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I.

THE PERMEABILITY OF CELL MEMBRANES


A. EXPERIMENT PROPER
Hemolysis of Human Erythrocytes
1. Solutions
RBC suspension
o 7 drops blood
o 5 mL 0.9% NaCl
(+) control
o 5 drops RBC suspension
o 5 mL dH2O
(-) control
o 5 drops RBC suspension
o 5 mL 0.2M NaCl
Glucose (5 mL each test tube)
o 0.075, 0.09, 0.1, 0.15, 0.2
M
CaCl2 (5 mL each test tube)
o 0.03, 0.035, 0.05, 0.075,
0.09 M
Na2SO4 (5 mL each test tube)
o 0.03, 0.035, 0.05, 0.075,
0.09 M
KCl (5 mL each test tube)
o 0.025, 0.05, 0.075, 0.1,
0.15 M
NaCl (5 mL each test tube)
o 0.025, 0.05, 0.075, 0.1,
0.15 M
2. Procedure
Add 5 drops of suspension to each
concentration of every solution
Record hemolysis time (complete
hemolysis has occurred when the
solution appears transparent against a
white printed background)
Threshold: 10 minutes
Calculate for Isotonic Coefficient (i) and
Degree of Dissociation (a) see
equations in discussion
Effect of Molecular Size on Permeability
1. Solutions
RBC suspension
o 7 drops blood
o 5 mL 0.9% NaCl
(+) control
o 5 drops RBC suspension
o 5 mL dH2O
(-) control
o 5 drops RBC suspension
o 5 mL 0.2M NaCl

0.3 M urea
0.3 M glucose
0.3 M ethylene glycol
0.3 M glycerol
2. Procedure
Add 2 drops of suspension in each
solution
Record hemolysis time
Threshold: 10 minutes
Permeability and Partition Coefficients
1. Solutions
RBC suspension
o 7 drops blood
o 5 mL 0.9% NaCl
(+) control
o 5 drops RBC suspension
o 5 mL dH2O
(-) control
o 5 drops RBC suspension
o 5 mL 0.2M NaCl
0.6 M methanol
0.6 M ethanol
0.6 M isopropanol
0.6 M n-butanol
2. Procedure
Immerse solutions in an ice bath are
take note of temperature
Add 2 drops of suspension in each
solution
Record hemolysis time
Threshold: 10 minutes
Effect of Polar Groups on Permeability
1. Solutions
RBC suspension
o 7 drops blood
o 5 mL 0.9% NaCl
(+) control
o 5 drops RBC suspension
o 5 mL dH2O
(-) control
o 5 drops RBC suspension
o 5 mL 0.2M NaCl
0.3 M chloroform
0.3 M hexane
0.3 M ethylene glycol
0.3 M isopropanol
2. Procedure
Add 2 drops of suspension in each
solution
Record hemolysis time
Threshold: 10 minutes
ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

B. DISCUSSION
The Cell Membrane
Phospholipid bilayer
Outer: hydrophilic heads, inner: hydrophobic
tails
Other components: membrane proteins,
other lipids (i.e. cholesterol)
Selectively permeable
Regulates the passage of materials into and
out of the cell
o Nutrition
o Maintenance of irritability (i.e.
sensitivity to stimuli, fluidity of
membrane)
o Homeostasis (milieu intrieur)
Transport Through the Membrane
Passive
o Simple diffusion the net movement
of solutes through a selectively
permeable membrane from an area
of higher energy to an area of lower
energy
(higher
to
lower
concentration) by random thermal
motion
o Facilitated diffusion assisted by
proteins embedded in the membrane
but still across a concentration
gradient
Channel proteins provide a
path for lipid insoluble
molecules to pass through
Carrier proteins undergo
conformational changes to
transport solutes
Active
o Occurs against the concentration
gradient and, therefore, requires
energy
o This energy is usually provided in
the form of adenosine triphosphate
(ATP)
Osmosis
o Diffusion of water through a
selectively permeable membrane
o Water moves according to its own
concentration gradient
Osmoticity vs. Tonicity
Osmoticity refers to the relationship of
solutions on opposite sides of an ideal
selectively permeable membrane (one that
2

ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

does not allow the entry of any solutes);


isosmotic solutions have the same number
of particles per liter of solution (osmolarity)
Tonicity refers to the relationship of
solutions separated by a biological
membrane (i.e. plasma membrane); refers
to the response of a cell or tissue; for
example: hypo-, hyper-, or isotonic to the
cytosol
Solutions can be isosmotic but not isotonic;
for example, 0.3M glucose is (theoretically)
isosmotic to intracellular fluid of RBCs but it
still caused hemolysis because of the nature
of the actual biological membrane (allows
solutes through)

Red Blood Cells


Often used model for the membrane-solutesolvent relationship
Hemoglobin pigment in the cell that (aside
from containing oxygen) gives it its
characteristic red color
When hemolysis occurs in a hypotonic
solution because water rushes into the cell,
this pigment spreads through the solution,
giving it a clearer appearance
Isotonic Coefficient

where: CNE is the concentration of the NE that


caused hemolysis within 10 minutes, CE is the
highest concentration of an E that caused
hemolysis within 10 minutes

Characteristic of an electrolyte relative to a


non-electrolyte:
concentration
of
an
electrolyte that exerts an equal amount of
osmotic pressure as a non-electrolyte
Assume the osmotic activity is directly
related to hemolytic point (therefore, HNE
and HE are equal to the NEs and Es
concentrations, respectively)
The value implies that the given
concentration of electrolyte exerts i times as
much osmotic pressure as the given
concentration of the non-electrolyte
A non-electrolytes i is equal to 1, therefore:
o i close to 1 = strong electrolyte
o i < 1, close to 0 = weak electrolyte

Degree of Dissociation

higher partition coefficients diffuse more


quickly through the membrane
where: i is the isotonic coefficient and k is the
number of ions that the E dissociates into in
solution

Solute
Chloroform
Hexane
Ethylene Glycol
Isopropanol

the fraction of the solute molecules that


dissociates in solution
related to the tendency of an electrolyte to
dissociate in solution

Molecular Size and Permeability


Solute
Ethylene Glycol
Urea
Glycerol
Glucose

Polarity and Permeability

Molecular Weight
62.07 g/mol
66.06 g/mol
92.09 g/mol
180.00 g/mol

generally (and logically), bigger molecules


take longer to diffuse into a membrane
there is a direct (positive) relationship
between molecular size and time of
hemolysis

Dielectric Constant
4.81
2.0
37.0
17.9

There is a positive relationship between


polarity and hemolysis time in that more
polar molecules less easily penetrate the
cell membrane
This is due in part to the membranes
composition
(hydrophilic
heads,
hydrophobic tails) and due to its slight
electrical potentialpolar molecules have a
separation of charges that affect the rate of
their uptake by the cell

Partition Coefficients and Permeabilty


Solute
Methanol
Ethanol
Isopropanol
n-Butanol

Partition Coefficient
(k)
0.14
0.26
0.64
7.7

A
partition
coefficient
measures
hydrophobicity in that it is the ratio of a
molecules solubility in a lipid and the same
molecules solubility in water
Overtons
Rule
states
that
higher
hydrophobicity
relates
to
higher
permeability; there is a positive relationship
between the partition coefficient and time of
hemolysis
Non-polar compounds tend to have higher
solubility in water and (way) lower solubility
in water and, therefore, higher partition
coefficients; molecules with longer carbon
chains permeate the membrane more easily
than molecules with shorter carbon chains
Because of the hydrophobic nature of a bulk
of the cell membrane, substances with
ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

