the effect of plant-based prebiotics on iron

The Effects of Plant-Based Prebiotics on Iron Bioavailability, Beneficial Bacteria

Populations, and Intestinal Functionality through Intra-Amniotic Administration

Jessica Cho

Review of Literature

Research Question

Hypothesis Methods Results/Discussion Bibliography

Micronutrient Malnutrition (Miller and Welch, 2013)

• Diet lacking in adequate vitamins and minerals • 1/3-1/2 world population

www.fertilizer.org


Research Question Hypothesis Methods Results/Discussion Bibliography

http://www.ifpri.org/sites/default/files/ghi14tab31lg.jpg

Causes of Deficiency


Research Question


Prebiotics• Non-digestible carbohydrates/fibers• Serve as raw material for gut bacteria and other microbes to utilize

for energy• Stimulates the growth of specific species of gut bacteria

(bifidobacteria, lactobacilli, etc.)

lactobacilliBifidobacteria

(Tako et al., 2014), (Schrezenmeir and Vrese, 2001)


Research Question


Beneficial Effects of Prebiotics (Tako et. al, 2013), (Cummings and Macfarlane, 2002), (Gibson and Roberfroid, 2010)

Decrease pro-inflammatory immune factors

Increase uptake of certain minerals

such as Fe and Zn

Increase production of short chain fatty acids

Increase “good” bacteria in the gut

Improved bowel regularity

Review of Literature Research Question


Probiotics-help.com

http://img.sparknotes.com/figures/9/92fefe6f9160757a6207a120b3637c38/figure4.gif

Microvilli cover surface of epithelium (tissue that lines cavities of surfaces of blood vessels and organs), -enzymes that allow for the absorption of digested nutrients

Research Question


Research Question


Is the functionality of the brush border membrane, the abundance of bacteria populations, or iron bioavailability altered with the

introduction of various prebiotics?

Hypotheses


Research Question


H0: The intra-amniotic administration of the prebiotics will have no effect on iron bioavailability, gut bacterial growth, or the brush border membrane.

H1: The intra-amniotic administration of each prebiotic group will increase beneficial bacterial growth.

H2: The intra-amniotic administration of each prebiotic group will have an effect on the functionality of the brush border membrane.


Research Question


Broiler Chicken Model (Tako et al., 2010)

• Fast growing time• Short incubation time• Sensitive for dietary mineral deficiencies • >85% homology between gene sequences of human and chicken

intestinal DMT-1, DcytB, and Ferroportin

http://www.marketingandtechnology.com/repository/webstories/webst15985.png


Research Question


http://www.poultryhub.org/wp-content/uploads/2012/07/chick_embryo_development1.jpg

Development of the Broiler Chicken

Intra-amniotic Administration Procedure (in ovo feeding technique)


Research Question


1) 2) 3)

4) 5) 6)


Research Question


Treatment GroupsGroup # Treatment Vol Medium Staple Crop1 2% Sorbitol 20mL Saline Lentils (Africa,

India)2 2% Mannitol 20mL Saline Lentils (Africa,

India)3 2% Raffinose 20mL Saline Beans, peas,

lentils(Africa, India)

4 0.5% Stachyose 15mL Saline Beans, peas, lentils(Africa, India, S America)

5 2% Curcumin 20mL Oil Curcuma longa turmeric (India and Pakistan)

6 Oil7 Saline8 None


Research Question


Procedures1) Euthanization of 21 day old broiler chicks

• CO2 exposure2) Draw blood

• Determination of hemoglobin concentrations spectrophotometrically

3) Collection of small intestine (Gene Expression Analysis)• RNA extraction using Rneasy Mini Kit• RT-PCR used for amplification• Gel Electrophoresis—band densities measure relative

abundance4) Collection of cecum (Bacterial Analysis)

• DNA extraction• PCR used for amplification• Gel electrophoresis—band densities

5) Statistical Analysis• MANOVA test using JMP software• P<0.05

Intestinal (cecum) Content Bacterial Genera

Analysis Results

Positive: ↑ beneficial bacteria

Positive: ↓ harmful bacteria

BBM Functionality Reflected by Enzyme Gene Expression

Results

Sucraseisomaltase

Amino Peptidase

1

2

3

4

1

Suggests an increased absorbable/digestive effect on protein at the BBM

2

Suggests effect on absorbable/ digestive functionality of BBM

3

Suggests increased absorption of sucrose and maltose into intestinal epithelial cells

