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Rapeseed master thesis
Transcript of Rapeseed master thesis
Each organ of the tract is seperated from eachother by muscular sphincters that prevent mixing of material
Food is digested in two ways:
- large pieces are broken into smaller pieces.
- enzymes disrupt bonds between monomers
The Mammalian Digestive System
Most fungi are
(they eat dead things).
Some fungi are
Exercise caution when thinking about eating fungi.
With the exception of Carbon Dioxide and Oxygen, plants recieve most nutrients from the
techniques are utilized to determine the effects of specific nutrient deficiencies on plant growth.
While all organisms require SPONCH to live, plants are particularly sensitive to Potassium deficiencies as well,
Soil is a complex mixture of organic and inorganic compounds of both biological and physical origin.
Soil is produced and replenished through natural processes including deposition, weathering and decomposition.
There is a directionality to typical soil production, with newer, nutrient-rich soil being deposited in the top
The "Dustbowl" in the American Midwest: An example of accelerated
due to human ignorance combined with natural drought cycles.
: one example of a practice that can slow erosion due to farming.
Plant roots are the structures responsible for soil nutrient absorption.
: Major absorptive surface of roots.
Protons are pumped out of plant roots into the soil.
The protons displace nutritive cations (K+, Mg++, etc.) in the soil, which are then absorbed into the roots.
Nutrients move in to roots via diffusion, mostly.
Plants rely upon bacteria and fungi to provide them with some nutrients in the soil.
Bacteria play a major role in the
The action of
bacteria in the soil converts atmospheric nitrogen into biologically useful forms, which can be incorporated from soil into amino acids and nucleotides by plants.
This is how all nitrogen enters the food chain.
: plants that have specialized
adaptations that contain populations of nitrogen-fixing bacteria.
: Symbiotic fungal-root associations.
Increase nutrient absorption by roots.
Development & Structure of Root Nodules
Live in nutrient-poor soils.
Adapted to catch and digest small animals as a supplementary source of soil nutrients (particularly nitrogen).
Plants that live on other plants.
Typical in rainforests and other soil-poor areas.
Can be mutualistic, commensal, or parasitic.
There are many different plant parasites of other plants, with a wide diversity of parasitic adaptations.
Animal nutrition involves four phases:
: Intake of food
: Breakdown of food macromolecules
: Transfer of nutrients into the body
: voiding of undigested materials
Diploblastic animals (cnidarians) utilize a
The "Bag" plan: The mouth of the animal is also the anus.
Triploblastic animals (everything else) utilize a
The "tube" plan: The mouth is at one end of the tract and the anus is at the other end.
The evolution of a gastrointestinal tract allows for compartmentalization of the digestive system and increased efficiency & control of nutrition.
: involuntary contractions of smooth muscle that line the gastrointestinal tract and move food through the tube.
There are a wide variety of strategies used by animals to accomplish this process
Animals need to aquire all nutrients from their environment.
This includes macromolecules, vitamins and minerals.
Some can be synthesized from raw materials, others must be consumed "pre-made"
: Organic, non-macromolecular, compounds
: inorganic elements
Data from an experiment (right) looking at the effects of invasive garlic mustard (left) on the ability of native plants to form mycorrhizal associations
Deficiencies in nutritional requirements will have effects on the physiology of the organism
results in digestion of the bodies stored nutrients.
Lack of specific nutrients will have specific effects on physiology. Shown:
, a protein deficiency
Data showing a correlation between consumption of folic acid vitamin supplements and a decreased likelihood of neural tube birth defects in a sample of the female population of Great Britain.
The human digestive system is a very typical example of the mammalian digestive system.
Teeth, tongue, and salivary glands.
: mechanically digest food.
: moves food into gastrointestinal tract.
which moistens food and contains
(chemically digests starches)
A tube that connects the mouth to the stomach.
Begins transport of food through the GI tract.
: beginning of esophagus.
: Prevents movement of food into the respiratory system
A muscular pouch.
Capable of rapid expansion.
Filled with "
" (a mixture of pepsin and hydrochloric acid).
Processes food into
- Sanitizes food.
