metabolism Flashcards
proteins digested into
amino acids
polysaccharides digested into
monosaccharides
TAG digested into
fatty acids
what organ plays a central role in processing and distribution of nutrients, and supplies nutrients to tissues via bloodstream
liver
Autotrophs
(“self-feeders”) make organic materials from inorganic materials in the environment
* The biosphere’s producers — Plants
Heterotrophs
(“other-feeders”) use compounds produced by others
* The biosphere’s consumers — Animals
What is Metabolism
Metabolism is the sum of the chemical reactions that convert nutrients
into energy and complex molecules within cells
Metabolism comprises hundreds of chemical reactions, catalysed by _____, organized into discrete ________ which operate in an integrated and coordinated manner
enzymes
metabolic pathways
flows of metabolic pathways
inputs; outputs; passage along pathways
pools of metabolic pathways
amounts of molecules
Filled (supplied) or emptied (by demand) by catabolic and anabolic pathways
space and time of metabolic pathways
not all pathways necessarily occur at the same time, or in the same place
3 main energy-containing nutrients
carbs, fats and proteins
3 reasons metabolism must be regulated
- to prevent “futile cycles”
- to respond to physiological needs
- to respond to changes in energy demand
all metabolic pathways must have overall ____ free energy
negative
can anabolic pathways simplu reverse catabolic pathway
NO
glycolysis, substrate and product
1 glucose –> 2 molecules of pyruvate, makes 2ATP
glycolysis takes place in
cytosol
pyruvate can yield (3 things)
- Ethanol — by anaerobic fermentation in yeast
- Lactate — by reduction in anaerobic conditions in muscle
- Acetyl CoA — by oxidation in aerobic conditions
acetyl coA is a 2C acetyl croup esterified to
co-enzyme A
aceytl coA has a central role in
metabolism
acetyl coA is put into CAC and what energy do you get out
NADH
what is beta oxidiation
breakdown of fat to get NADH; fatty acid trimmed 2 C at a time to get acetyl coA
where does CAC occur
mitochondria
where does beta oxidation occur
mitochondria
3 main VFA
acetate
propionate
butyrate
enzymes from _____ and _____ digest dietary peptides and amino acids (protein)
stomach and pancreas
Protein breakdown (hydrolysis back to amino acids) occurs constantly in cells by two main pathways:
- Cytosolic pathway: involves ubiquitin and the proteasome
- Lysosomal pathway: proteins are taken up by lysosomes and hydrolysed by
proteases (cathepsins)
amino acids split into
amine group –> urea cycle –> urea
and
carbon skeleton –> multiple fates such as ketone bodies, acetyl coA, glucose, CO2
what is purpose of CAC etc making NADH
to produce lots of ATP; in oxidative phosphorylation (ETC)
NADH used to pump protons in to out of mitochondria; creates high proton concentration out of cell, then ATP synthase brings protons back in and this energy phosphorylates ADP into ATP
is pyruvate dehydrogenase reversible
NO its irreversible
can acetyl coA turn back into glucose
NO
gluconeogenesis
Plants: 3-phosphoglycerate
(Calvin cycle) → glucose
Animals: non-carbohydrate
precursors → glucose
– important in fasting
fats breakdown vs fatty acid synthesis
breakdown (beta oxidation)
location: mitochondria
coenzyme: NAD+/ FAD
enzymes: 4
synthesis
location: cytoplasm
coenzyme: NADPH
enzymes: 2
protein synthesis
Ribosomes translate mRNA and synthesize protein
enzymes lower
activation energy
mammalian genome has ~____ genes that code for proteins
30 000
in eukaryotes can one gene code for several proteins
yes
* post-translational modification
* alternate splicing of different exons in the mRNA transcript
To control the amount of a protein in a cell
– control gene expression
(stable vs inducible genes: transcription factors controlled by metabolites, hormones, etc) and
- control protein degradation
Some proteins live long, some ~few minutes
Covalent modification alters protein _____ → regulates activity
conformation
what is a reversible way of regulated proteins
Common important modification: phosphorylation (addition of phosphate group) of serine, threonine or tyrosine side chains
protein kinases
phosphorylates proteins; adds a phosphate group, which changes the structure of proteins therefore changing the activity of that
protein
Each has a specific target protein (substrate) that it phosphorylates, & specific effectors; some have broad, some
narrow, specificity
what is an irreversible way to regulate proteins
