Republic of the Philippines

DEPARTMENT OF SCIENCE AND TECHNOLOGY

PHILIPPINE SCIENCE HIGH SCHOOL - CARAGA REGION CAMPUS BIOLOGY 3

LABORATORY ACTIVITY 2
ABIOTIC FACTORS WITHIN THE ENVIRONMENT
Adapted from MSUIIT Laboratory and Field Manual in Bio.107.2 General Ecology
(February 5-7, 2019)

Name:__________________________________ Score:__________________________________
Year and Section:_________________________ Date:___________________________________

Living organisms occupying a given habitat are exposed to conditions which are largely defined by
the nature of the physical environment they inhabit. Thus, various abiotic factors are often prime
determiners of the abundance, distribution and over-all ecology of the biotic units of the ecosystem. The
abiotic components are the complex physical factors which include climatic factors (light, temperature,
humidity, and wind), edaphic factors (soil nutrient, acidity and moisture) and topographic factors (angle of
slope and altitude). According to A. G. Tansley (1935), “the biotioc components could not be separated
from the abiotic with which they form one physical unit, the ecosystem”.

Objectives:
1. To acquaint the students with the different climatic and edaphic factors.
2. To know the standard procedures used in measuring these factors.
3. To determine variations in the physical environment in a small area consisting of grasses and trees.
4. To identify organisms found in both areas.
5. To analyze how individual organisms respond to their physical environment.

Materials:

2 Calibrated string (10 m) Glass rod Concentrated Nitric acid

2 Thermometer Spatula/knife (HNO3)
Meter Stick Droppers Distilled water
Psychrometer Plastic bags/ ziplock 70% ethanol
Aluminum pans Oven Light source
2 Bolo/ Shovel Muffle furnace White paper
Soil corer Crucible or aluminum dish Forceps
pH meter Desiccator Magnifying lens
Digital weighing apparatus Ammonium oxalate Small jar or vial
Gloves 10% HCl Stereomicroscope
Beaker (150 ml) Diphenylamine 2 Millimeter ruler
8 Test tubes Sulfuric acid (H2SO4) Marker
Test tube rack Ammonium molybdate Masking tape

Procedure:

A. Climatic Factors
1. Choose an area with two adjacent types of environment (preferably a grassy area, representing
a sunny area, adjacent to an area with trees, representing a shady area.
2. Lay a calibrated 10m line in each of the two sites. Choose 3 random points within the line.
3. Air Temperature
Measure the air temperature using a laboratory thermometer in each of the sampling points.
Suspend the thermometer 1m above the ground for three minutes before taking any reading.
Record all values in degrees Celsius (oC).
4. Relative Humidity
Use a slingshot psychrometer to get the humidity. Moisten the cloth wrapped around the
end of the wet bulb with clean water. Rotate the psychrometer in the air for 2 minutes, and get
 the wet and dry bulb readings in each sampling point. Use the conversion table (Table 1) to
determine the relative humidity based on the temperature readings.
5. Precipitation
Measure precipitation using 2 calibrated containers (if rain gauge is not available). Leave the
containers in the sampling area for 24 hours. Measure the precipitation (ml/day) based on the
collected water. Alternatively, a precipitation data may be obtained from local weather stations.
6. Evaporation
To measure the rate of evaporation, place an aluminum pan with a known volume of waterin
each sampling point. Record the volume in ml/day.

B. Edaphic Factors
1. Soil Profile
Collect soil samples in the same area using a soil corer. Push the tube into the soil until the
top of it level with the soil surface. Pull it carefully from the soil and examine. Collect enough
information concerning the O, A, and B horizons of the soil. Take note of the differences in
color, structure and thickness within these major horizons. (The corer must be cleaned first
before taking the next sample). Students may also get complete soil profile conveniently from
recent excavations in the area.

Table 1. Percent relative humidity based on dry and wet bulb temperatures ( oC) of a slingshot psychromoter.

2. Soil Temperature
Place the bulb of the thermometer 5 cm and 15 cm from the soil surface to get the soil and
deeper soil temperature, respectively. Repeat in each sampling point.
If the soil temperature profile is to be determined, several thermistors will be buried at different
depths. Temperature profile can be obtained by plotting the soil temperature on the horizontal
axis and depth of soil on the vertical axis.

3. Soil pH
 Get soil samples from the sampling points. Prepare soil suspensions by mixing equal
amounts of soil with distilled water in a beaker (150 ml). Stir the suspension with a glass rod
until soil is completely mixed with water. Wait for the soil particles to settle until a relatively
clear supernate is formed. Get soil pH readings using a pH meter. Be sure the pH meter has been
calibrated before using it.