- chemicall digests protein
The walls of the stomach contain many "gastric pits"
Three cell types:
: make a protective mucous coating for the stomach epithelium.
: Produce HCl
: Produce the pepsin protein precursor pepsinogen
is an example of a "
", an inactive protein molecule that is converted into the active form in specific conditions
In the case of pepsinogen, HCl is needed to cleave it in to active pepsin.
Why is the pepsinogen/pepsin system necessary?
Term refers to any organs that make secretions that are introduced into the GI tract.
fat (physical digestion).
Excess bile is stored in the
, which contains a variety of hydrolytic enzymes for all macromolecules
The pancreas also produces
, which raise the pH of the chyme (pancreatic enzymes function in basic pH)
Bile and Pancreatic fluid enter the GI tract through a common duct at the beginning of the small intestine (the "
: Digestive section.
: Primary absorptive section (most nutrients)
: Final absorptive section (bile salts and some vitamins).
Small Intestine Ultrastructure:
The epithelium of the small intestine is adapted for absorption of nutrients.
: projections of epithelial tissue. Each vilus is covered in
A network of
(circulatory system) and
(lymph vessels) runs through each vilus, separated from the
(interior) of the small intestine by the brush border.
Nutrients diffuse through the brush border, into the circulatory/lymphatic system.
Carbohydrates, amino acids, and other water-soluble molecules are absorbed into the circulatory system, while lipids are absorbed in to the lymphatic system.
Blood and lymph flow from the small intestine to the liver for detoxification of absorbed molecules.
Final stages of digestion and main area of absorption of nutrients
A tube that holds undigested, unabsorbed digestive material.
Also holds a massive colony (~90 trillion cells) of symbiotic bacteria.
Reabsorption of water.
Production & absorption of vitamins by bacteria.
Storage and (usually) voluntary elimination of undigested food ("feces")
There are several hormones involved in the digestive system.
The GI tract is under the control of an entire division of the autonomic nervous system (the "
" division), and is subject to several different regulatory feedback loops
2. Regulation of Blood-Glucose Level
Controlled by 2 pancreatic hormones:
- removes excess glucose from the blood via storage in body cells and conversion to
in the liver.
- increases glucose level in the blood via glycogen breakdown and glucose release from the liver.
1. Digestion Control
1. The expansion of the stomach to accomodate incoming food trigers the release of
, which stimulates production of gastric juice.
2. As chyme moves in to the duodenum, the presence of amino acids and fatty acids trigers the release of
by duodenal cells, which causes the liver and pancreas to release secretions into the GI tract. The hormone
is also released by the duodenum which causes the pancreas to release bicarbonate ions to neutralize the acidic chyme.
3. Very fatty foods will cause a large amound of secretin and CCK to be released, which has an inhibitory effect on peristalsis and slows down the digestive process.
is the feeling of being "full".
Under the control of the hypothalamus in the brain.
are three hormones that decrease the hunger sensation.
increases the hunger sensation and decreases saitety.
These hormones work in an antagonistic fashion, similarly to insulin and glucagon.
Data from an experiment in which ob mutant mice (genetically prone to obesity) were surgically joined to non-mutants.
ob mutants show decreased body mass gains when joined to non-mutant mice.
While humans have a fairly typical digestive system, other mammals demonstrate various adaptations.
Variation in tooth structure allows animals to adapt to particular sources of food.
GI Tract Adaptations
Many herbivores have highly adapted, elongated appendix structures ("
"), which serve as locations for colonies of bacteria that can aid in cellulose digestion.
Carnivores do not have these structures.
Herbivores that possess highly adapted, expanded upper GI tracts.
Allow for maximized
of vegetable matter and prolonged exposure to symbiotic bacteria.
Temporary obesity is frequently seen in animals to deal with environmental fluctuations and nutritional requirements of specific life stages.
Disruptions in food cycles that animals have adapted to can lead to unintended obesity.
This is perhaps the major reason why there is currently an obesity "epidemic" in the United States.
There are many disorders of the digestive system. Here are a few examples:
"Upper GI" Disorders:
"Lower GI" Disorders:
Irritable Bowel Disease
A disruption of the protective mucus lining of the stomach.