activation of a precursor: ie inactive forms of proteins exist and then can be activated (irreversible)
protein kinase examples and what they are activated by
- protein kinase A; activated by cAMP
- phosphorylase kinase; activated by protein kinase A and Ca2+
- protein kinase C; activated by Ca2+
- protein tyrosine kinases; activated by insulin receptorsand others
protein phosphatases
enzyme that cuts off phosphate groups from proteins (opposite of protein kinases)
protein control of activity by non-covalent binding of
effector; may enhance or inhibit activity (allosteric regulation like a built in regulatory network, feedback)
- Allosteric activation, inhibition
- Competitive inhibition
timescale:
less than 1 second
- Activation of preformed precursor proteins
- Activation/inactivation by reversible covalent
modification
time scale
seconds to minutes
- Synthesis of new protein in response to signals (e.g. hormones)
timescale
minutes to hours
- Major changes to the overall protein profile of a tissue (e.g. in response to dietary changes, exercise); rebalancing of synthesis and breakdown
time scale
days to weeks
Erythrocytes
carry O2 from lungs to tissues and CO2 from tissues to lungs
Plasma
carries nutrients (glucose, amino acids, nucleosides, etc) around the body for uptake by tissues; and
Carries metabolites/waste products to (e.g. toxins, glutamine) and from (e.g.
urea) liver; urea goes to kidney for excretion. Also carries hormones
blood delivers what nutrients to brain
glucose, ketone bodies
blood delivers what nutrients to cardiac muscle
glucose, FA, ketone bodies
blood delivers what nutrients to skeletal muscle
glucose, FA, ketone bodies, amino acids
plasma proteins are involved in
blood coagulation and fibrinolysis
albumin (plasma protein)
carries fatty acids and many other molecules
lipoprotein (plasma protein)
carry TAGs and cholesterol esters
Most plasma proteins are synthesized in the
liver
After absorption from the gut, sugars (monogastrics; mainly glucose) and amino acids, VFAs, and some TAG, pass via the blood to the
liver
most TAG is stored in
adipose tissue via lymphatic system
(some in liver)
Hepatocytes transform nutrients into
fuels and precursors for other tissues
Kinds & amounts of nutrients supplied by the
liver vary with
diet and the time between feeds
liver: Demand by non-hepatic tissues depends on
the organ and on the activity of the animal
why does the liver have remarkable metabolic flexibility
- Builds up stores when fuel is plentiful; releases when needed
- Interacts with other organs via the blood, helped by hormones
GLUT2
glucose transporter ensures that hepatic glucose
concentration is the same as in blood
Glucose is phosphorylated by _____ to glucose-6-phosphate when it gets into liver
glucokinase (glucose-6-phosphate is negative and large and basically traps the glucose inside the liver cell)
then turned to glycogen
liver stores glucose as
glycogen
adipose tissue stores ____ and supplies ____
TAGS
fatty acids
adipose tissue is ___% of young mammals weight
15-25%
do all animals need blood glucose
YES; RBCs and brain rely on it!
cats and ruminants blood glucose
blood glucose doesn’t come from the gut
cats: * No correlation with food ingested in the previous 2h * Both normal cats and diabetic cats
ruminants: * ~80–90% of absorbed VFAs are taken up by the liver * Major consequences for hormonal control
rumen microbes ferment
cellulose –> VFAs
horse digestion
digestible carbs –> stomach –> glucose
fat –> small instestine –> FAs
fermentable fiber –> large intestine –> VFAs
rumen microbes
Cellulytic bacteria, protozoa hydrolyze cellulose
rumen microbes turn cellulose
–> cellobiose –> glucose –> VFAs
fate of VFAs
bloodstream –> oxidized –> energy
also –> amino acids and vitamins
acetate
VFA, for energy and FA synthesis
propionate
VFA, forms glucose in gluconeogensis
butyrate
VFA, for energy and FA synthesis, some metabolized in rumen wall and liver then to tissues
VFA absorption
Passive diffusion
* 75% reticulo-rumen
* 20% omasum and abomasum
* 5% small intestine
bloat
lush pasture –> increase sugar –> increase gas
cats blood glucose comes from
glucogenic amino acids (amino acids from diet that have gone through gluconeogenesis)
horses and ruminants blood glucose come from
VFAs and glucogenic amino acids (amino acids that have undergone gluconeogenesis)
gluconeogenesis is active during
fasting
3 main substrates of gluconeogenesis
- Amino acids (from muscle protein breakdown)
- Glycerol (from fat breakdown)
- Lactate (from anaerobic glycolysis).
