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Sarah Serna 
Dr. Vetrone
INTD 100: Onco-Immunology
29 November 2017

The Effects of Vitamin B-12 on Caenorhabditis Elegans
Introduction: 
Our body produces a variety of different vitamins and hormones many of which are also subsequently taken as a method for controlling activity levels.  Vitamin B-12 is required for proper red blood cell formation, neurological function, and DNA synthesis.  Because Vitamin B12 assists in the converting of food to glucose that can be used as energy for the body, it has been thought that an increase in Vitamin B-12 could increase energy levels 1.  Vitamin B-12 is a nutrient in the vitamin B complex that the body needs in small amounts to function and stay healthy. It is found in liver, meat, eggs, poultry, shellfish, milk, and milk products 2.  Vitamin B-12 is water soluble and is needed daily in order for the human body to function properly. Vitamin B-12 is essential for DNA synthesis and for cellular energy production 3. Lack of Vitamin B-12 can lead to conditions such as anemia and neurological disorders 2.  

In order to conduct our experiment, we will be using Caenorhabditis Elegans (C. elegans) as our model organism.  Consisting of 959 somatic cells,  C. elegans show a very short life cycle and has the ability to change its locomotor behavior, reproductive rate, and lifespan 4. It has been estimated that about 42% of the human disease genes have an ortholog in the genome of C. elegans 5. Studies of cell and developmental biology that use C. elegans are greatly aided by the transparency of the animal, which allows researchers to examine development and changes due to mutations or altered environments 4.
 
Vitamin B-12 is essential for the growth of this worm, which contains two cobalamin-dependent enzymes, methylmalonyl-CoA mutase and methionine synthase 6.  C. elegans are likely to encounter both vitamin B12-replete and B-12-deficient diets in the wild because only a minority of bacterial species synthesize vitamin B-121. Previous studies have shown that the C. elegans that experience a deficiency in Vitamin B-12 exhibit a lower metabolic rate, as well as a slightly shorter life span5.  In our experiment, we will be focusing on how the addition B-12 to the food source affects the longevity and metabolism  of C. elegans to determine the effect of B-12 on the overall health of the organism

Materials and Methods: 

General Set Up: 
In order to conduct this experiment, we will need C. elegans, liquid vitamin B-12, and plates containing the E. coli OP50. Once the materials have been obtained, the C. elegans will be placed into three different groups, each initially containing 50 adult worms.  There will be a control group that is only given E. coli OP50 as its food source, a group with the B-12 at a 1x concentration in addition to the E. coli, and a final group with the B-12 at a 2x concentration.  For the first several days of the experiment, the adult worms from the original groups will be transferred onto new plates until they stop laying eggs.  Once they have finished laying eggs, the worms can remain on the same plate.  We will monitor the worms daily performing the different assessments, in order to determine the effect of Vitamin B-12 on the longevity, metabolism, and activity of the C. elegans.

Determine the Effect of Vitamin B-12 on Longevity of C. elegans
In order to test the effect of Vitamin B-12 on the lifespan of C. elegans, we will observe the three groups established in the methodology.  We will assess the groups for 20 days, removing any of the dead worms. After they are removed, we will recount the worms and record the total number in each group. 

Determine the Effect of Vitamin B-12 on the Metabolism of C. elegans
Assessing the metabolism of the C. elegans is an effective method to determine the overall health of the organism.  Using the previously established groups, we will then test the metabolism of the C. elegans by assessing their defecation rate.  To begin this assessment we must first obtain 4 day old C. elegans, that have each been exposed to their testing agent.  On the fourth day, we will test the defecation rate of six worms from each group by counting the number of pBoc contractions in each worm over a ten minute period. We will then take the average of each group.

Results: 
Determining the Effect of Vitamin B-12 on Longevity of C. elegans

On average, C. elegans live for between two and three weeks.  We wanted to test the effect of Vitamin B-12 on the length of the lifespan of C. elegans to determine if it could increase their longevity.  We discovered that at the end of the twenty days of the experiment, all three groups had a relatively similar number of alive C. elegans, with the control having an average of 36 worms remaining, the 1X group having an average of 36 worms remaining and the 2X group having an average of 37 worms remaining.  While the overall longevity of the worms was not greatly affected by the addition of Vitamin B-12, the worms at the 2X concentration tended to die off at a slower rate.  

