Nicodemus Wilderness Project -- Global Registry of Apprentice Ecologists

AN ANALYSIS OF NATURAL AND AGRICULTURAL SETTINGS ON THE CELLULAR RESPIRATION RATES OF SUBTERRESTRIAL LIFE Abstract The rampant use of inorganic fertilizers has posed a threat to subterrestrial life and the agricultural sector at large. Such inorganic compounds impact subterrestrial life, and the inhibition of cellular respiration is a potential mechanism by which it exerts these effects. This notion, namely the use of fertilizers as degrading subterrestrial life, inspired me to analyze the effect of Miracle-GroÂ® All Purpose Plant Food on the cellular respiration rates of Eisenia fetida as my Apprentice Ecologist project. Our hypothesis speculated that subterrestrial life subjected to Miracle-GroÂ® All Purpose Plant Food experienced fertilizer-induced stress, decreasing the respiration rate of E. fetida and other subterrestrial life. Here, standardized gas collection procedures were established, and CO2 levels were observed to examine the underlying factors proliferating and reducing respiration rates in such organisms. Two collection chambers simulating agricultural and natural environments were established, and CO2 gas collection was recorded. Average respiration rates were recorded as 17.47 and 32.93 ppm/sec, and standard error of the mean (SEM) was 22.85 and 1.40 ppm/min in the simulated agricultural and natural environments respectively. Our findings provide strong evidence that short-term exposure to Miracle-GroÂ® All Purpose Plant Food has no significant difference on the respiration rate of E. fetida and other subterrestrial organisms. However, we assert that the presence of Miracle-GroÂ® fertilizer induces fluctuating CO2 levels, and we speculate that such compounds likely inhibit cellular respiration mechanisms long-term. We found that Miracle-GroÂ® fertilizer treatment can induce volatile respiration rates, ranging from 44.6 to -27.6 ppm/sec. Not only do these findings suggest inhibited decomposer activity when exposed to various fertilizers, but they also point to the underlying lethality of pesticides and inorganic fertilizers on subterrestrial life. This potential, stark decrease in decomposer biodiversity could deteriorate the O and A horizons altogether, thereby leading to reduced crop yields in the agricultural sector. Moreover, previous studies have linked such soil degradation with drastic reductions in the soil's water holding capacity and nutrient-recycling capabilities. Thus, while Miracle-GroÂ® All Purpose Plant Food may yield increased profitability temporarily, the present study suggests a drastic reduction in organism biodiversity and nutrient availability long-term. Keywords: cellular respiration, Miracle-GroÂ®, fertilizer, respiration rate, CO2 collection, gas collection procedure, agricultural environments, Eisenia fetida 1

Introduction Recent fertilizer treatment studies provide strong evidence supporting the notion that pesticides and inorganic fertilizers contribute to benefits in the agricultural sector, such as increased crop yields (Atkar, et al., 2009). Yet other studies have indicated that these same chemicals, among them synthetic insecticides and plant growth regulators, exert wide ranging consequences to non-target species, namely "humans, animals, and higher plants" (Atkar, et al., 2009). One such organism that is adversely impacted by these non-selective pesticides is the red wiggler Eisenia fetida, leading to a substantial reduction in their population and thus endangering the subterrestrial ecosystem at large (Miglani and Bisht, 2020). Moreover, exposure to phosphorus-containing plant-growth regulators, inorganic fertilizers, and synthetic pesticides has been linked to reduced energy production in plants but lacks experimentation in heterotrophic organisms. (Letham, 1969). Thus, central to understanding the threats posed by these synthetic pesticides is to understand the cellular respiration rates not only in plants but also in E. fetida and other subterrestrial life. This study observes the effect of the synthetic fertilizer Miracle-GroÂ® on these living forms by determining their rate of cellular respiration. Cellular respiration (hereafter respiration) is a ubiquitous mechanism that converts the potential energy within the bonds of glucose into a widely expendable form of energy termed adenosine triphosphate (ATP). It is illustrated by the equation C6H12O6 + 6O2 ?' 6CO2 + 6H2O + ATP (Bridges, 2021). This catabolic mechanism is essential to sustaining life, as it provides cells with widely-accessible energy driving its various metabolic processes. 2