can fatty acids produce glucose
no!
what is the primary energy substrate
carbs –>
monogastrics: glucose,
ruminants; VFAs
what is primary substrate for fat synthesis
carbs –>
monogastrics; glucose
ruminants; acetate
extent of glucose absorption from gut
monogastrics; extensive
ruminants; little to none
cellular demand for glucose
non-ruminants= high
ruminants and cats= high
importance of gluconeogenesis
monogastrics= less importants
ruminants and cats= very important
intense exercise effects on cell
- cells need to generate lots of ATP
- means its taking glucose-6-phosphate and taking in through glycolysis to generate ATP
- J large
resting state (idling) effect on cells
- not much glycolysis is happening
- also not much gluconeogenesis happening
- these relatively balanced
- J is small and balanced
J= vf=vr
vf= glycolysis
vr= gluconeogenesis
is we had both fructose 6-phosphate (the enzyme for glycolysis)
and fructose 1,6 biphosphate (enzyme for gluconeogenesis) both active at same time in cell what would we get
futile cycle (constant unnecessary back and forth) making and burning ATP, no gain
how does substrate cycling (ie futile cycles) allow fine control of metabolism
- Substrate cycling with no flux through glycolysis uses up ATP for no apparent result BUT:
- Heat is generated
- The ADP produced needs to be reconverted to ATP
in mitochondria ____ and ____ are tightly coupled
oxidation and phosphorylation
in mitochondria, NADH and FADH2 cannot be oxidized unless ____ is present
ADP
when would uncoupling of oxidation from phosphorylation occur
Uncoupling occurs in the brown adipose tissue of animals that live in very cold climates (non-shivering thermogenesis)
neonates have lots of brown adipose tissue
describe uncoupling of oxidation from phosphorylation in mitochondria
protons being pumped out like usual but instead of flowing back through through ATP synthase and phosphorylating ADP –> ATP, instead a protein is there that allows protons to flow freely through different pore
this generates heat!!
highest to lowest capacity for ATP production
OPPOSITE OF fastest to slowest ways to make ATP
- Aerobic lipid metabolism (slowest but most ATP)
Fatty Acid –> Acetate –> CO2 + H2O - Aerobic carbohydrate metabolism
Glucose –> Pyruvate –>CO2 + H2O - Anaerobic glycolysis
Glucose –> Pyruvate –> Lactate - Substrate-level phosphorylation (fastest but least ATP)
Phosphocreatine + ADP –> Creatine + ATP
fastest to slowest ways to make ATP
OPPOSITE OF highest to lowest capacity for ATP production
- Substrate-level phosphorylation (fastest but least ATP)
Phosphocreatine + ADP –> Creatine + ATP - Anaerobic glycolysis
Glucose –> Pyruvate –> Lactate - Aerobic carbohydrate metabolism
Glucose –> Pyruvate –> CO2 + H2O - Aerobic lipid metabolism (slowest but most ATP)
Fatty Acid –> Acetate –> CO2 + H2O
at rest muscles use ___% of O2
working they use up to ___%
50
90
contracting muscles
ATP splits, → energy → fibre contracts
Transfers high-energy Pi → contracting element
ATP → ADP + Pi
** need to be able to regenerate ATP
3 sources of ATP for muscle contraction
- Phosphocreatine (PC)
- Glycolysis
- Oxidative phosphorylation
Phosphocreatine (PC) positives and negatives as a source for ATP for muscle contraction
positives
* Very quick: 4–5s > aerobic
* One step → energy
Negatives
* Little PC stored, used up quickly
rest to exercise oxygen transition
*O2 uptake ↑↑ → steady state ~1–4 mins
* O2 deficit as work begins
* Lag in O2 uptake
∴ Anaerobic glycolysis → ATP
* Steady state: aerobic metabolism ® ATP
very quick exercise (couple of seconds)
phosphocreatine as source of ATP
short burst of exercise (couple of mins) source of ATP
anaerobic metabolism (ie glycolysis)
long exercise (hours) source of ATP
aerobic metabolism
carb sources during exercises
- blood glucose
- muscle glycogen
fat sources during exercise
- plasma FA (from adipose tissue lipolysis)
- intramuscular triglycerides
protein sources during exercise
small contribution to total energy
blood lactate sources during exercise
gluconeogenesis via cori cycle
cori cycle
muscle –> lactate –> liver gluconeogenesis –> glucose –> muscle
metabolic cooperation between liver and skeletal muscle
muscle: glycolysis –> ATP –> contraction –> lactate into blood
