The Role of Humidity, Air and Water Temperature and Light

Intensity on Frog Vocalization in the Species Rana clamitans

melanota Located in Michigan at the Grand Valley State
University Arboretum Semi-Permanent Pond.
Brant Morgan, Lindsey Rankin, Victoria Smith & Caleb Westveer
Grand Valley State University is located on the edge of a ravine ecosystem in West Michigan. It
is home to ephemeral and permanent ponds that are located on campus grounds and in the
ravines. For example, the arboretum that is located on campus, houses a semi-permanent pond.
These type of ponds offer unique habitats for amphibians, such as frogs, because they support
sexual reproduction and provide shelter and protection from predators.
The arboretum pond is approximately 15 meters long and 5 meters wide. It is lined with large
rocks and contains the following species composition: water lilies, tall grass, algal mats and other
emergent plants, see Figure 1. This pond is home to only one type of frog, the green frog (Rana
clamitans melanota), see Figure 2.
Figure 1. Habitat in GVSU arboretum pond.

Figure 2. Green Frog located in the arboretum.

Amphibians are ectothermic, which means they do not produce their own body heat and must
obtain it from their external environment (Tracy, 2005). This heat can be gained in the form of
basking in the sun to heat up, or residing in cold waters to cool down. Environmental
temperatures strongly influence locomotion, growth, and reproduction in frogs (Deutsch et al.
2008). Tracy (2005), states that frogs respire through their skin. This process of respiration
results in water loss through the skin; a process that cools the frog much like sweating does for
humans, making it difficult for the frog to maintain the same internal temperature as the ambient

air temperature (Tracy, 2005). Frogs need to reside near water sources to prevent dehydration due
to the process of respiration as well as maintaining their body temperature (Tracy, 2005).
Frogs can emit a variety of different calls such as: terrestrial calls, advertisement calls (to attract
a mate, male only), aggressive calls, distress calls and other calls made during mating (Myers,
2016). Frog vocalizations are energetically costly to produce and body size directly affects the
frequency, intensity and duration of calls produced (Emerson, 2001). The characteristics of these
vocalizations are related to mate selection and are a biotic factor that influence male mating
vocalizations (Gerhardt, 1994). Other biotic factors can influence frog vocalization such as
predation, which has an inverse relationship with frog calls (Ryan et al., 1981). Snakes, birds and
raccoons, commonly found around ponds, may play a role in influencing frog vocalizations, see
Figure 3.
Figure 3. Snake eating tadpole in arboretum pond

The following studies revealed possible atmospheric conditions that affect frog vocalization.
Lingnau (2007) found that the duration of frog calls decreased with an increase in temperature in
the Brazilian torrent frog (Hylodes Heyer) likely because frogs are conserving energy during
periods of high temperatures (Lingnau, 2007). In a new study, frog calling frequency increased
during times of high air temperature (Caldart et al., 2016). This studys findings explained that
because frogs rely on their environment to regulate internal temperatures the increase in
temperatures gave them more energy to vocalize. It was also found that the number of
vocalizations increased when light intensity increased (Caldart et al., 2016). Because light
intensity affects temperature it supports the earlier findings that as the frog's temperature rises it
can produce more vocalizations. It was also found that high relative humidity influences an
increased amount of frog calls (Akmentins, 2014). High humidity helps organisms that require
moist environments like frogs to abstain from dehydration or drying out while conserving energy
(Akmentins, 2014).

