Observation and Measurement of Behavior of Mus Musculus

8/2/2019 Observation and Measurement of Behavior of Mus Musculus

1/17

Observation and Measurement of Behavior ofMus musculus1

Marie Arenbi Carillanes

Eunice Marie Jaen

Arriane Mae Isla

Ivy Madrid

Paulo Miguel Kim

1A scientific paper submitted in partial fulfillment to the requirements in Animal Behavior Laboratory under Mr.

Pablo Ocampo, 1st

sem, 2011-2012.


2/17

INTRODUCTION

In a purely biological perspective,behavior is defined as all observable muscular and

secretory responses to changes in an animals internal or external environment (Grier, 1984).

However, such a definition is far too broad. Behavior can be encompassed by physiology,psychology and sociology. But it can also be tackled in the field of zoology. In its simplest form,

one can consider behavior as the collection of observable and recognizable actions of animals.

Any study of animal behavior always starts at the same line. By far it is the most important

yet also one of the most overlooked. A thorough knowledge of the target specimen being studied

is a critical necessity for any behavioral studies. It ultimately will lead to a proper citation and

formulation of a special behavioral set unique to a species, a list known as an ethogram.

An ethogram is a repertoire composed of sequences of behavioral patterns. Each pattern in

turn is composed ofethons, the smallest unit in the study used to denote the tiniest action of

behavior which makes up the whole (Ibid.).

In dissecting the total behavior displayed by an animal, one can derive the ethons and

attribute the correlations of each in forming possible conclusions to their existence.

Some behavior of animals are innate in nature, a necessity for survival (Breland, 1966). But

because behavior is variable, it cannot be expected that the different animals will display the

same behavior. Thus, a large amount of time is used just to document the list of behavior of a

single animal and a greater amount of patience is employed to ensure that the list of behavior is

as close to the animals.

However, the number of acts that can be observed in an animals repertoire is dependent in

three factors. First, one must consider the number of acts that an animal possesses. Second, the

rarity of a certain act within a bout must also be considered, whether or not observers will be

able to ascertain such behavior. Lastly, one must consider the duration of observation. The

longer one observes the higher probability of seeing the rarer acts.

Behavior can be grouped accordingly in a logical, natural categories based on their function.

They can also be illustrated according to the measurement of the continuous aspects of an

animals appearance (i.e. posture, angle or position on limbs) (Grier, 1984).

Statistics provide studies in behavior the necessary tools in order to understand therelationship of the observations made. Descriptive statistics aid in describing the characteristics

of a set of numbers. Inferential statistics on the other hand, is used when one infers, with the aid

of assumptions and statistical tests like analysis of variance, how groups of data relates to each

other and if whether there is an inherent order or variability. Between the two, inferential

statistics is more highly used, the former often regarded as existing under the auspices of random


3/17

change rather than by any underlying effect of interest.

Often times, behavior is taken is as part of a multiple whole rather than as a single entity.

Among the common categories of analyzing multiple behavior data are the comparison of the

frequency of behavior among different rest subjects, the analysis of the interaction of behaviorbetween two or more individuals and the analysis of the sequence of behavior through time.

Statistical analysis is further subdivided into two types: parametric analysis, where the

assumption that a population is normally distributed and variance is assumes homogenous and

non parametric, where it is not necessary for a population to be normally distributed or for

variance to be homogenous(Changing Minds, 2011).

The common house mouse (Mus musculus) is a common test animal in the study of behavior.

It exhibits a variety of behavior and is easy to monitor as well as manipulate. It is an efficient

subject particularly for observing behavioral patterns and for detecting ethons within series ofcomplex acts. Grooming behavior is among the most observed acts among mice and an ideal

study for sequential behavior.

The objectives of the experiment is to study the grooming behavior of animals, specifically

Mus musculus, Furthermore, the experiment will ascertain the grooming behavior ofMus

musculus under stress and to determine the significance of the behavioral pattern and its subunit

suingthe following measures: duration, interval, latency and sequence. In the study, the

principles of observation, description and quantification of behavior will be performed and

evaluated.

This study was conducted at A-125, Institute of Biological Sciences, UPLB on August 2011.


4/17

METHODOLOGY

In ascertaining the grooming behavior of mice, an observation chamber set up was assembled.

Three aquarium tanks were setup. Fluorescent light was wrapped in red plastic to mask the light.