II. ACTIVITY OF SALIVARY AMYLASE


A. EXPERIMENT PROPER
Determination of Achromic Point
1. Solutions
Salivary amylase dilution
o Saliva was collected after
stimulation of salivary glands
through the mastication of
paraffin wax
o Saliva was filtered through
cheese cloth then diluted to
10 mL then filtered again
through filter paper
1% Cooked Starch solution
o 5 g starch suspended in 50
mL d H2O then slowly
dissolved with 450 mL boiling
water
o Stirred and heated until
translucent
Iodine-Potassium Iodide solution
(IKI)
o 6 g KI
o 4 g iodine crystals
o 100 mL d H2O
1% NaCl solution
0.2 M Na2HPO4 (for McIlvaines
solutions)
0.1 M citric acid
Phosphate buffer, pH 6.8
Digestive mixture (maintained at
38 )
o 5 mL starch
o 2 mL 1% NaCl
o 2 mL phosphate buffer, pH
6.8
o 1 mL salivary dilution
2. Procedure
1 mL of the salivary dilution was added
to the prepared digestive mixture once it
reached 38 and the time of addition
was recorded
Temperature was kept constant by
placing the test tube in a water bath
At minute intervals, a drop of the dilution
was added to a spot plate with 1 drop of
IKI and color change was observed
If no color change was observed within
2-10 minutes, the number of amylase
units and enzyme activity in the saliva
were computed
Note: 1% starch contains 50 mg starch
4

ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

Effect of Enzyme Concentration


The procedures for determining achromic
point, amylase units, and enzyme activity
were performed for 6 different salivary
dilutions
Enzyme activity vs. Enzyme concentration
was graphed
Effect of Substrate Concentration
The procedures for determining achromic
point, amylase units, and enzyme activity
were performed with the [S] of the 5 mL
starch in the digestive mixture varying from
1% to 6% (use MW of starch 3 x 104 Da to
compute for concentration of solutions)
Enzyme activity vs. Substrate concentration
was graphed
Michaelis-Menten Plot: Reaction velocity
(M/min) vs. Substrate Concentration (M)

Lineweaver-Burke Plot: Reciprocal of


reaction velocity (1/M-min-1) vs. Reciprocal
of Substrate concentration (1/M)

Effect of pH
McIlvaines Solutions
pH
3
4
5
7
8

0.2 M Na2HPO4
(mL)
4.11
7.71
10.30
16.47
19.45

0.1 M citric
acid (mL)
15.89
12.29
9.70
3.53
0.55

Procedures for determination of achromic


point, amylase units, and enzyme activity
were performed with the 2 mL phosphate
buffer, pH 6.2 in the digestive mixture being
varied
Enzyme activity vs. pH was graphed

Effect of Temperature

Procedures for determination of achromic


point, amylase units, and enzyme activity
were performed with the temperature of
digestive
mixture
being
varied
(3
temperatures < 38 < 2 temperatures)
Enzyme activity vs. Temperature was
graphed

B. DISCUSSION
Enzymes

Biological catalysts that enable biochemical


reactions to occur in actual physiological
conditions by lowering their activation
energy

Contain highly specific active sites to which


certain substrates can bind to form an
enzyme-substrate complex

Highly sensitive to pH, temperature, and


reactant concentrations

Regulation by Inhibition:
o Competitive inhibitor binds to active
site
o Non-competitive inhibitor binds to
enzyme at site other than active site
Salivary Amylase
Also known as ptyalin
Hydrolyzes starch and glycogen through its
action on -1,4- glycosidic bonds
Starch dextrins maltose
Optimum pH range: 5.6 to 6.9
Optimum temperature: 38
Starch-Iodine Test
Starch is a polymer of -D-glucose
molecules bound by -linkages; it forms a
helical structure
This structure is significant because it forms
a complex with iodine molecules as the I2
particles nestle themselves within the helix
and for a blue-black complex
In the lab, this can be used to test the
presence of starch in a certain solution
Achromic Point
The point at which no blue-black color is
produced when a drop of digestive mixture
is added to I2-KI
The non-formation of blue, black, or violet is
indicative of the cleavage of the starch in

the digestive mixture into maltose as there


is no helix into which the iodine can insert
itself
Amylase Units and Enzyme Activity
Using the number of minutes it took to reach
achromic point, amylase units and,
consequently, enzyme activity of the
amylase in the collected saliva can be
computed
Amylase units (#AU): the amount of
amylase needed to digest 5 mL of 1%
starch

Note: mL of digestive mixture in our


experiment was 10 mL; dilution factor is
equal to total mL of solution divided by mL
of enzyme (in this case, saliva)
Enzyme Activity (EA): the amount of starch
(in mg) hydrolyzed per minute per unit
enzyme