4

Lower expression Increased iron absorption


Research Question


intestinal beneficial bacteria

SCFA production decrease in pH

Promotes production of ferric iron (Fe3+) to ferrous

iron (Fe2+)

Increase of soluble iron in

intestine

Increase iron absorption

improved iron status


Research Question


Prebiotic Treatment Groups

Curcumin Sorbitol Mannitol RaffinoseStachyose

Genes and Enzymes at BBMDMT-1, AP, SI, ATPase

Suggests Stimulation of the BBM Functionality

Possibly enhances its absorptive/ digestive functionality


Research Question


Future Research

Sorbitol

Mannitol

Raffinose

Stachyose

Curcumin

Biofortification: development of micronutrient-dense staple crops using traditional breeding practices and modern biotechnology

Curcuma longa Turmeric India and Pakistan

lentil

lentil Fresh mushrooms

Beans, peas, lentilsAfrica India

Peas, lentil, and beans

S. America

Africa India


Research Question


BibliographyBallinger, Anne. "Causes of Iron Deficiency." Causes of Iron Deficiency. 2006. Web. 25 June 2015.

Bouis, Howarth, and Ross Welch. Biofortification--A Sustainable Agricultural Strategy for Reducing Micronutrient Malnutrition in the Global South. Crop Science Society of America; 2010; 57-66. Ho

Fitzpatrick, Teresa, Gilles Basset, Patrick Borel, Fernando Carrari, and Dean DellaPenna. Vitamin Deficiencies in Humans: Can Plant Science Help?.The Plant Cell; 2012; 92-103.

Gibson, Glenn, and Marcel Roberfroid. "Dietary Modulation of the Human Colonic Microbiota: Introducing the Concepts of Prebiotics." The Journal of Nutrition. Print.

Gilligan, Daniel. Biofortification , Agricultural Technology Adoption, and Nutrition Policy: Some Lessons and Emerging Challenges. Oxford Journals; 2012; 136-148.

Graham, Robin, Ross Welch, David Saunders, Ivan Oritz-Monasterio, and Howarth Bouis. Nutritious Subsistence Food Systems. Advances in Agronomy; 2012; 1-74.

Kirschner, Chanie. What are Prebiotics and do we Need Them? Mother Nature Network; 2013; 1-3.

Micronutrient Deficiency: A Global Challenge to Health. The Journal of Nutrition; 2010; 182-190.

Miller, Dennis, and Ross Welch. Food System Strategies for Preventing Micronutrient Malnutrition. Food Policy. 2013; 17-25.

Pandya-Lorch, Rajul, and Mark Rosegrant. Global Food Security. Nutrition Journal; 2009; 7-24.

Saltzman, Amy, Ekin Birol, Howarth Bouis, Erick Boy, Fabiana De Moura, Yassir Islam, and Wolfgang Pfeiffer. Biofortification: Progress Toward a More Nourishing Future. Global Food Security; 20xx; 9-17.

Sands, David, Cindy Morris, Edward Dratz, and Alice Pilgeram. Elevating Optimal Human Nutrition to a Central Goal of Plant Breeding and Production of Plant-based Foods. Elsevier; 2009; 54-62.

Shergill-Bonner, Rita. Micronutrients. Paediatrics and Child Health. 2013; 77-89.

Tako, Elad, Owen A hoekenga, Leon V Kochian, and Raymond P Glahn. High bioavailability iron maize (Zea mays L.) developed through molecular breeding provides more absorbable iron in vitro (Caco-2 model) and in vivo (Gallus gallus). Nutrition Journal; 2013; 1-11.

Tako, Elad, Raymond P Glahn, Marija Knez, and James CR Stangoulis. The effect of wheat prebiotics on the gut bacterial population and iron status of iron deficient broiler chickens. Nutrition journal; 2014; 1-10.

Tako, Elad, Steve E Beebe, Spencer Reed, Jonathan J Hart, and Raymond P Glahn. Polyphenolic compounds appear to limit the nutritional benefit of biofortified higher iron black bean (Phaseolus vulgaris L.). Nutrition Journal; 2014; 13-28.

Tanumihardjo, Sherry, Howarth Bouis, Christine Hotz, J.V. Meenakshi, and Bonnie McClafferty. Biofortification of Staple Crops: An Emerging Strategy to Combat Hidden Hunger. Comprehensive Reviews in Food Science and Food Safety. 2008; 102-111.

Underwood, Barbara. Overcoming Micronutrient Deficiencies in Developing Countries: Is there a Role for Agriculture?. Food and Nutrition Bulletin. 2000; 44-53.

the effect of plant-based prebiotics on iron

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