liver: ATP used –> glucose –> blood (more expensive)
cori cycle: muscle –> lactate –> liver gluconeogenesis –> glucose –> muscle
ie muscle uses glucose and creates lactate
liver uses lactate and creates glucose
what is primary fuel in low-intensity exercise
fats (needs aerobic conditions ie needs oxygen)
what is primary fuel in hgih-intensity exercise
carbs
crossover concept: Fat → carb as exercise ↑↑, why
- Recruit fast muscle fibers
- ↑ blood epinephrine
in prolonged exercise; CHO
–> fat metabolism
advantages of TAG for storing energy (4)
- Highly reduced: More energy
- Nonpolar: Anhydrous fat droplets: compact
- More space: Larger total energy store
- Insulation
disadvantages of TAG as energy store
- hydrolysis –> FA
- not flexible as energy source ie some tissue can’t use (brain and RBCs)
- cannot form glucose
- not water soluble; inconvenient to move around body
advantages of polysaccharides (glycogen and starch) for energy storage
- most flexible energy source
- very polar, soluble
- hydrolysis –> glucose
- glucose stored as glycogen can be very branched and compact
disadvantages of polysaccharides (glycogen, starch) for energy storage
- bulky since hydrated
- energy content less than TAGS (since its partly oxidized due to have OH groups compared to TAG which have fully reduced FA)
glycogen is a polymer of
glucose
glycogen: a chain of
glucose
can be branched; lots of end points so many places glucose can be added or removed
pretty bulky
glycogen synthesis
glucose phosphorylated; traps it inside cell
added to UTP: activates it into UDP-glucose (activated glucose)
then added to existing chain of glycogen (chain of glucose)
each of these steps has an enzyme that catalyses it
can debranch this chain if there is a demand of glucose (enzyme glycogen phosphorylase does this)
glucose homeostasis during exercise is mediated by hormones like
- noradrenalin (NE), epinephrine (E)
- insulin, glucagon
- thyroxine, cortisol (growth hormone)
hormones
small molecules of intercellular communication
3 modes of action of hormones
- autocrine; act upon themselves
- paracrine; acts locally
- endocrine; everywhere else, ie acts far away via bloodstream
hormone classes based on chemical structure
- steroid hormones
- peptides
- amino acid derivatives
hormones alter metabolism in ____ cells
target
in presence of high blood glucose (lots of glucose in blood) _____ is released by pancreas
insulin
what does insulin do
when there is high blood glucose, insulin counteracts this by lowering the concentration of glucose in blood
does that by telling organs to take up glucose (stimulates liver, skeletal muscle and adipose tissue to take up glucose into their cells)
what does liver do when stimulates by insulin
takes up glucose from the blood into its cells and stores it as glycogen
- increases glucokinase; enzyme that does glucose uptake
- increases glycogen synthase; enzyme that does glycogen synthesis
- inhibits glycogen phosphorylase; enzyme that does glycogen breakdown
what does skeletal muscle do when stimulated by insulin
takes up glucose from blood, store it as glycogen
- increases glucose transporter; enzyme that does glucose uptake
- increases glycogen synthase; enzyme that does glycogen synthesis
- inhibits glycogen phosphorylase; enzyme that does glycogen breakdown
what does adipose tissue do when stimulated by insulin
takes up glucose from the blood, it is turned into fatty acids and glycerol, and stored as TAG (fat)
when there is low blood glucose, ____ is released from pancreas
glucagon
what does glucagon do when released from pancreas
- released when there is low blood glucose
- purpose is to raise glucose concentration in blood
- does this by stimulates liver, skeletal muscle and adipose tissue to release glucose
what does liver do when stimulates by glycogen
- glycogen is depolymerized back into glucose, which is excreted from liver cells into blood
- activates glycogen phosphorylase; enzyme that stimulates glycogen breakdown
- inhibits glycogen synthase; enzyme that does glycogen synthesis
- inhibits phosphofructokinase-1; enzyme involved in glycolysis
- increases enzymes involved in gluconeogenesis
what does adipose tissue do when stimulates by glycogen
release of fatty acids and glycerol from fat –> back into blood