Often caused by the Helicobacter pylori bacterium. Frequently stress-linked.
Persistent "Burning sensation" in upper abdomen.
Antibiotics, diet modification, possible surgery.
Movement of stomach contents through the lower esophageal sphincter.
Transient "Burning sensation" in upper abdomen, particularly following meals.
Antacids, medications that reduce HCl production.
Diarrhea: too much water in feces (decreased absorption by large intestine).
Constipation: too little water in feces (feces remains in large intestine longer than normal).
Immediate: laxatives, indigestion aids (e.g. pepto bismol).
Long-term: Diet modification. Increased fiber intake. Decreased intake of fatty, sugary foods.
Irritation and inflammation of the large intestine.
Ultimate etiology is unclear (stress? Autoimmune?)
Irregular, spastic bowel movements. Discomfort, bleeding (severe cases)
Medication, Diet modification, surgery (extreme cases).
Make Sure You Can:
Why do organisms need to acquire nutrients?
How do organisms acquire nutrients?
What happens if organisms are unable to acquire nutrients?
The Amanita muscaria mushroom causes liver failure and psychosis in animals
"Nom Nom Nom"
"Nom Nom Nom"
Chemical Digestion: A summary
Explain why organisms require nutrients, how they accomplish the processes involved in nutrition, and the consequences of malnutrition.
Compare and contrast plant and animal nutrition.
Explain the structure and function of all organs involved in plant and animal nutrition.
Describe the roles of symbiotic relationships in both plant and animal nutrition.
Describe the modes of hormonal regulation of mammalian nutrition.
Explain the causes, effects and treatments of various disorders of the human digestive system.
"all that remains is a husk"
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Sound effects are a bit over the top
Hamster on a piano...eating popcorn
From bad to worse...
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Rapeseed meal innovative aspects
Aims of the study
- Materials and methods
- Results & Discussion
- Conclusion & References
Internship under the
program held at the
University of Hohenheim - Institute of Animal Nutrition
. Stuttgart, Germany.
Rita Cabrita, Assistant professor at FCUP
Rainer Mosenthin, Full Professor
Institute of Animal Nutrition
(University of Hohenheim)
10^10 – 10^12 cells per gram of feces
Interactions between host <-> GI microbes
Nutrient processing / fermentation
Human gut microbiota
GI = gastrointestinal
Commensal / beneficial bacteria
Pathogenic / pathobiont bacteria
Gut microbiota balance
Human gut microbiota
Principal and adaptable polysaccharide fermenters: genus
Human gut microbiota
YOUR DIET IS LIKE A BANK ACCOUNT.
Important gut microbiota species
Predominant bacterial groups:
Antibiotic resistance mechanisms (beta-lactam antibiotics)
Influence the host immune system to control other (competing) pathogens
Generally beneficial relationship with the host.
Acid production as metabolic end products (acetate, lactate) -> lowers pH -> antibacterial effect
Metabolic end product -> directly inhibitory to a range of Gram(+) and Gram(-) pathogenic bacteria
Complex and diverse group
Synergize with other intestinal microbiota -> ferment unabsorbed dietary carbohydrate -> SCFA (butyrate -> major energy source) -> intestinal epithelial function
ery harmful and responsible
Improves lactose digestion in fermented foods into a more suitable form
Stimulation of immune functions
Lactic acid production -> lowers the gut pH -> reduce pathogenic load
Among the most Gram(-) pathogens associated with respiratory and urinary tract infections
Though, some strains have also positive effects, depending on its amount and the host health (e.g.