The sides of the aquarium were covered to delimit the red light within. Three mice (an individual,

a male and a female) were produced. The individual test mouse was coated in flour inside aplastic bag and allowed to acclimatize in the observation chamber for 5 minutes. The grooming

behavior of the mouse was complied and labeled into specific codes for later use. Observation

was made for 15 minutes.

The test mouse was redusted and reintroduced to the chamber. The procedure was repeated for

another 20 minutes. The grooming acts were noted using the formulated codes and the actual

time until the end of grooming was recorder in seconds. All data and observations was tabulated

(Table 2).

The male and female mice were tested separately. Each test mice was sprayed with copious

amount of water and placed in the chamber. Using the previously formulated codes, all grooming

acts displayed were noted in a sequence for 30 minutes at a one minute interval. All data and

observations were tabulated (Table 3).

The mean of the duration, interval and frequency of the first experiments were computed and

tabulated. Similarly, the mean frequencies of the male and female mice for the second

experiment were also computed.

All computed measures were pooled. The Kruskall-Wallis Single Factor ANOVA was

applied. Results were analyzed and predominant sequences were established from the data.


5/17

RESULTS AND DISCUSSIONS

The data was subdivided into two main experiments. An initial set up was established to

observe grooming subunits as well as to establish a sequence code for later use (Table 1). The

first experiment involved the use of an individual mouse placed into a condition to elicit its

grooming behavior, the condition here is the use of flour as a stimulus. Two sets were made, thefirst to ascertain the categories and the subunits of its behavior based from observation. The

second set was implemented to establish the sequence of the behavior being displaced under 15

minutes and form a conclusion based on the resulting frequency, interval and duration of the

behavior (Table 4., Table 5., Table 6.)

The second experiment involved two specimens, a male and female mouse. Similarly to the

previous, the experiments involved subjecting the specimens into specific conditions which

would elicit their grooming behavior. However, unlike them former, water was used instead to

ascertain the resulting behavior. The previously established categories and subunits were also

applied in the experiment as the same procedures. However, only the frequencies of thecategories were taken. (Table 7., Table 8. )

The results of the first experiment showed that, of the four categories of grooming (Head,

Body,Coxal and Leg), the longest duration as well as interval for the two specimens was for

body grooming (8.36 and 8.2 seconds respectively) for each bout.

On the other hand, the most frequency grooming category exhibited was seen in leg grooming,

with a mean total of 26.5.

The results of the second experiment, on the other hand, showed that for males, there was

higher average frequency of leg grooming in contrast to the other categories as well as those of

the female (34.25).

The female specimens, however, exhibited a higher average frequency of head grooming

against the other categories (27.25).

In order to establish the significance of the data set, a non-parametric statistical tool was

employed. Instead of using single factor ANOVA, Kruskall-Wallis single factor ANOVA was

used. The Kruskall-Wallis one-way analysis of variance uses ranks of the measurements instead

of the actual measurements of variabilities of the populations being compared. It is an extremely

useful test on deciding whether kindependent from different populations. Furthermore, the testassumes that the variable under study has an underlying continuous distribution (Table 9).

Using the Kruskall-Wallis single factor ANOVA, the following results were obtained:

1. With H values of 3.17, 3.17 and 4.17 respectively, it was concluded that there was nostatistical significant differences among the durations, intervals and frequencies of the

four grooming behaviors in the two mice specimens.


6/17

2. With H value of 5.41, it was ascertained that there was no significant, statisticaldifferences between the frequencies of the four grooming habits in the four male mice

specimens.

3. With a H value of 7.18, it was determined that there was no significant statisticaldifferences between the frequencies of the four grooming habits in the four female mice

specimens.

From the results of the experiments, it can be established that there is not significant

differences in the different grooming habits of the mice. As such, although some grooming

categories and subunits were exhibited more within a single bout than others, there is no

statistical evidence that there is a constant, established pattern being followed.

However, from the results of the experiment, it was found that in terms of frequency, leg

grooming and its associated subunits was potentially higher than those of the other categories.

But in terms of both duration and interval, body grooming was exhibited longer and within

greater intervals. It will also be noted that head grooming closely followed both of the previous,acting as an intermediate in terms of frequency, duration and interval.

While it cannot be sufficiently established from the results, it may postulated that the after

mentioned three categories of grooming were more exhibited in a given bout. Coxal grooming

proved to be the least exhibited and in fact has the lowest values in terms of the three

parameters.

Anxiety and stress often plays a major part in grooming amongst many mammals, especially

among mice. Depending on the level of stress, grooming may be erratic, passive or robust

(Kyzar et al., 2011; Animal Behavior Society, 2011).