Knowing that abiotic factors such as air and water temperature, humidity and light intensity vary
throughout the day and can influence the number of frog vocalizations, we chose to investigate
these factors and their potential influence on the number of frog vocalizations in the arboretum
pond.
We investigated if there was a difference in the mean number of frog vocalizations and abiotic
data throughout the day; specifically comparing morning and evening measurements . We asked
the following questions:
Is there a difference in the mean number of frog vocalizations between morning and evening?
Is there a difference in the mean air temperature between morning and evening?
Is there a difference in the mean water temperatures between morning and evening?
Is there a difference in the mean humidity between morning and evening?
Is there a difference in the mean light intensity between morning and evening?
We hypothesized that air and water temperatures, light intensity and relative humidity would
influence the number of frog vocalizations. We predicted that our evening data would show more
frog vocalizations due to higher air and water temperatures. We also predicted that higher light
intensity would influence a greater number of frog vocalization because of its direct relationship
with temperature. Our final prediction is that when humidity is high we would see an increase in
vocalizations because the frogs would have more energy available because they aren't
dehydrated.
METHODS
We used the Frog and Toad Identification Guide Upper Michigan to determine the species of
frog located in the Grand Valley State University arboretum pond. We determined that the only
species of frog found in the pond was the green frog. The primary distinguishing features of the
green frog are: size (usually ranging between five to ten centimeters long), coloration (green to
brown with some dark, blotchy patches), and a fold of skin that runs the length of the frog from
the eardrum down the back (DNR Wildlife & Habitat. Wildlife Species Amphibians & Reptiles).
Vocalizations of different frog species are distinct and helped us determine that the pond was
occupied by only green frogs. To become familiar with the different vocalization patterns of
frogs commonly found in Michigan ponds, we utilized the University of Michigan Museum of
Zoologys website created by Jones and Hammond 2016.

We chose four time periods throughout the day for observation; sunrise (6:30 AM), mid morning
(10:00 AM), afternoon (5:00 PM) and sunset (8:30 PM). These times were chosen to help
understand what time of day, if any, that frog vocalizations were most abundant. At each of the
chosen times, at least two group members were present to observe and record frog vocalizations
by listening and tallying the number of frog vocalizations heard in each ten minute interval for a
fifty minute observation period. At the end of each fifty minute observation period, the
vocalization data from the two observers was averaged to improve accuracy. We collected data
over a span of three consecutive days (May 23, 24, and 25, 2016).
Before and after each fifty minute observation period, we collected light intensity, air
temperature, water temperature, and relative humidity data. We collected this data from four
different set locations around the pond, as can be seen in Figure 4. We measured light intensity
by using a light meter and recorded this reading in foot-candles. We measured air temperature, in
degrees Celsius, by using a thermometer after it had acclimated for thirty seconds. We measured
water temperature, in degrees Celsius, by placing the thermometer approximately five
centimeters into the pond water and allowing it thirty seconds of acclimation time. Finally, we
measured relative humidity using a hygrometer.
Figure 4. Sampled areas around Arboretum pond.

Image from Google Earth

Following our data collection we conducted T-tests in the program Microsoft Excel to determine
the statistical significance of our findings. To conduct the T-tests, we grouped our data into
morning (6:30 AM and 10:00 AM observations) and evening (5:00 PM and 8:30 PM
observations). The T-tests were used to determine if there was a significance between: frog
vocalizations, air temperature, water temperature, light intensity and humidity, between morning
and evening. Statistical significance was defined as P < 0.05.

RESULTS

The mean, range and standard deviations for our data set is shown in Table 1. These results were
calculated by performing a descriptive statistical analysis on each variable's set of data.
Table 1. Mean, range and standard deviations for frog vocalizations and abiotic factors.
# of Frog
Vocalizations

Air Temp
(C)

Water Temp
(C)

Light Intensity
(Foot-candles)

Relative
Humidity

Mean

50.08

21.20

18.34

5.27

46.04%

Minimum

10.38

13.63

0.00

20.80%

Maximum

123

30,18

24.18

7.75

70.80%

Standard
Deviation

44.33

5.74

3.99

2.54

7.83%

An important aspect of our results is the difference in means between our observation periods.
The average means calculated from our data that was collected during the four fifty minute
observation periods are shown in Table 2. Vocalizations, air and water temperature and light
intensity all follow the same pattern of increasing up until the 5:00-5:50 observation period and
then decreasing after. Relative humidity displays the inverse of that pattern and decreases until
the 5:00-5:50 observation period and then increases after.
Table 2. Vocalization and abiotic measurement means for each fifty minute observation period.
Mean # of
Frog
Vocalizations

Mean Air
Temperature
(C)

Mean Water
Temperature
(C)

