2. 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

3. 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

4. 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

5. Ulva spp. a.k.a. Sea Lettuce
Nutritional Value of Edible Seaweeds by MacArtain et al., 2007

6. 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.

7. 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.”

8. 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

9. 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

10. 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”.

11. 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.

12. 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.

13. 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

14. 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

15. 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

16. 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

17. 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.

18. 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

19. 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

20. Daily Food Intake of Subjects

21. 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

22. 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