In mice, the more anxiety experienced, the more vigorous grooming is. With the setup of the

experiment, stress is reduced by a combination of acclimatization and the red light that prevents

the specimens from detecting the observers.

However, stress was added in order to elicit grooming responses, namely subjection to flour

and water, as seen in the first and second experiment respectively. Thus, it was expected that the

mice were to exhibit vigorous grooming behavior (Kyzar et al., 2011).

In the flour stimulus experiment, although leg grooming was more frequently exhibited at about, body grooming was performed at a longer duration as well as in greater interval. As was

previously mentioned, head grooming was considered at an intermediate position between the

two. Coxal grooming was not emphasized nor displayed recurrently.

In the water stimulus experiment, while the duration and interval were not noted, the

frequency of the male and female mouse were ascertained. The male mouse showed an higher


7/17

average frequency of leg grooming (34.25), followed by head grooming (27.25) whereas the

female showed a higher average frequency of head grooming (27.25), followed by leg grooming

(24.5). Coxal grooming was low in terms of frequency, for both male and female mice.

Of the four grooming categories, the head and body grooming were more emphasized by

mice due to the inherent consequences associated with keeping the two regions free fromirritants. The head region involves many of the senses relied upon by mice in order to cope with

its environment. Likewise, the body is necessary in order to maintain coordination among mice,

an important trait for survival.

On another note, the leg region, though only frequent in duration in the experiment, also is

an important area which must be maintained by mice, being its essential tool for

locomotion.

If it is thus analyzed, the head region houses the vital sense organs like the eyes, ears and

nose. These sense organs are used by mice in order to survey their environment for any subtle

changes that may signal for security as well as for detecting food. Irritants that impede the

functions of these senses must be removed to maintain optimum use of these senses. On the

other hand, once the brain processes the stimuli, it must act depending on the significance of the

message, whether to maintain its position or to escape. Likewise, the body as well as the legs

must be groomed to avoid critical mistakes in its behavioral responses. Irritants that block the

optimum use of the body and limbs could prove a fatal mistake for many a species.

However, in mice, it was observed that the coxal grooming was the least emphasized

grooming category and the least observed in a particular bout. It may be postulated that coxal

region, a region whose usual function is to maintain gait during locomotion, is least displayed

because the tail poses no immediate role in behavioral mechanisms like escape and detection.

Thus, even in a passive environment where the levels of threat are low, the mice specimens

perform coxal grooming less frequently as compared to grooming the other regions.

Lastly, it must be noted that the tail, unlike most of the other body regions, has relatively

sparse fur and thus least likely to be affected by irritants like dirt and parasites.

There is a distinguishable sequence which maybe observed within the grooming acts. This

sequence is best seen in the second experiment where the male and female mice were involved.

As can be seen in Table. 2 and 3, there is a complex number of apparent sequences which maybe observed. However, segments of the major sequence of grooming behavior can still be noted.

It will be noted that majority, if not all, of the grooming acts per one minute were initiated by

head grooming. Although no true pattern could properly be established, the head grooming

subunits began with licking or scratching different to successive parts of the head.


8/17

After head grooming, body or leg grooming followed. However, between the two, leg

grooming proceeded first before body grooming, though instances arose where in the latter

appeared in the absence of the former. The grooming step usually proceeded from the leg to the

sides of the body, gradually progressing ventrally until the anal-genital regions was reaches.

Finally, observations yielded that a singular pause was usually performed before proceeding

to coxal grooming.

In general, a grooming sequence could be established: Head-Leg-Body-Coxal.

However, it will be noted that grooming of the coxal region was infrequent and thus, there

were only few instances where in the entire sequence were actually observed.

Other than that, there were frequent recurrent fragments of this major detectable sequence.

Each fragment follows the established sequence closely and with little to no deviation.

The after mentioned grooming sequence that was established by the experiment is common to

all rodents. All rodents engage in a number of self-grooming activities in order to keep the fur

and the skin clean. The form in which these sequences as displayed may differ from one rodent

to another but the basic pattern is retained. Other times, the organization varies from veryloosely to very stylized though always in an identical fashion. This pattern is an extensive

grooming activity known as cephalocaudal grooming. This grooming sequence, begins from the

face (head region) and gradually covers up to the flanks (the leg and body region) and finally to

tail (coxal region). Grooming, however, may be interrupted from between points in the

sequence, most notably the abrupt pause before tail grooming (Rat Behavior, 2008; 2011).