Mean Light
Intensity
(Foot-Candles)

Mean %
Relative
Humidity

6:00-6:50 AM

12.5

13.5

3.6

53.50%

10:00-10:50 AM

21.7

22.6

15.7

7.2

45.50%

5:00-5:50 PM

90.3

28.3

22.6

7.6

34.30%

8:30-9:20PM

85.3

21

21.6

2.7

50.70%

The p-values calculated by our t-tests are shown in Table 3. The T-tests showed that frog
vocalizations and water temperature have highly significant differences between the morning and
evening. The T-tests also showed that air temperature was also significant and humidity was just
within the significance range. Light intensity however was shown to be insignificant between the
morning and evening.
Table 3. T-test results for morning and evening data.
P-Value
(Two-tailed T-Test)

Frog Vocalizations

P < 0.001

Air Temperature

P = 0.004

Water Temperature

P < 0.001

Light Intensity

P = 0.840

Relative Humidity

P = 0.046

CONCLUSIONS
The results of our investigation supported our hypothesis that abiotic factors influence the
number of frog vocalizations. Specifically, our prediction that there would be significant
differences in frog vocalizations, air temperature, water temperature and relative humidity
between our morning and evening time periods was supported. Our light intensity prediction,
which stated that a higher light intensity would influence an increase of frog vocalization, was
not supported.
Frog vocalizations and water temperature had the most significant differences between morning
and evening data. We speculate that the reason for the increase in vocalizations in the evening is
due to the warmer air and water temperatures which allows the frog to expend more energy as
was found in the Caldart et al. (2016) study. Figure 1 below shows that when air and water
temperatures are the greatest the number of vocalizations is also the greatest, when air and water
temperatures are the lowest the amount of vocalizations is also the lowest.
The shallow nature of the pond allows for the relatively rapid heating and cooling, due to
environmental factors such as the amount of sun exposure and air temperatures throughout the
day (Butler, 1962). The warmer water temperature helps the green frog regulate its body
temperature which also helps it regulate its energy levels; thus, allowing the frogs to produce
more vocalizations (Tracy, 2005). The expenditure of energy forces frogs to choose periods of
vocalization with a purpose (Emerson, 2001). Our research concluded that warmer air and water
temperatures will make it more favorable for the green frog to vocalize more in the evening
rather than morning.

Figure 1. Frog vocalizations, air temperature and water temperature at selected times.

Research on humidity from Akmentins (2014) showed that frog vocalization increased when
humidity was high. Our data found that there was a significant difference in humidity between
morning and evening; however, as stated in the results section there is an inverse relationship
between humidity and frog vocalizations. In order to support or disprove our prediction of frog
vocalization vs. relative humidity, we would have to record more data over a longer period of
time.
Light intensity was shown to not have a significant difference between the morning and evening.
Our research from Caldart et al. (2016) found that light intensity did influence frog vocalizations;
our data suggested otherwise. This can be explained by understanding the relationship between
light intensity and air/water temperature; both variables which we did find to have an influence
on frog vocalizations. When the sun sets the light intensity plummets and the temperature of air
begins to decrease quickly; the water temperature however remains relatively stable due to the
high specific heat capacity of water (Perlman, 2016). This suggests that light intensity does not
influence frog vocalization but that it affects the temperature of the air and water which do affect
vocalizations.
Our investigation could be improved if we had access to equipment that allowed for continuous
gathering of data. This would allow us to potentially establish a correlational relationship
between frog vocalization and the air and water temperature along with the relative humidity.
The disturbances, which consisted of us making noises while walking around the pond when
collecting data, lawn maintenance equipment of GVSU near and away from the pond, and other
people visiting the pond may have influenced the frog activity. Being able to record vocalizations

without disturbing the frogs would help eliminate other factors that may be affecting their
vocalizations.
Our results support our prediction that air and water temperatures influence frog vocalization.
Further testing is needed to support or disprove our prediction that high humidity influences frog
vocalizations and our prediction that light intensity influences frog vocalization was disproven.
Even though our study could be improved for more accurate results it did yield conclusions that
we feel will be repeatable with further testing.

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