“A prebiotic is a
food ingredient that
affects the host by selectively
of one or a limited number of
in the colon, and thus
Non-starch polysaccharides (hemicellulose, pectins, gums and non-digestible OLS)
Carlsson et al. (1992)
High-temperature capillary gas chromatography method
Quantify OLS in foods, diets and intestinal contents
Rapeseed (RS) contains considerable amounts of OLS raffinose family, specially stachyose (0.94-1.52%)
Bifidobacterium growth stimulation
RS is the most important
2nd most important worldwide after soybeans (
RS is currently planted on
of the agricultural cropland
RS cultivation increased 10% Northwest Germany -> increasing
Included in compound feed as dietary ingredient to animal feeding
used for plant oil fuel and bio-diesel
nondigestible dietary carbohydrates
lactose, raffinose, stachyose, fructooligosaccharides (oligofructose; inulin)
By-product of RS oil extraction
Contains up to
protein (dry basis)
Richness in phenolic and other bioactive compounds:
tocopherols, vitamin B, choline
Ca, Mg, Zn, Cu
-> may impose
positive health effects
RSM anti-nutritional factors
reduce feed acceptance
due to their ‘
taste (similar to that in mustard)
Glucosinolates - GSL
Erucic acid - EA
Phytic acid, tannins, mucilage
Plant breeders -> reduce the unwanted ingredients -> “00-RS"
(for farm animals)
Organic sulfur compounds
in farm animals:
Poisoning / toxic
RS for human nutrition
Fleddermann et al. (2012)
Protein-rich residue from RS oil (normally used as animal feed) -> nutritionally equivalent to the protein isolate from soy.
(cultivated in USA) -> may be potentially replaced by
protein harvested in
Effects on the
for human nutrition?
Considering RSM composition and OLS content
Aim of the study
Effects of differently heat treated RSM in the microbial communities in ileal digesta and fecal samples of pigs (
Material and Methods
1. Animals, housing and surgical procedure
Eight (6 + 2 for replacement)
(German Landrace × Piétrain)
10 - 15 days adaptation
period -> pigs were surgically fitted with a simple
Recovery period: 7 days
Material and Methods
2. Diets, feeding and sampling
-> fed level splitted in 2 meals (each time
Each experimental period ->
6 days adaptation
to the diets and fed allowances
(as-fed) of the average BW -> determined on day
of each experimental period
-> fresh feces of each piglet -> sampled immediately
-> ileal digesta sampled at the time of
highest digesta flow rate
Ileal digesta (
) and feces (
) samples into plastic tubing -> stored at
until further analysis.
Material and Methods
3. Bacterial DNA extraction
2 protocols combination:
QIAamp® DNA Stool for Pathogen Detection
Yu and Morrison modified protocol
DNA quantity and quality:
(2x + mean values calculated)
DNA fragments checking:
Agarose gel electrophoresis
1% TAE buffer + 1μul
of 1:100 solution of
Ribosomal Database Project
iCycler iQ5 Real-time Detection System
iCycler Optical System Interface software (Version 2.0)
95°C for 15 min
95°C for 15 sec
primer annealing for
72°C for 20 sec
72°C for 5 min
(final elongation step)
(melting curve data)
1% TBE buffer + 1μul
of 1:100 solution of
5. DNA purification
6. Statistical analysis
SAS STAT software (Version 9.2)
treatment, periods and animals
period × animals effects
(compound symmetry variance-covariance structure)
log10 16S rDNA
gene copy numbers/gram of fresh
Ileal digesta microbiota was very
(gene copy numbers)
for all the bacteria groups studied caused by the different rapeseed treatments, periods or animals either
1. Microbiota activity
due to specific
in rapeseed (e.g. OLS, pectins, arabinose-based carbohydrates)
Different bacterial groups might have been stimulated
2. Pigs' gut microbiota
Index of a health promoting microbiota
3. Heat treatment
4. Prebiotic concept
Results not in accordance with Dinotos’ work.
Probably due to different methods used and sample origins (rat vs. pig)
Host health ->
increase of Enterobacteria
for both period and treatment effects
structure of some nutrients could have been
available and some
Temperature affected the GSL content
-> influenced the nutrients that reached the colon and were fermented by the microbiota
group -> higher growth with
grew more in 6umol GSL/g DM (
Besides some unexpected results:
is a potential candidate as a
raffinose, stachyose, pectins
-> could reach the intestine for
fermentation by intestinal bacteria
Potentially helping to improve the humans and animals gut health
General -> microbiota was very
strongly influenced by the different RS treatments.
showed statistical relevance to the
appeared to have an influence in the
Further analysis ->
optimal RSM processing conditions
; determination of the
initial host microbiota
of the RSM in the
and its use as a
GoLive Probiotics & Prebiotics
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