Discussion:
Through our experiments, we discovered that Vitamin B-12 does not have a significant effect on the longevity of C. elegans While the exposure to Vitamin B-12 did not harm the organism or cause a decrease in lifespan, it did not prove to be statistically significant at extending the lifespan of C. elegans for long periods of time. For the group exposed to the 20% concentration if vitamin B-12, there was a period of time between days 7 and 16 where the exposure to B-12 appeared to help prolong the lifespan; however, those results did not remain throughout the extent of the project. For the defecation rate assessment, the worms exposed to the ten% concentration of Vitamin B-12 showed minimal changes from the control group; however, those exposed to the agent at a 20% concentration showed an increased rate of metabolism. The difference in the defecation rate of the worms can be linked to an overall increase in health of the worms that were exposed to a 20% concentration of Vitamin B-12.  Previous studies concerning C. elegans and Vitamin B-12 have shown that when in deficiency of vitamin B-12, the life span can be shorter; however, they don’t discuss the effects of adding excess Vitamin B-12 to the diets of C. elegans 5.  Vitamin B12 deficiency has been linked to developmental disorders, metabolic abnormalities, and neuropathy; however, the mechanisms involved remain poorly understood 7. Overall, we were able to conclude that additional Vitamin B-12 does not significantly impact the life span of C. elegans; however, without the proper amount of Vitamin B-12, it can be harmful to the organism.  

Conclusion:
Our experiment allowed us to determine the effects of Vitamin B-12 on the longevity and overall health of the organism. Through our research we determined the effects of vitamin B-12 on the metabolism and longevity of C. elegans. Because vitamin B-12 occurs naturally in our bodies, determining how the C. elegans react when their natural supply is supplemented could provide a better explanation on the effects of  vitamin B-12 on humans. While we can only truly determine the effects of Vitamin B-12  on C. elegans, because of their similarity to the human genome, our research could provide a variety of people with a possible natural solution to increase their health that would prevent the need for chemically enhanced treatments.  

Works Cited

California Healthcare Foundation/American Geriatrics Society Panel in Improving Care for Elders with Diabetes, Care. “Guidelines for Improving the Care of the Older Person with Diabetes Mellitus.”Journal of the American Geriatrics Society, Blackwell Science Inc, 29 May 2003, onlinelibrary.wiley.com/doi/10.1046/j.1532-5415.51.5s.1.x/full.
“Vitamin B12 – National Library of Medicine – PubMed Health.” National Center for Biotechnology Information, U.S. National Library of Medicine, www.ncbi.nlm.nih.gov/pubmedhealth/PMHT0022013/.
O’Leary, Fiona, and Samir Samman. “Vitamin B12 in Health and Disease.” Nutrients, Molecular Diversity Preservation International, Mar. 2010, www.ncbi.nlm.nih.gov/pmc/articles/PMC3257642/.
“Vitamin B12 – National Library of Medicine – PubMed Health.” National Center for Biotechnology Information, U.S. National Library of Medicine, www.ncbi.nlm.nih.gov/pubmedhealth/PMHT0022013/.
Maria Markaki M. Modeling human diseases in Caenorhabditis elegans. 2010. http://materiais.dbio.uevora.pt/MA/Modeling_human_diseases_in_Caenorhabditis_elegans.pdf
Lee , Seung-Jae. Glucose Shortens the Life Span of C. Elegans by Downregulating DAF-16/FOXO Activity and Aquaporin Gene Expression. 4 Nov. 2009, www.cell.com/cell-metabolism/fulltext/S1550-4131(09)00302-7?_returnURL=http%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS1550413109003027%3Fshowall%3Dtrue.
Bito, T, et al. “Vitamin B12 Deficiency in Caenorhabditis Elegans Results in Loss of Fertility, Extended Life Cycle, and Reduced Lifespan.” FEBS Open Bio., U.S. National Library of Medicine, 1 Feb. 2013, www.ncbi.nlm.nih.gov/pubmed/23772381.