doesn’t effect glucose concentration directly but then other things such as muscle can use these fats instead of using glucose (alternative energy source)
Blood glucose concentration is tightly controlled To prevent
hyperglycaemia and hypoglycaemia
Blood glucose ____ after meals
increases
Blood glucose ____ as cells take it up and metabolise it
decerases
insulin and glucagon are synthesized in
islets of langerhans; small cell clusters in pancreas
insulin _____ glucose storage
increases (ie stimulates the take up of glucose from the blood and therefore decreases blood glucose)
glucagon _____ blood glucose
increases
type 1 diabetes
auto-immune disease; pancreas does not produce enough insulin
type 2 diabetes
pancreas produce insulin normally but it is ineffective
insulin increases
glucagon increases
glucose storage
blood glucose
glucose is ____ so it needs help to cross cell membrane; this is done by:
polar
glucose transporters: GLUT proteins
GLUT 1
widespread glucose transporter protein
GLUT 2,3,4,5,7
tissue-specific glucose transporter proteins
are both insulin and glucagon always present in circulation
yes, at different concentrations; always some futile cycle happening
GLUT2; where and rate
liver, endocrine pancreas
rate of glucose uptake proportional to glucose concentration
GLUT 3 where
brain, nerves; high glucose demand
GLUT4 where
insulin-sensitive transporter; only in muscle and adipose tissue
insulin ____ liver glycolysis
promotes
insulin is released when blood glucose concentration is high and so it also speeds glycolysis in order for more glucose to be used up during glycolysis in order to decrease blood glucose concentration
activates key enzymes in glycolysis by dephosphorylating the enzymes
what does insulin do to the enzymes in glycolysis to promote glycolysis
dephosphorylates them (removes a phosphate group) which activates them
glucagon ___ liver glycolysis
slows
glucagon is released when blood glucose concentration is low and so it also slows glycolysis in order for less glucose to be used up during glycolysis in order to increase blood glucose concentration
does thus by phosphorylating the key enzymes involved
what does glucagon do to the enzymes involved in glycolysis to slow the process
phosphorylates the key enzymes (addition of a phosphate group) in order to deactivate them
glycogen synthase and glycogen phosphorylase
glycogen synthase: synthesizes glycogen by allowing activated glucose to be added to the glycogen chain
glycogen phosphorylase; takes a glucose off the glycogen chain and adds a phosphate group back on: turns it into glucose-1-phosphate
glucose on terminal groups (end of the glycogen chain on any of branch points) where these enzymes can act
insulin effect on glycogen synthase and glycogen phosphorylase
increase glycogen synthase; ie adds glucose to glycogen and increases glucagon storage
insulin _____ glucagon secretion
suppresses
mechanism of insulin action
- Insulin binds to its receptor, a protein tyrosine kinase, never actually gets inside cell
- change in structure and conformation of receptor inside cell
- changes phosphorylation status of insulin receptor
- downstream events via second messengers
glucagon same shid
insulin release is triggered by
glucose metabolism; ie high blood glucose
insulin _____ glycogen synthase and _____ glycogen phosphorylase
increases
decreases
glycogen stores glucose; insulin when blood glucose is high; activates glycogen synthase to store more glucose to decrease blood glucose
second messengers
small molecules that transmit signals
G protein coupled receptors produces
IP3
normal blood glucose in
monogastrics
ruminants
birds
monogastrics: 5mM
ruminants; 3mM
birds; 14mM
cats and horses blood blucose
- little to no glucose from feed
- rely on gluconeogenesis for blood glucose
in ruminants describe glucagon and insulin after a feed
- BOTH glucagon and insulin increase w feeding; highest 2-4 hours w feed
- BOTH decrease in starvation
- effect on insulin on liver is MARGINAL in these animals
- high glucagon stimulates gluconeogenesis (liver); stimulated from VFAs
IP3 and DAG are both
second messengers
is IP3 polar
yes very, therefore in cytosol
is DAG polar
no, therefore in membranes
second messenger vs intracellular signalling