Enzyme Kinetics
Effect of Enzyme Concentration
o There is a positive relationship
between enzyme concentration and
the rate of reaction or enzyme
activity
o Graph does plateau, however, since
substrate concentration is kept
constant
Effect of Substrate Concentration
o Theoretically, there should also be a
positive
relationship
between
substrate concentration and the rate
of reaction or enzyme activity
o However, the reality is that enzymes
occur in finite quantities in the
bodyeven if the substrate is
abundant,
enzyme-substrate
complexes can only form if there are
available enzymes
Michaelis-Menten Equation

ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

Reaction
Velocity
vs.
Substrate
concentration
The Michaelis-Menten constant (Km)
describes the affinity of the enzyme and
substrate; Km = affinity
At Km, the rate of reaction is at 50% of Vmax

Lineweaver-Burke Equation

1/Reaction
Velocity
vs.
1/Substrate
Concentration
To linearize the M-M plot
Slope of Line Equation = Km / Vmax
x-intercept of line = - 1 / Km
y-intercept of line = 1 / Vmax

Effect of pH on Enzyme Activity


The acidity or alkalinity of the environment
in which the enzyme occurs can affect its
active site (conformational changes may
occur)
Extreme pH can cause the protein to
dismantle (too low and carboxyl ions
protonate; too high and amino groups will
be deprotonated)
Graph is a bell curve as there is an optimum
range
Effect of Temperature on Enzyme Activity
Higher temperatures generally benefit
chemical reactions in that they enable
higher kinetic energy of the molecules, thus
increasing rates of collision and reaction
Graph is skewed to the right as increasing
temperatures can speed up reactions but
after a certain point (i.e. 38 ), the rate of
reaction will decline
Extremely high temperatures denature the
enzymes

ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

III. THE ANIMAL-LIKE Paramecium


A. EXPERIMENT PROPER
Gross Morphology & Movement
Immobilize Paramecium using either 2%
agar or cotton fibers
After noting observable morphology, stain
with methylene blue to visualize macro- and
micronuclei and other features
Digestion
Yeast feed yeast and water were boiled
for 8 minutes then cooled, liquid decanted,
and 4 drops of Congo Red were added
0.1 g dry active yeast
40 mL d H2O
Congo red
o Note: orange red in pH 5.0 or
above, blue in pH 3.0 or below
Slides were prepared using both agar and
cotton fiber method
1 drop of stained yeast feed was added to
cover slip then slide was observed
immediately
Role of Contractile Vacuole
1. Solutions
d H2O
Paramecium culture liquid
2.5%, 3%, 5%, 7.5% 10% NaCl
M/500 KCN (13 mg solute in 100 mL
water)
0.1% neutral red
o Note: red in acidic medium,
yellow in alkaline medium
2. Procedure
Determination of pH of contractile
vacuole contents
o Slides were prepared with cotton
fibers
o 1 drop neutral red onto culture before
laying on cover slip to determine pH
of contractile vacuole contents
Counting the pulsations of contractile
vacuole per minute
o Slides were prepared using both agar
and cotton fiber immobilization
techniques
o 1 drop of the different solutions was
added before covering
o Number of pulsations was counted
under the microscope

Thigmotaxis
Observe Paramecium slide prepared using
cotton fiber method; note difference in
movement when anterior and posterior ends
hit thread
Chemotaxis
1. Solutions
0.2% HCl
0.2% CH3COOH
0.1 M NaHCO3
3% NaCl
5% sucrose
2. Procedure
Prepare Paramecium slides without
cotton fibers and agar
Dip cotton thread into each of the
solutions listed above and lay it on the
slide
Observe reaction of organism
Geotaxis & Phototaxis
Geotaxis
o Test tube was filled of the way up
with Paramecium culture then set
upright in a test tube rack
o Positioned under a bright light for 15
minutes
Phototaxis
o Test tube was filled of the way
with Paramecium culture then
sealed with a rubber stopper
o Tube was laid on its side and its
upper surface was covered with foil
o Light was shone from below the tube
Galvanotaxis
Ringers Solution (1 L)
6.5 g NaCl
0.14 g KCl
0.12 g CaCl2
0.20 g NaHCO3
0.01 g Na2HPO4
4 x 0.5 x 0.3 cm Paraffin well on slide
Fill well with 3-5 drops of Ringers solution
and culture
Attach uninsulated ends of 24-gauge wires
with tape; wires connected to batteries to
close the circuit
Voltage was adjusted to 4.5, 6, 7.5, 9.0, and
12 V using multiple batteries
ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

o
B. DISCUSSION

Paramecium
Kingdom Protista, Phylum Ciliophora;
eukaryotic, unicellular
animal-like = locomotory, no cell wall,
heterotrophic
easily excitable cell membrane

