Bollig, Cataggatan, King, and Rampone 1

The Effects of Moisture in the Turn Alternation Behavior of the Species Porcellio

Alec Bollig, Paolo Cataggatan, Mitchell King, and Kate Rampage Rampone
Jake Griffin and Jacob Usinowicz, Section 14

Introduction

Turn alternation behavior has been observed in several species during many different

experiments, but the exact causes and benefits are still a topic of debate (Hughes 1986). This

concept describes the tendency of certain organisms to change the direction of their successive

turns. In other words, immediately following a left turn, many species show a preference to turn

to the right at the next intersection and vice versa (Harvey and Bovell 2006). This phenomenon

has been recorded in paramecium (Harvey and Bovell 2006), earwigs, earthworms, millipedes,

woodlice (Hughes 1986), pillbugs (Ono and Takagi 2006), and many other species in tests

conducted by numerous scientists. As was mentioned, there has been some controversy regarding

the motives for turn alternation behavior in these organisms, but there are several hypotheses.

One proposal is that alternating turns serve to maintain a relatively linear route (Hughes 1986).

The idea is that repeating the same turn direction would cause a circular course with little net

movement. However, by changing directions at each intersection, an organism is able to move in

a roughly direct path to food, safety, or some other motivator (Hughes 2007). Other theories have

also been offered, and several variables have been tested in relation to turn alternation. The size

of the organisms, the angle of their turn, and the distance between turns are just a few of the

things that have been studied (Hughes 1986; Hughes 2007). The purpose of this specific

experiment is not to further support the well documented notion of turn alternation behavior, but

to study the affect that moisture may have on its rate of occurrence.
 Bollig, Cataggatan, King, and Rampone 2

The organisms used in these tests were of the species Porcellio,which are better known as

woodlice or sowbugs. These arthropods are one of the few species of terrestrial crustaceans, but

they still require a certain level of moisture for survival. In contrast to many other species of

arthropods, the exoskeletons of woodlice do not have a waxy outer layer. The absence of this

coating leave sowbugs very susceptible to water loss, or desiccation. Drying out can prove fatal

because respiration cannot occur without moisture, and despite their pseudotracheae being more

suited for dry conditions, the gills of sowbugs are very dependent on water (Raham 1986).

Woodlice are ideal for the study of the effects of moisture on turn alternation behavior because

their close relative, the pillbug (Ono and Takagi 2006), and they themselves have been shown to

demonstrate turn alternations on multiple occasions (Hughes 1986; Hughes 2007). Additionally,

past studies have implied that moisture in the air has the effect of raising the rate at which

terrestrial isopods alternate their turns (Moriyama 1999).

Given the theory of turn alternation behavior maintaining direct paths and the preference

of woodlice to be in a fairly moist environment, it was expected that the sowbugs in this study

would demonstrate fewer turn alternations in a more moist environment. This hypothesis was

founded on the concept that the woodlice would reduce turn alternations in order to remain in

closer proximity to the essential water source. Conversely, if they were to alternate their turns,

they would eventually end relatively far from the damp area. To study the effects that moisture

may have on the turn alternation rate of woodlice, specimens were be placed in separate wet and

dry mazes where their behavior was observed closely.

 Bollig, Cataggatan, King, and Rampone 3

Methods

Approximately forty Porcellio specimens of lengths varying from ten millimeters to

sixteen millimeters were used for testing, and no particular attention was paid to the sex or age of

the animals. When they were not navigating the mazes, the woodlice were kept in one of two

aerated plastic containers along with a slightly damp paper towel. Every sowbug started in the

same container, but to ensure that no one subject was overworked, they were placed in the other

following their first run. After each specimen was used once, the process repeated, and this same

pattern was followed until sufficient data had been collected. The subjects all experienced the

same light conditions as the others before them, and every trial was done at a normal room

temperature of about eighteen degrees Celsius.