4. Soil Moisture
Soil moisture content is related to the amount of rainfall, evapotranspiration, drainage and
water-holding capacity of the soil. The relative amount of soil moisture can be determined using
qualitative and quantitative means. A qualitative categorization would be: dry soil (when it is
hard, crumbly and dry to touch); moist soil (when it is pliable and damp to the touch); and wet
soil (when it exudes water when squeezed, leaving the hand muddy). For quantitative
measurement of the percent moisture in the soil, obtain samples from a shallow depth horizon
and seal in separate plastic bags. The bags must be tied up securely and labeled properly for
transport. In the lab room, weigh a clean dry crucible (or beaker, or aluminum dish). Then add
10g of sample, weight it and the container, and oven dry at 105 oC for 24 hours. Remove the
container from the oven using tongs and place it in a desiccator, containing a drying agent such
as anhydrous CaCl2. Wait for the sample and the container to cool at room temperature. After
cooling, weigh the sample with its container. The dry weight of the sample (W d) is computed as
the weight of the container with the oven dried sample (Wo) minus the weight of the container
when empty (Wc):

Wd = W o - W c

The weight of water in the sample is, of course, the difference between the fresh weight and
dry weight. Therefore, the percentage of water in the sample is the weight of the water divided
by the dry weight multiplied by 100 (%).

5. Soil Organic Matter

Soil organic matter is principally composed of humus plus soil organisms and plant roots.
The amount of organic matter in the soil is a major determinant of soil texture, moisture, pH and
nutrients. In the lab room, get the oven-dried samples used in the previous activity on soil
moisture determination. Weigh a clean dry crucible (W c) and fill it with 1-5 grams of oven dried
sample. Weigh again the combined crucible and oven-dried sample. Weigh again the combined
crucible and oven-dried sample (Wo). Heat the soil sample overnight in a muffle furnace set at
4500C. Cool in a desiccator and weigh. Record the weight of the crucible. Record the weight of
the crucible and ignited soil sample as (Wi). Calculate the weight of the ignited soil sample by
subtracting the weight of the crucible (Wi –Wc). The weight lost “on ignition” (Wo – Wi) gives
the organic matter content, which should be expressed as percentage of original dry weight of
sample:
% organic matter = (Wo – Wc) – (Wi – Wc) x 100
(Wo – Wc)
6. Nutrients
Make soil suspension as described in procedure no.3 (soil pH). Determine the presence of
soil nutrients and record (+ / -).

a. Soil calcium
To a test tube containing 10 drops of soil supernate, add 10 drops of solution x (5 grams
ammonium oxalate in 100 ml distilled water). Shake the test tube vigorously to mix the
contents. Varying amounts of calcium is present in the sample if a milky white precipitate
appears after 5 minutes. No color change indicates its absence.
Put a small handful of soil in a crucible and pour excess 10% HCl on it. Observe and listen to
the effervescence. Use table 2 to roughly estimate the amount of calcium carbonate (CaCO 3)
in the soil.
Table 2. Determination of % CaCO3 in the soil sample based on effervescence.
% CaCO3 AUDIBLE EFFECT VISIBLE EFFECT
< 0.1 None None
0.5 Faint None
1.0 Faint – moderate Barely visible
2.0 Distinct, heard away from ear Visible if closely observed
5.0 Easily heard Bubbles up to 3mm, easily seen
10.0 Easily heard Strong effervescence with bubbles up to
7mm

b. Soil nitrates
Add 10 drops of solution Y (0.33 g diphenylamine in 25 ml H2SO4) to a test tube
containing 10 drops of soil supernate. Brown to blue coloration after 5 minutes indicates the
presence of varying amounts of nitrates in the soil sample.

c. Soil phosphorus
Into a test tube containing 10 drops of soil supernate, add 10 drops of solution Z (5g
ammonium molybdate, 50 ml distilled water, 50 ml concentrated HNO3). Add a piece of tin
into the test tube and shake to mix the contents. Gray to deep blue coloration after 5
minutes indicates presence of varying amounts of phosphorus in the soil sample.

d. Soil texture
Classification of soil as to texture can be done by first, feeling the soil whether it is grainy
or sticky. Then, collect soil samples and identify whether it is sandy (particles between 0.5 –
2 mm in diameter, feel gritty or grainy) or clayish (particles less than 0.002 mm, sticky and
may color your hand.
Moisten 10 – 15 cm of soil, knead it, and try to mold it into a ball. Use Table 3 t0 make a
rough classification of the soil into a texture class.