Gross Morphology

o
o

Oral groove gathers food and


leads toward cytoproct
Cytostome or cell mouth opening
of the cell
Cytopharynx or gullet forms food
vacuole for digestions
Transient
food
vacuole:
retains
alkaline
internal
environment
when
not
digesting food
Cytoproct or anal pore used for
digestive waste disposal
Macronucleus performs normal
cell functions
Micronucleus performs specialized
functions like reproduction (asexual
by binary fission and sexual by
conjugation)

Movement
Locomotion
primarily
controlled
by
metasynchronous ciliary beating (effective
stroke propels forward, recovery stroke
keeps cilia flexed)
Spiral motion that appears zig-zag, rotates
(counterclockwise) along longitudinal axis
due to oblique ciliary beating

Slipper-shaped, dorsal surface is convex


while ventral surface is flattened
Anterior end is more blunt, posterior end is
slightly pointed
Cilia line the outside of the Paramecium and
function for movement and gathering and
moving food towards the oral groove
Ectoplasm clear outer zone of cytoplasm
o Pellicle protective layer that
maintains shape
o Cell Membrane
o Myoneme longitudinal muscle-like
striations
o Contractile Vacuoles regulates
amount of water in the cell; expels
water
o Radiating Canals paths that
connect to the contractile vacuoles
(permanent structures)
o Trichocysts defensive structures
that release a fibrous secretion when
under stress or to anchor during
feeding
Endoplasm granulated, inner cytoplasm
zone

ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

Digestion

Oral groove vestibule buccal cavity


overture cytostome cytopharynx
Ingestion: food enters the oral groove then
vestibule then cytostome by movement of
the cilia
Digestion: food is received by the gullet or
cytopharynx, where a food vacuole will form
and bud off of the gullet and starts travelling

around the cell in a clockwise path


(cyclosis); note that it also decreases in size
as it travels around the cell
The food vacuole contains lysozyme that
hydrolyzes its contents, thus shifting the
internal pH to acidic then back to alkaline
(Congo red can be used to visualize this
transition: blue red blue)
Egestion: release of alkaline digestive
wastes through the cytoproct or anal pore
(note that nitrogenous wastes are released
through the cell membrane) and the refusion of the food vacuole with cytopharynx

During diastole, the vacuoles enlarge, while


they collapse in systole

Rate of discharge varies with temperature


Higher rate of discharge occurs when
Paramecium is inactive
Higher rate of discharge occurs when there
is a low salt concentration
By creating a gradient wherein there is
higher salt content in the external
environment, water is able to diffuse out of
the cell
The contents of the contractile vacuole are
acidic and, therefore, should appear red
when stained with neutral red

Effect of Test Solutions on the Contractile


Vacuole
Solution or Medium

Role of Contractile Vacuole


Contractile vacuoles can also be called
water-expulsion vacuoles as they expel
water; they facilitate the excretory pathway
for nitrogenous wastes (urea and ammonia)
They are homeostatic; prevent cytolysis and
osmolysis
Anterior and posterior contractile vacuoles
do not pulsate or beat at the same time;
they are temporary structures
Water passes through the radiating canals
and into the vacuoles before being exuded
through a pore

Distilled water
Paramecium culture
M/500 KCN
2.5% NaCl
3% NaCl
5.0% NaCl
7.5% NaCl
10.0% NaCl

Contractions per
Minute
20
15
6
8
8
7
7
2

As expected, most pulsations occur when


the Paramecium is exposed to distilled
water and least when in 10% NaCl
KCN is poisonous; in this case, it reduced
the production of ATP in the mitochondria
(inhibits NADH oxidase) and affected the
contractions negatively

Thigmotaxis
Negative response to stimuli
ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