Three separate T-mazes constructed from Legos and the occasional Mega Blok were used

during the experiment. These mazes were placed on top of paper towels to provide a disposable

surface that could be changed before each run to erase any chemical signals left behind by the

woodlice. Every block used in making the mazes was red, which served to eliminate any

preference in color, tint, or shade that sowbugs may have. The lengths of the paths were also held

constant at roughly four centimeters in each of the mazes and trials to reduce the effects that

short-term memory loss could have had on the woodlice. Additionally, the width of every track

was about eight millimeters which ensured that the specimens were within close contact with

both side walls. This helped to decrease the likelihood that the animals would simply follow the

surface of the blocks that they were touching. Two of the mazes consisted of a single forced turn

followed by an intersection that allowed the specimen to choose between a right or left turn.

However, these two mazes were not identical as one forced a left turn first while the other
 Bollig, Cataggatan, King, and Rampone 4

required an initial turn to the right. The third maze was a bit more complicated and gave the

subject four separate intersections with left and right options. None of the turns in any of the

mazes had any significant incentives, such as food or Pringles, that would have affected the

decisions of the subjects.

Before the experimental data was recorded, a control set was collected for comparative

purposes. Since the intent of this experiment was to study what effects, if any, moisture has on

the turn alternation behaviors of woodlice, the control group performed their test runs on dry

paper towels. Twenty trials were run with each maze, but the maze with no forced turns allowed

for three possible turn alternations, so three times as much data was collected for this type of run.

In the two mazes with forced turns, the sowbugs were placed at the start where they proceeded

forward until they reached the first turn. A turn alternation was recorded if the subject turned in

the opposite direction of the forced turn at the intersection that followed. If the specimen did not

exhibit a turn alternation, it was noted by stating that no alternation behavior was demonstrated.

Special attention was paid in the multiple path maze because the animal could make three, two,

one, or zero turn alternations in any given order. As in the other two mazes, the animal was

placed at a start, but instead of simply paying attention to one turn, the exact path of the subject

was written down. Afterwards, the number of turn alternations was determined and represented

by a fraction out of the three alternations that were possible. When a specimen was unwilling to

navigate the maze on its own, a fine bristled paintbrush was used to coax them into moving. The

same processes and number of trials described above were done a second time with damp paper

towels instead of dry ones. Lightly spraying the paper towels with a water bottle served to create

the moist environment that was being tested.

 Bollig, Cataggatan, King, and Rampone 5

After the necessary data was collected, it was analyzed using two different statistical

methods. One approach was to calculate the percent of turn alternations in the three mazes for

both wet and dry environments. The standard deviation and standard error of these percentages

were then figured to determine if the findings were of significance. The second method was a

chi-square test that used the information from the dry runs as the expected values since the study

was concerned with the effects of moisture. Wet and dry environments were the only two

categories, so there was just one degree of freedom in this scenario. The null hypothesis in this

case was that introducing moisture would have no effect on the rate of turn alternation.

INSERT MAZE DIAGRAMS

Results

The data from the two forced turn mazes were combined into one set because there was

no evident difference between an initial right or left turn in either series of tests. The subjects

showed turn alternation behavior about 70% of the time in the dry runs with a standard error of

4.9% (Figure 1). When moisture was introduced, the mean percentage of turn alternations in the

woodlice rose to 81.7% with a standard error of 3.4% (Figure 1). The chi-square test that was run

provided a value of 2.48 with one degree of freedom, which yielded a probability value of 0.115.
100
90
80
Figure 1. A comparison of the percent of turn alternations
70
Turn Alternation %

observed in the dry mazes opposed to the moist mazes.

60 The percentages were found by dividing the number of
50 observed turn alternations in each maze by the number of
40 possible alternations. The error bars represent the standard
error for each of the mean percents.
30
20
10
0
Dry Moist
Environment
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Discussion

The results of this experiment were somewhat inconclusive because the statistical models

expressed varied significance. However, the hypothesis that was being tested was strongly

contradicted. Not only was there no substantial data to support the hypothesis, but the

information that was collected actually suggested that moisture has the opposite effect on turn

alternation behavior. The sowbugs alternated their turns even more in the moist environment

compared to the dry one. In terms of percentage, there was a minimum of a 3.4% increase in

alternation rate when standard error is considered. This may not seem like much, but the lack of

overlap in the error suggests that this difference has some significance. If the error was examined

from the opposite extreme, a formidable 20% increase in turn alternation rate would be shown in

a moist environment compared to a dry one. The chi-square test did not provide the same results,

though. With just one degree of freedom, a value of 2.48 and a probability of 0.115 are just out of

the significant range. The discrepancy between the mean percent of turn alternations and the chi-

square test leave drawing conclusions from the results somewhat difficult. Despite this

inconsistency, the hypothesis that moisture will decrease turn alternation behavior in woodlice

was sufficiently rejected.

