1.365

Figure 1. Observation of plates for ratio of surviving worms on fungus plate and worms on non-fungus plate after 24 hours. Control plate Plates in 16°C Plate 1 0.112 0.357 Plate 2 0.198 0.387 Plate 3 0.324 0.327 Plate 4 0.214 0.421 Plate 5 0.351 0.435

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