Difference
between
reactions
when
opposite ends are touched is due to type of
channels located on each
Anterior end: reverse movement
o Depolarization of membrane by
influx of Ca2+, cilia slow down and
Paramecium reverses
Posterior end: faster forward swimming
o Hyperpolarization of membrane by
efflux of K+, cilia accelerate

Chemotaxis
Defined as the directional response to
chemical stimuli (innate mechanism)
Not to be confused with chemotactic
response, which refers to the reactions of
Paramecium to a new or unknown stimuli
Chemokinesis, on the other hand, is random
change in movement of organism due to
chemical stimuli
Poositive chemotactic response to weak
acids and sugar solutions (acetic acid and
sucrose)
o Paramecium thrive in slightly acidic
environments
as
the
main
component of their diet, bacteria,
regularly occur in slightly acidic
media
o Causes the hyperpolarization of the
membrane
(more
negative
membrane potential) and, thus,
acceleration of ciliary beat
Negative chemotactic response to strong
acids, bases, and salt solutions (HCl,
NaHCO3, and NaCl)
o Salt solutions are hyperosmotic and,
thus, cause the depolarization of the
membrane as water rushes out of
the cell (also, Paramecium is a
freshwater resident)
o Strong acids have adverse effects
on Paramecium; high concentration
of H+ causes depolarization of
membrane
o Bases cause the depolarization of
the membrane due to the low
concentration of H+
Geotaxis
Response to gravity
Paramecium exhibit negative geotaxis
Individual organisms gather near the surface
of the medium
1
0

ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

Mechanoreceptors on their anterior and


posterior ends allow them to reorient
themselves based on the pull of gravity
Buoyancy is also helped by the contractile
vacuole
Phototaxis
Response to light
Positive phototaxis
This response could be an adaptation to
find food (i.e. bacteria)
Light
could
also
follow
increased
temperaturesthe optimal temperature for
Paramecium is said to be around 23
Galvanotaxis
Response to electric current or flow of
charges in a medium
Generally, Paramecium is attracted to the
cathode (negatively-charged pole) since
they are naturally positive outside their
membranes
In weak electrical voltage, the organisms
moved towards the cathode
In strong electrical voltage, the organisms
moved towards the anode then died
Depolarization due to Ca2+ influx causes
ciliary reversal and movement to the
cathode
Hyperpolarization due to the efflux of K+
causes ciliary augmentation or acceleration
and movement to the anode
Ludloff Phenomenon: hyperpolarization of
Paramecium causes ciliary augmentation

IV. OXYGEN CONSUMPTION AND METABOLIC


RATES OF TERRESTRIAL ANIMALS &
MEASURING CARBON DIOXIDE PRODUCTION
IN ANIMALS
A. EXPERIMENT PROPER
1. Balb/C mouse
5-month old male
Weighed using top-loading balance
Inbred albino strain of mice commonly
used in the lab due to their reproductive
efficiency and longevity
2. O2 consumption and CO2 production
measurement
4.53 L-capacity PASCO EcoChamber
o Mouse was acclimatized for 5
minutes
o Lid was sealed with masking
tape and entire chamber was
covered with a lab gown to
minimize mouse stress
PASPORT Temperature Sensor
PASPORT O2 Sensor
o Calibrated to around 20.9%
PASPORT CO2 Sensor
o Calibrated to around 400 ppm
PASCO Scientific DataStudio
o Measured O2 and CO2 content of
metabolic chamber for 10
minutes
3. Values computed for:
Volume of O2
o
converted to L/L

Metabolic Rate
Basal Metabolic Rate (MR in g-cal/hr)

Mass-specific Metabolic Rate (MSMR in


g-cal/hr/g)
=

Volume of CO2

Respiration Rate (RR in M/g/hr)