Materials ?--? E. fetida, commonly referred to as ?--? the "Red Wiggler," 9 worms ?--? ?--? Glass Beads, 25 ?--? ?--? Glass Beaker ?--? Graduated Cylinder ?--? ?--? Miracle-GroÂ® All Purpose Plant Food, 0.33 g ?--? ?--? Miracle-GroÂ® Potting Mix, 450 mL Styrofoam cup Tap Water, 88.8 mL VernierTM GO Direct Gas Collection Chamber, with graduations VernierTM GO Direct CO2 Gas Sensor Vernier Graphical AnalysisTM Web-Based Application ?--? Scoopula ?--? Standard Electronic Balance ?--? Weigh boat Procedures Heterotrophs and Gas Collection Procedure This study received approval from the Department of Biology, a subdivision of the Department of Sciences in Stratford Academy. E. fetida, were housed in 23 ? 2Â° Celsius in regular atmospheric conditions. In addition, the gas collection procedure was conducted in identical, room temperature conditions under continual light. The procedure is as follows: 1. Specimens and the materials needed for their respective treatment were obtained, adequately measured, and placed in the horizontally-oriented gas collection chamber. 2. A Vernier GO Direct CO 2 Gas Sensor was superficially placed within the chamber and connected via Bluetooth. The Vernier Graphical Analysis application was opened, and on-device instructions were correctly executed. 3

3. Specimens were granted a one minute acclimation period in the gas collection chamber, and initial CO2 conditions were recorded to the nearest 0.01 ppm (see Tables 1 and 2) 4. CO2 readings were subsequently recorded at one minute intervals over a five minute duration to the nearest 0.01 ppm (see Tables 1 and 2) 5. Gas collection chambers were shaken after the five minute duration, effectively removing any previous, gaseous contaminants. The procedure was repeated and readings were recorded as necessary. CO2 Collection in Treated and Untreated Subterrestrial Environments Respectively The standardized gas collection procedure was derived from the section entitled "Heterotrophs and Gas Collection Procedure," and CO2 levels were recorded in E. fetida and other subterrestrial life. In brief, E. fetida worms were placed in typical subterrestrial conditions or exposed to an H2O-fertilizer solution in the control and experimental groups respectively. E. fetida were randomly assigned (n = 3 per group), and CO2 levels were recorded in adherence to steps three and four of the general CO2 collection procedure. Thus, E. fetida effectively modeled natural and agricultural settings, with procedure conducted as follows: 1. 75 mL of Miracle-GroÂ® Potting Mix was proportioned into two identical collection chambers, modeling the control and experimental groups respectively. 2. Two measurements of 14.8 mL H2O were apportioned into two graduated cylinders, one of which was directly added to the collection chamber modeling the control environment. 3. Dimensional analysis was conducted in adherence to the listed one tablespoon of Miracle-GroÂ® All Purpose Plant Food per one gallon of water, yielding 0.11 grams of fertilizer per 14.8 mL of water. The target quantity was subsequently measured using a scoopula and electronic balance, and the fertilizer was mixed thoroughly with the remaining 14.8 mL H2O. 4. The remaining apportionment of H2O was allocated to the gas collection chamber modeling the experimental group, and a Vernier GO Direct Gas Sensor was prepared via the general recording procedure established by step two (see Heterotrophs and Gas Collection Procedure). 5. Three worms were randomly apportioned to the control and experimental groups respectively (n = 3 per group), using different worms for each repeated trial. 6. Steps three, four, and five in the general gas collection procedure were observed, and three trials were conducted per group. 4

