Proposal to Test the Impact of the Genus Ulva on B12 Serum Levels in Vegans

by Renata Meyer-Pflug

Kristine M. Schweitzer

Gloria Maria Lobato

Monick David

Cabalamin a.k.a Vitamin B12• Cobalamin is:

– Water soluble– Found Primarily in animal sources– Extremely important for normal functioning of the

brain and nervous system• Deficiency in Cobalamin causes

– Anemia• Condition in which the body does not have enough

healthy red blood cells– Megaloblastic anemia

• Anemia with larger-than-normal red blood cells– Peripheral neuropathy

• Condition that involves a disruption of the neuro-electrical synapses from the brain and spinal cord to the muscles in the limbs and other parts of the body

Ulva spp. a.k.a. Sea Lettuce

• Ulva spp. stands for Several Species• Species include

– Ulva Lactuca , commonly known as sea lettuce

– Enteromorpha, commonly known as green nori

• Ulva spp.– Is popular in cuisine of Scandinavia, Great

Britain, Ireland, China, and Japan– Contains high levels of carbohydrates,

minerals, vitamins and trace minerals– Contains a very high content of vitamin B12

Ulva spp. a.k.a. Sea Lettuce

Nutritional Value of Edible Seaweeds by MacArtain et al., 2007

Problem Statement

• Certain populations are at risk for B12 deficiency– The elderly– Individuals with gastric diseases– Vegans

• Although vegan diets promote good health, the risk of deficiency is still prevalent

• Vegans wanting to avoid the problems vitamin B12 deficiency causes are forced to take synthetic supplements

• Currently there are no studies on whether or not the vitamin B12 present on the Ulva spp. is bioavailable to the humans.

Hypothesis

“Two hypotheses emerge after the review of the

literature. One is that vitamin B12 is existent in

the Ulva spp. algae and is bioavailable to the

human body. Second, is that Ulva spp. can be

manufactured into supplements for the purpose

of preventing the very prevalent cobalamin

deficiency in vegans.”

Literature Review

• Little research has been done in the larger field of sea-vegetables

• Some studies confirm the existence of vitamin B12 in certain strains of marine algae

• Some research results are still inconclusive due to the variation of the vitamin content in these foods.

• Caution is suggested in recommending them as a source of reliable vitamin B12

Nutritional Value of Edible Seaweeds

• Studies on vitamin B12 levels of vegetarians reveals that substantial amounts of B12 were found in purple and green algae

• This statement is supported in the research on “Isolation of Folate-producing Microalgae” done in Japan

• An investigation done on the “Nutritional Value of Edible Seaweed” shows Ulva spp. as having the highest content out of all tested

• Based on this list, Ulva spp. It was determined that Ulva can provide B12 vitamin in excess of the RDA

Bioavailability of Algae in Humans

• Bioavailability of vitamins and minerals is often an issue when dealing with vegetables and fruits.

• All the cobalamins that cannot be converted into the active form of vitamin B12 are known as ‘B12 analogues

• Analogues are biologically inactive molecules, which cannot be utilized by the human body”.

• In the same article the researcher affirms that “algae and cyano bacteria have long been considered to be a reliable natural sources of vitamin B12”.

Bioavailability cont’d

• According to a study conducted by the Department of Bimolecular Sciences, University of Urbino in Italy, many analytical methods cannot distinguish these analogues from true vitamin B12.

• As a consequence, the content of true vitamin B12 in food, as well as its blood concentration can be overestimated.

• To dispute that, a study done by the Institute of Nutritional Sciences in Germany maintains that this particular algae, based on its high content of true vitamin B12, contains active B12 and can be recommended for vegetarians as a dietary source of B12.

Literature Review-Summary

• Researchers utilized food models, online software, clinical trials and trained professionals for collecting accurate data on the content of vitamin B12 in seaweed and its bioavailability to the human body.

• All the studies done on the vegan population agree on the deficiency and need for a natural source as a supplement as opposed to the ones produced by bacterial synthesis done in laboratory.

• To our knowledge, the studies that we have reviewed have shown that cobalamin from algae is mainly comprised of analogues without true biological activity in humans.

Research Objectives

• Because the literature review has not identified a study that tests bioavailability of vitamin B12 in vegans there is a need for further research

• Our research aims to – Identify 100 vegans meeting our eligibility

criteria– Administer Ulva spp. to the vegans– Test vitamin B12 Serum Levels– Record results– Prove that a relationship exists between

• Independent variable-Ulva spp.• Dependent Variable-Serum Vitamin B12 levels

Research Design• Single Blind Experimental Study Design• One-hundred participants selected• Separated into two groups using a table of random

numbers– Group 1, Control Group receives B12 supplement– Group 2, Experimental Group receives a specially designed

vegan seaweed corncakes

• Diving, cleaning, drying, and corncakes preparation will be in accordance of methods used by a study on the iron bioavailability of Ulva spp.

• Corncakes will be delivered to the participants homes

• Corncakes must be eaten by participants three times daily for 30 days

Vegan Corncakes • 1 cup yellow cornmeal• 3/4 cup unbleached flour• 1 1/2 cups water or vegetable broth• 1/2 onion, finely chopped• 1/2 tsp sea salt• 4 grams Ulva Lactuca• Mix cornmeal and flour together in a large bowl.• Bring water or broth to a boil, then pour over

cornmeal mixture. Stir together and set aside. (Should be thick.)

• Saute onion in vegetable oil until translucent. Season with salt, cook until onions are soft and lightly brown, add Ulva Lactuca, then fold into cornmeal mixture.

• Cover the bottom of a skillet with oil and heat while you form the dough into patties

Research Design Continued

• Participants will fill out a food intake form daily• The intake form will be evaluated by the same

trained technician• Data will be input into a specialized software

package • Blood serum levels will also be analyzed using

specialized software• The methodology used to monitor food intake and

vitamin B12 serum levels was also used to check the vitamin B12 status of elderly Koreans by Kwak et al. in a 2010 study

Population and Sampling

• Eligibility Criteria– Strict Vegan (observed with pretesting)– Male or female between 14-50– Overall good health– Lives within a 25 mile radius of Montclair State

University– Deficient in Vitamin B12 (observed with

pretesting)– Currently do not take a vitamin B12

supplement• Participants will be recruited from any of the seven

vegan restaurants chosen until a quota of 100 is met.

Rationale for the Study Design

• Allows manipulation of variables• Establishes causality between variables

– Algae intake– Vitamin B12 serum levels

• Replication would be possible• Requires a smaller sample size• Represents diverse characteristics of the

population

Data Collection

• Two Data Collection Methods– Food intake questionnaire

• Filled out by participant• Evaluated and input into a software

program by the same trained technician• Pretesting will rule out ineligible vegans

– Blood Tests• Collected at 3 points, Day 1, 15, and 30• Transported to the laboratory in cold ice

boxes• Uses the Radioimmunoassay equipment

to measure serum levels

Daily Food Intake of Subjects

DRAFT

Research Design-Summary• Our experimental research design takes the best

methods of administering seaweed and testing serum vitamin B12 levels

• Other studies similarly use experimental methods to test the effect of algae in humans, but some utilize cross sectional analyses– observation of all of a population, or a

representative subset, at one specific point in time.

• Limitations of our experiment are confounding factors such as poor absorption in participants and unknowing consumption of B12 fortified products

Summary

• A successful experiment would promote advancement of science and knowledge

• Vegans and individuals who are looking for a natural alternative for vitamin B12 supplementation would benefit from this study