Table 3. Field Key to Soil Texture Class.

SOIL CHARACTERISTICS SOIL TYPE
1 Soil does not remain in a ball when squeezed SAND
1 Soil remains in a ball when squeezed 2
2 Squeeze the ball between your thumb and LOAMY SAND
forefinger, attempting to make a ribbon that you
push up over your finger. Soil does not form a
ribbon.
2 Soil forms a ribbon (may be very short) 3
3 Ribbon extends less than 1 inch before breaking 4
3 Ribbon extends an inch or more before breaking 5
4 Add water to small amount of soil. Soil feels LOAM or SANDY LOAM
slightly gritty.
4 Soil feels smooth. SILT LOAM
5 Soil forms a ribbon that breaks when 1-2 inches 6
long. Cracks if bent into a ring.
5 Soil forms a ribbon longer than 2 inches. Can be 7
bent into a ring without cracking.
6 Add water to small amount of soil. Soil feels at CLAY LOAM or SANDY CLAY
least slightly gritty. LOAM
6 Soil feels smooth. SILTY CLAY LOAM or SILT
7 Add excess water to small amount of soil. Soil CLAY or SANDY CLAY
feels least slightly gritty.
7 Soil feels smooth. SILTY CLAY
C. Biotic Components
1. Plant Species
Observe different plant species found along the 10 –m line in each of the two sampling
areas. Collect 20 leaves (at random) of one dominant species and measure the length and width
in mm. Record your data.
Plot data points as length and width (scatter plots). Determine the y-intercept () and slope
(b) of the graph. The equation for a straight line is
y=a + bx
The point where the line crosses the Y axis is the Y – intercept (). The slope of the line is the
change in the value of y per change in the value of x (Y/X) or b.
A straight (best fit) regression line of the data must pass through the point defined by the
average values of x and y (x,y). It will cross the ordinate at a, where b is the slope.
a = y – bx

y x
The average values are a=∑
n ∑n
−

The best fir straight line minimizes the sum of squares of the departures of the y values from
the line.
Since the line must pass y and x, one can find a if the slope b is known. Hence, b should be
determined using first the following formula:
∑ x∑ y
∑ xy−¿ n
b= ¿
( ∑ x2 ) −¿ ¿ ¿
Once b is found, compute for a using the given formula. You now have a formula for the
best fit line. Illustrate the best fir line for the relationship of leaf length and width. Compare the
two sampling areas (grassy or sunny vs. area with trees or shady).

2. Soil inhabitants
Many small organisms live just beneath and on the soil surface. Collect samples of litter and
place in clear plastic bags. In the laboratory, place a few drops of ether or chloroform in the
bags containing litter. Close the bags tightly. After 5 minutes, empty the contents onto a table
with white paper. Shine a strong light on the litter. Carefully pick all organisms you can find from
the litter using forceps. Put all organisms into a small jar/vial of 70% ethanol. Identify them using
a stereomicroscope. Compare the data from different soil samples.
 ANSWER SHEET

Instructor’s Signature: __________________

Table 1. Readings of physico-chemical parameters taken in different sites.

TRIALS Air Temp. (oC) Soil Temp. (oC) Deeper Soil Temp. (oC) Relative Humidity (%) Soil pH
Sun Shade Sun Shade Sun Shade Sun Shade Sun Shade
1
2
3
MEAN

Table 2. Soil Characteristics.

FACTORS READING/MEASUREMENT
Moisture
Organic Matter
Nutrients
Calcium
Nitrate
Phosphorus
Texture

Table 3. Raw data on leaf sizes of plants from the sunny area.

LEAF LEAF LENGTH (mm) LEAF WIDTH (mm)

XY X2
NUMBER X Y
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
TOTAL
Table 4. Raw data on leaf sizes of plants from the shady area.

LEAF LEAF LENGTH (mm) LEAF WIDTH (mm)

XY X2
NUMBER X Y
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
TOTAL

Leaf
width
(mm)

Leaf length (mm)

Figure 1. Regression line of the leaf data on plants from the sunny area.
 Leaf
width
(mm)

Leaf length (mm)

Figure 2. Regression line of the leaf data on plants from the shady area.

SELF ASSESSMENT:

1. How do the two sampling areas (sunny area vs. shady area) differ from each other?

2. What is the implication of the regression analysis on the leaf characteristics of plants in the two
adjacent areas?

Prepared by:

MARY ANN M. GANZON

SST II - Biology
Checked by:

ROTCHIE GLEN A. FRANCISCO

IS-BIO, Unit Head