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Fall 2009 Biology 3B Paper DISCUSSION Though the results of no statistically significant difference, differed from previous findings by Gee and So (2009), the overall inactivation at 60s of irradiation reflected those found by Latimer and Matsen(1977). The difference in findings between this experiment and that of Gee and So(2008)are most likely attributed to greater number of intervals ran as well as different statistical analysis. Difference in findings may also be due to Gee and So having used a standard t-test as opposed the more accurate ANOVA. Further investigation into the cellular effects of the exposures to 100°C water bath and microwave irradiation is of interest. LITERATURE CITED Celadroni F., Giannessi F., Ghelardi E., Longo I., Tosoratti N., Baggiani A., Salvetti S., and Senesi S. 2004. Effect of microwave radiation on Bacillus subtilis spores. Applied and Environmental Microbiology, 97: 1220- 1227 Chipley J.R., Dreyfuss M.S. (1980) Comparison of effects of sublethal microwave radiation and conventional heating on the metabolic activity of Staphylococcus aureus. Applied and Environmental Microbiology, 39: 13-16 Gee B. and So J. (2009) The Comparative Efficiency of Sterilization by conventional Heating And Microwave Irradiation On E. coli. Saddleback Journal of Biology 7,119 Górny R., Harkawy A., Kasznia-Kocot J., Lis D., Łudzeń-Izbińska B., Mainelis G., Marzec S., Niesler A., Siwińska E., Wlazło A.(2007) Viability of fungal and Actinomycetal spores after microwave radiation of building materials. Ann agric Environ Med 14: 313-324 Latimer J, Matsen M. (1977) Microwave oven irradiation as a method for bacterial decontamination in a clinical microbiology laboratory. J Clinical Microbiology, 6: 340-342 Park H-D., Rhee I-K, Woo I-S. (2000) Differential damage in bacterial cells by microwave radiation on the basis of cell wall structure. Applied and Environmental Microbiology, 66: 2243-2247 The Effect Temperature has on the Aerobic Metabolism of Cold-Water Adapted and Warm-Water Adapted Fish Krystina Jarema Department of Biological Sciences Saddleback College Mission Viejo, California, 92692. The metabolic rate of fish should be optimum in the adaptive temperature of the fish. Testing the validity of this assumption, tilapia (Oreochromis niloticu; warm-adapted fish) was compared to salmon (Oncorhynchus nerka; cold-adapted fish) using succinic acid and methylene blue as indicators of aerobic metabolic activity. Each sample was chopped and homogenized with sodium phosphate solution. Ten mL of each homogenized sample was iced in labeled test tubes with three drops of methylene blue and 0.5 mL of succinic acid and placed into a water baths set to 0 o , 7 o , 15 o , 22 o , and 30 o C. The faster the sample metabolized, the faster the methylene blue was reduced to a colorless form due to the oxidation reduction process. The time it took for half of the methylene blue to be oxidized from the each sample was recorded and compared to the species as a whole, and the temperature of the bath they resided in. The average combined times for tilapia (37.2 ± 5.7 minutes) was not significantly faster (p=0.1652 two-tailed unpaired t-test) than the mean times of the salmon (69.8 ± 8.7 min). Thus neither fish was able to adapt to a greater range of temperatures and metabolize at a significantly faster rate than the other within in different temperature. 147 Saddleback Journal of Biology Spring 2010
Fall 2009 Biology 3B Paper Introduction The adaptive properties of fish allow them to metabolize optimally at a certain temperature range. Tilapia is native to 20-30°, and salmon inhabit areas that range in temperatures from 0-15°, according Irons III and Oswood (1991). It is logical that the warm water adapted fish metabolize better in high temperature water than a cold adapted fish would. The body temperature of fish remains close to that of the water they reside in and their capability to adapt to marginal thermal change makes fish excellent ectothermic organisms in which to examine temperature responses (Guderley, 2004). Since tilapia has a higher temperature tolerance, its metabolism is able to function at high temperatures and not bodily over-heat them; the higher the temperature set point, the greater the endurance of high-rate aerobic activity at