The lack of statistical evidence supporting the given hypothesis in this experiment

suggests that a flaw in biological rationale was made. Another study concerning turn alternation

behavior in terrestrial isopods implied that fleeing from a threatening situation caused an

increase in turn alternation rate (Ono and Takagi 2006). The subjects in the current test were

abruptly displaced before navigating the mazes, and those that would not comply were lightly

urged forward. The woodlice may have increased their turn alternation to cope with their new
 Bollig, Cataggatan, King, and Rampone 7

surroundings and the somewhat intrusive paintbrush strokes, ignoring the moisture in order to

escape a threat. A more patient approach to testing may have reduced the effects of stress on the

behavior of the woodlice, but time was a limited resource when this study was conducted.

The anxiety of the animals may not have been accounted for, but several measures were

taken to reduce the effects of variables other than moisture. Even so, the experiment did have

other flaws as well. While the paper towels beneath the mazes were changed before each new

trial, there was no thought given to the possible chemical trail left on the blocks themselves. This

means that the woodlice may have been following the scent or some other stimuli left on the

maze by an earlier trial. Another scenario that was overlooked pertains to the memory capacity of

the specimens. The tracks were all kept at the same, relatively short length, but nothing was done

to control the time of each trial. One subject could finish the course in just a few seconds, but

another could stop after the first turn and wait for a while, giving it ample time to forget what

had just occurred. Without recollection of the first turn, the second turn would be purely random

and would not be representative of a turn alternation. Light also could have effected the woodlice

even though the intensity was never changed. Sowbugs have shown a negative reaction to light

in studies done on their behavior (Warburg 1968), so they may have turned based on which side

had more shade. Since the light was not applied directly from above, shadows may have been

cast by the maze walls. This possibility is supported by the observation that many specimens

stopped immediately after turning, which suggests that they were content with the level of

darkness in certain locations. Any of these missteps could have skewed the data that was

collected during this experiment, but the overwhelming evidence against the presented

hypothesis is hard to discredit. However, if this study were to be repeated in an attempt to

 Bollig, Cataggatan, King, and Rampone 8

support that moisture increases the turn alternation rate of woodlice, these flaws would need to

be addressed to provide for the most accurate, uninfluenced information.

As was just proposed, another series of tests could be run using the opposite hypothesis

of this experiment. A second option would be to keep the same hypothesis but to reduce the

threatening dislocation of the specimens that may have interfered with the results. In either case,

it would be helpful to use more similar maze types when observing forced turn behavior and free

turns. Having two mazes with a single forced turn and one option and a third with four options

worked, but it made things slightly more difficult than they had to be. The raw data collected

from these two different methodologies needed to be manipulated in order to be comparable.

Building a free turn maze with only two intersections would make analyzing data much easier

because only one turn alternation would be possible in each maze.

Ultimately, no strong evidence was found to support the hypothesis that moisture has a

significant effect on turn alternation behaviors in sowbugs. The exact reasoning for turn

alternation remains an elusive and argued piece of information that is certainly influenced by

several variables that can vary between species. Only by formulating more hypotheses and

conducting more studies can the phenomenon of turn alternation be understood more clearly.

Literature Cited

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Learning & Behavior. 34(4): 361-365.

Hughes, R.N., 1986. Mechanisms for turn alternation in four

invertebrate species. Behavioural Processes. 14: 89-103.

 Bollig, Cataggatan, King, and Rampone 9

Hughes, R.N. 2008. An intra-species demonstration of the independence of distance and

time in turn alternation of the terrestrial isopod, porcellio scaber. Behavioural

Processes. 78: 38-43.

Moriyama, T. 1999. Decision making and turn alternation in pill bugs (armidillidium

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Ono, T. and Takagi, Y. 2006. Turn alternation of the pillbug armadillidium vulgar and its

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Raham, Gary. 1986. Pill Bug Biology: A Spider's Spinach, but a Biologist's Delight. The

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Warburg, Michael R. 1968. Behavioral Adaptations of Terrestrial Isopods. American

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