- 2nd messenger; peptide or amine hormones, like insulin/ glucagon which are proteins; too large to enter cell, attach to receptor outside of cell and rely on second messengers inside cell : alter activity of preexisting enzyme, small impact, fast
- intracellular; steroid or thyroid hormones, can enter cells and hormone-receptor complex acts in nucleus: alter amount of newly synthesized proteins (alter transcription of specific genes), big impact, slow
epinephrine (adrenaline)
- increase heart rate
- blood pressure
- dilation of respiratory passages
- increases glycogen breakdown
- decreases glycogen synthesis
- increase glyconeogenesis
- increase glycolysis (this one doesn’t make sense, everything else similar to glucagon)
- increase glucagon secretion
- decreases insulin secretion
cortisol
- liver: increase gluconeogenesis
- muscle: proteolysis: stimulates release of amino acids
- adipose: stimulates hydrolysis of TAG into fatty acids
- counterbalances insulin
T3 controls
basal metabolic rate (BMR)
T3 binds to
transcription factor RXR/THR –> promoter of some genes
stimulates expression in some genes; alters transcription
mammalian fuel reserves
glycogen (liver and muscle)
TAG (adipose)
protein (muscle)
glycaemic index of food
rate at which glucose concentration in blood increases after eating that food
low glycaemic index diets
- smaller increase in blood glucose after meals
- helps animals lose weight
- increases insulin sensitivity
- increases diabetes control
- feel full longer
- increase physical endurance
high glycaemic index increases
carbs after work
what happens in adipose when low blood glucose
lipolysis:
TAG –> FA+ glycerol
FA can be used in liver, skeletal muscle and cardiac muscle via beta oxidation
during starvation
- no glucose stores, no glycogen
- liver makes glucose via gluconeogenesis from amino acids
- means skeletal muscle is doing proteolysis; protein –> amino acids (this only happens when fat stores run out!!!!! last resort)
- adipose is undergoing lipolysis: TAG –> FA + glycerol
- glycerol is used for gluconeogenesis in liver
- FA can be used as energy in skeletal muscle via beta oxidation
- liver turns FA into ketone bodies via beta oxidation and ketogenesis
- cardiac muscle and brain uses ketone bodies as energy source (FA can be turned to ketone bodies in liver)
ketone bodies are produced in
liver during fasting
in tissues other than liver, ketone bodies can be converted into
acetyl coA and fed into CAC and oxidative phosphorylation
2 main ketone bodies
acetoacetate
beta-hydroxybutyrate
leptin is greek for
thin
during starvation describe what happens in CAC
since malate, one of the substrates of CAC can be used for gluconeogenesis, this will occur and then malate will not turn into oxolacetate and acetyl coA will start to accumulate and turn into ketone bodies
how was leptin found
- in lab mice
- 2 mutant copies of gene (ob/ob) that encodes for a peptide hormone called leptin → eat as if starving even when they are fat
- without leptin; can’t link adipose stores to hunger
- if inject leptin into these mice, they would be normal and lose weight
- db/db mice were obese bc they had an issue with leptin receptor
- inject leptin into these mice and wouldn’t help them
leptin system
- leptin produced in adipose tissue
- leptin receptor in brain
- more leptin in blood; signals brain that fat stores are full
leptin and starvation
- adipose tissue shrinks
- less secretion of leptin
- leptin levels decreased
- lower thyroid hormone (lower BMR)
- lower sex hormone
- increase glucocorticoid; mobilizes energy stores
some obesity relate disease in dogs
- Arthritis
- Hip dysplasia
- Ruptured cruciate
- Congestive heart failure
- Dyspnea
- Dermatitis
- Anal Sac disease
- Hyperlipidaemia
- Hypertension
- Hypothyroidism
- Diabetes
- Cushing’s disease
- Cancer
milk carbohydrate
disaccharide lactose
- one molecule glactose- one molecules glucose
milk fats
mostly TAG globules
milk proteins
caseins, serum or whey proteins
lactation increases metabolism, what hormone is important
growth hormone
ketosis
high levels of ketone bodies in blood
ketosis in dairy cattle
- ketones increased because so much FA in lactating cow
- body thinks its full
- increases acidity in blood
- weight loss, loss appetite, less milk yield