high ambient temperatures (Heinrich 1977). When distinguishing a fish as being either warm or cold water adapted, the fish are labeled by the temperature ranges that they metabolize optimally at. Tilapia resides in tropical waters where as salmon reside in arctic to temperate winter waters and as such the fish’s internal body temperature have adapted to account from their habits. Enzymes act primarily as catalysts, or as controlling or regulatory elements within an organism. Enzymes need to be able to fluxuate in order to initiate changes during strategic positions in metabolism. If enzymes were non-adaptive, the organism would die due to its inability to metabolize or regulate itself when a change in the body system occurs (Hochachka and Somero, 1984). Succinc Acid is an enzyme that is required in the Citric Acid Cycle for aerobic metabolism to occur. By using this enzyme, a metabolic reaction is triggered within the samples, which in term allows them to undergo aerobic metabolic activity after death. As SDH triggers the intermediate to change from succinate to fumarate releasing FADH 2 which intern is used for oxygen reduction. When that oxygen reduction occurs, a reaction with the methylene blue changes it to its colorless form showing the metabolic progression of the sample. Our objective was to determine if warm water adaptive fish metabolize better that cold adaptive fish in an incubator set to 37 o using succinic acid and methylene blue as indicators of aerobic metabolism. I hypothesized that the there would be a significant difference in the rate it took for oxidation-reduction to change the methylene blue to its colorless form within the samples. Materials and Method Five samples of tilapia and five samples of salmon were used in this study. The salmon was obtained from Jon’s Fish market, Dana Point California on 20, 21, 22 nd of November 2009, and the tilapia was purchased from Vons, Laguna Niguel California on the 22 nd of November 2009. All measurements were made on 22 nd of November 2009. The samples were chopped into pieces and weighted out to ten grams (g). The samples were then frozen at -80 oC each wrapped in both plastic and foil individually and labeled. Ten g of each sample was thawed out to room temperature; each sample was homogenized together with 25 mL of sodium phosphate solution individually; The sodium phosphate (Solution C) is comprised of [306 mL sodium phosphate monohydrate (Solution A) (NaH 2 Po 4 +H 2 O; 31.73 g to 1000 mL) + 255 mL of Sodium phosphate Dibasic (Solution B) (NaHPo4 + H2O; 53.6 g to 1000 mL) + 600 mL of Deionized Water (DI)]; the sodium phosphate solution acts as the buffer for this experiment. Ten mL of the each homogenized sample were placed in labeled test tubes and placed back into the freezer set at -20 oc . the samples were thawed as five water baths were set to specific temperatures (0 o , 7 o , 15 o , 22 o , 30 o ). Three drops of methylene blue was added to each sample followed by .5 mL of succinic acid (59g/500mL x 10 mL Solution C, a total of 1.18g of succinic acid was added to 10 mL of Solution C and heated until dissolved). A glass rod was used to mix the samples until the methylene blue fully permeated the samples. The two test tubes, one of each fish species, were placed in each of the water bathes. The samples were continuously checked on at fifteen-minute intervals until visible signs of oxidation in the samples were noticed. Then the samples were checked every five minutes until the half the samples were clear. The oxidation reduction that is under gone during aerobic metabolism and how quickly it is being done is what is being measured when the speed in which it took for each sample to reduce the entire methylene blue within sample to its colorless form is recorded. All data was transferred to MS Excel (Microsoft Corporation, Redmond, Washington) where all further statistical manipulations were performed. Results The average combined times of warm-water adapted tilapia in this study was 37.2 ± 5.7 minutes. The average combined times of cold-water adapted salmon in this study was 69.8 ± 8.7 minutes. A twotailed unpaired t-test revealed that the mean combined times of the two fish were not significantly different (p=0.165212546). 148 Saddleback Journal of Biology Spring 2010
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