4. Graphs needed:
O2 concentration vs. CO2 concentration
Mass-specific metabolic rate vs. mass
Respiration rate vs. mass
B. DISCUSSION
Metabolism
Sum of all biochemical activity occurring
within an organism
Related to the amount of energy that the
organism needs to function
Two types:
o Anabolism reductive reaction to
synthesize bigger molecules using
smaller ones; requires energy
o Catabolism oxidative breakdown of
complex molecules into smaller
ones; releases energy
Two pathways based on the presence of O2:
o Aerobic in the presence of oxygen;
36-38 molecules of ATP
o Anaerobic in the absence of
oxygen; 2 molecules of ATP
Measurement of Metabolic Rate
Metabolic reactions transform chemical
energy into heat energy; measure the
amount of heat energy transformed per unit
time measures metabolic rate
Measurement Methods:
o Calorimetry measuring heat output
Direct measuring heat
produced directly (i.e. ice
bath calorimeter)
Indirect based on gas
exchange; assumes that the
intake of O2 and the output of
CO2
corresponds
to
metabolic activity and relates
the concentrations of these
to heat production through
stoichiometry
o Radioisotope Labeling
o Respirometry
Warburg Method

ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

1
1

Pressure
measurement
of
gases
Gilson Respirometer
Absorption of CO2
animal releases

Different Metabolic Rates in Individuals


Standard or Basal Metabolic Rate
o The use of standard or basal to
describe this depends on the mode
of thermoregulation employed by an
animal
o SMR is measured for ectothermic
animals while BMR is measured for
endothermic animals; BMR must be
measured within the endotherms
thermoneutral zone (when they
expend no energy to regulate their
core temperature)
o Measured under 3 conditions:
i. Resting state outside of
reproductive stage
ii. Fasting or in post-absorptive
state
iii. Free from physical, thermal,
and psychological stress (i.e.
not expending any energy to
regulate temperature)
Active Metabolic Rate
o Measured while animal is being
motivated or exposed to a stressor
o Maximum level of activity (i.e.
exercising)
Routine Metabolic Rate
o Measured while animal is unfed and
performs its usual daily activities; in
their natural habit and habitat
Correcting Observed Volume of O2
Based on the effect of actual deviations
from standard pressure and temperature
conditions, 760 mmHg and 0 K,
respectively, on the volume of gas
measured
Note: Ideal Gas Law: PV = nRT
Factors That Affect Metabolic Rate
Mass or Body Size
o A larger animal would require more
energy to maintain its function,
therefore, it would have a higher
metabolic rate
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ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

However, small animals have higher


metabolic and respiration rates per
unit mass so there is an inverse
relationship between mass and
mass-specific metabolic rate
Surface Area to Volume Ratio
o Smaller animals that have a higher
surface area to volume ratio have
higher metabolic rates than larger
animals
o Heat radiates from the animals
skina small animal would gain and
lose more through its skin due to the
lack of solid mass in their body to
retain heatso, in a sense, they
compensate by facilitating higher
metabolic activity
Temperature
o Generally
exhibits
a
positive
relationship with metabolic rate as
oxygen consumption is said to
increase with temperature
o In addition, higher temperatures
favorably affect enzyme activity
during metabolism
Light
o Most drastically affects metabolic
rate through the circadian rhythm of
organisms; diurnal animals are more
active during the day and, thus, have
higher metabolic rates during this
time
o Studies have shown that light may
affect metabolic rate as it stimulates
the release of certain hormones
o In addition, a higher level of activity
usually occurs in well-lit areas (i.e.
fish as they forage)
Sex
o Generally, men have higher rates of
metabolism than women
o Differences are due to male and
female
differences
in
body
composition (fat vs. muscle)

Factors That Affect Respiration Rate


Temperature
o Generally, respiration increases with
higher temperaturehowever, while
this relationship is established, it
does not occur too frequently as
other factors more drastically affect
respiration rate

For ectotherms, their respiration rate


increases while they adjust to their
ambient temperature but, after they
reach physiological temperature, the
rate normalizes or decreases
o For endotherms, temperature is only
a notable factor when they are
inactive
Level of Activity
o Generally, a higher level of activity
results in a higher rate of respiration
(for obvious reasons)
Size
o Generally, larger animals have
higher rates of respiration as their
functioning requires more metabolic
activity

Possible Sources of Error


Calibration error
Stressed mice
CO2 production is not a very accurate way
to measure metabolic rate:
o Aside from exhaled CO2, there is a
large pool of free and dissolved CO2
in the body
o Unlike
O2
consumption,
CO2
production (read: actual metabolic
activity) is substrate dependent

ANIMAL PHYSIOLOGY: FIRST LAB EXAM / A.B.C.

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