Discussion The rampant use of pesticides and inorganic fertilizers has posed a pervasive threat to the future of E. fetida, the agricultural sector, and all of subterrestrial life. The turn of the 21st century has seen widespread implementation of these non-specific chemicals, thereby increasing the incidence of mortality in E. fetida, and, ultimately, the destruction of subterrestrial life (Miglani and Bisht, 2020). This stark decrease in decomposer biodiversity could deteriorate the O and A horizons altogether, drastically degrading the soil, contributing to desertification (i.e., a reduction in water holding capacity), and reducing crop yields in the agricultural sector altogether (Eisenhauer, et. al., 2018 and Herrmann, et. al., 2018). The objective of this study was to determine the impact of one such compound, Miracle-GroÂ® All Purpose Plant Food, and its linkage to the cellular respiration pathway at large. Cellular respiration catabolizes chemical energy stored within the bonds of glucose into the widely expendable, "energy currency of the cell" termed ATP. Thus, cellular respiration is a ubiquitous mechanism fueling metabolic pathways and thus sustaining all life on earth (Bridges, 2021). Here, by simulating agricultural and natural environments in a gas collection chamber, we found that short-term exposure to Miracle-GroÂ® fertilizer has no observable effect on the respiration rate of E. fetida and other subterrestrial life. In this study, we speculate that long-term pesticide and chemical exposure could lead to the inhibition of the respiration pathway, jeopardizing the future of E. fetida and subterrestrial organisms at large. We provide compelling evidence that the presence of Miracle-GroÂ® fertilizer induces fluctuating CO2 levels in subterrestrial life. Although the medians of the fertilizer-treated and untreated categories differed only trivally, by 0.4 ppm/sec, a marginal, 15.06 ppm/sec difference was observed in the ranges of the two groups. With a range of 15.93 ppm/sec in the 5

fertilizer-treated group alone, this data provides strong evidence that inorganic fertilizers and pesticides can inhibit the cellular respiration pathway long-term. Despite the limitation that duration and repeated trials poses on this study, stress as a causative factor for fluctuating CO2 levels in fertilizer-treated conditions cannot be dismissed. Short-term, fertilizer-induced stress likely affected not only E. fetida activity but also caused fluctuating respiration rates in subterrestrial life. This study provides strong evidence linking Miracle-GroÂ® fertilizer treatment with volatile respiration rates, ranging from 44.6 to -27.6 ppm/sec in Trials 2 and 3 respectively. Additionally, the simulated agricultural environment reported increased E. fetida activity likely linked to fertilizer-induced stress. Our findings reported here indicate that volatile respiration rates are directly correlated with increased E. fetida activity in the simulated agricultural environment. Our results refute the hypothesis that fertilizer-induced stress has negative effects on respiration rates in subterrestrial life but remains inconclusive over longer durations (n > 5 minutes). The shorter duration (n = 5 minutes) at which E. fetida were inputted into their fertilizer treated and non-fertilizer treated environments is a likely cause for the absence of a significant difference in the respiration rates of treated and untreated subterrestrial life. Thus, our results support the absence of a significant difference at shorter durations but remain inconclusive long-term. In addition, residue from both simulated and unsimulated environments could have come into contact with the VernierTM GO Direct CO2 Gas Sensor, yielding undesired CO2 readings that could account for the volatility in the treatment-added group. Our findings shown here have indicated that notable increases in activity in the presence of fertilizer treatment caused certain E. fetida to tamper with the gas sensor, potentially yielding altered or inaccurate readings altogether. Nonetheless, the present study indicates that fertilizer-treated conditions have no 6

short-term consequences to the respiration rates of subterrestrial life, but we speculate that fluctuating respiration rates may inhibit the cellular respiration pathway long term. Pesticides and inorganic fertilizers are one of the most pervasive problems confronting subterrestrial organisms and the agricultural sector at large. The present study serves to isolate the biological process directly affected in subterrestrial life. Here, in modeling agricultural and natural environments, we speculate that fertilizer-treated environments likely affect the cellular respiration pathway in E. fetida and other subterrestrial life long-term. Thus, in isolating the cellular respiration mechanism as a probable factor in eradicating subterrestrial life, the present experimentation can expedite the development of a treatment mending the respiration pathway when exposed to these inorganic compounds, or it can encourage a switch to their organic counterparts altogether. Thus, the present Apprentice Ecologist Project highlights the benefits of organic fertilizers on subterrestrial life, as they exhibit many of the advantages of their inorganic counterparts, while still maintaining soil biodiversity. 7

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