9/17

SUMMARY AND COCNLUSION

The observation and measurement of behavior of the albino variant house mouse (Mus

musculus) has been determined.

The compilations of a behavioral catalogue for the specimens as well as the dissection of the

individual bouts have yielded four major grooming categories (Head, Leg, Body and Coxal),

each of which were further subdivided into subunits.

Results of the initial experiment on individual test mice showed that leg grooming , with a

mean frequency of 34.25, showed the highest frequency displayed whereas the longest duration

and the greatest interval was shown in body grooming (8.36 and 8.2 seconds respectively).

Results of the second experiment showed that the highest average frequency of grooming

behavior in males was under leg grooming (34.25) whereas the highest average frequanecy for

female mice was under head grooming (27.25).

Using Kruskall-Wallis Single Factor ANOVA, it was determined that there were no

significant statistical differences between the duration, interval and frequency of the four

grooming acts of the individual mouse in the flour test. Similarly, there were no significant

statistical differences between the frequency of the four grooming behavior of both the male and

female mice in the water test.

Thus, there is no direct correlation that a grooming act on a certain body part is exhibited

more than the others.

Lastly, a detectable sequence was established from the resultant data. A pattern of Head-

Leg-Body-Coxal grooming was determined to persist in any series of behavior displayed by the

test mice, a main sequence which is followed whether whole or in fragments. Furthermore, the

cephalocaudal grooming displayed was loosely organized, the intervals often interrupted, thus

giving rise to the fragments of the main sequence.

The measurement of behavior is mediated by the careful use of observation as well as

statistical analysis in order to ascertain significance. In the study of ethograms, the initial step is

to determine the smallest identifiable units of behavior by dissecting the observable behavior of

an animal. Behavioral patterns can be determined after following the complex sequences formed

from these units.

In the case of the test mice, the smallest units that make up the entire behavioral pattern are

ultimately responsible for the overall makeup of the sequence. By determining parameters such

as duration, interval, latency and frequency, it is possible to determine a possible base sequence

of behavior.


10/17

REFERENCES

Animal Behavior Society.Ethogram of Mice. . Accessed August 2011.

Breland, K. and M. 1966.Animal behavior. New York: The Macmillan Company.

Changing Minds. Parametric vs. non-parametric tests.

. Accessed August 2011.

Grier, J. W. 1984.Biology of animal behavior. St. Loius, Mo.: Times Mirror/Mosby

College Publishing.

Kyzar, E., Gaikwad, S., Roth, A., Green, J., Pham, M., Stewart, A., Loang, Y., Kobla, V.

and Kallueff, A. 2011. Towards high-throughput phenotyping of complex patterned behaviors in

rodents: Focus on mouse self-grooming. New Orleans: Elsevier B.V.

Rat Behavior. 2008.Rat Behavior and Biology. .

Acessed September 2011.

---------------2011.Glossary of rat behavior terms. .

Acessed September 2011.

Siegel, S. 1956.Non-parametric statistics for the behavioral sciences. USA: McGraw-Hill

Book Co.
http://www.animalbehavior.org/ABSEducation/laboratory-exercises-in-animal-behavior/laboratory-exercises-in-animal-behavior-ethogramshttp://www.animalbehavior.org/ABSEducation/laboratory-exercises-in-animal-behavior/laboratory-exercises-in-animal-behavior-ethogramshttp://www.animalbehavior.org/ABSEducation/laboratory-exercises-in-animal-behavior/laboratory-exercises-in-animal-behavior-ethogramshttp://www.animalbehavior.org/ABSEducation/laboratory-exercises-in-animal-behavior/laboratory-exercises-in-animal-behavior-ethogramshttp://changingminds.org/explanations/research/analysis/parametric_non-parametric.htmhttp://changingminds.org/explanations/research/analysis/parametric_non-parametric.htmhttp://changingminds.org/explanations/research/analysis/parametric_non-parametric.htmhttp://www.ratbehavior.org/norway_rat_ethogram.htmhttp://www.ratbehavior.org/norway_rat_ethogram.htmhttp://www.ratbehavior.org/norway_rat_ethogram.htmhttp://www.ratbehavior.org/Glossary.htmhttp://www.ratbehavior.org/Glossary.htmhttp://www.ratbehavior.org/Glossary.htmhttp://www.ratbehavior.org/norway_rat_ethogram.htmhttp://changingminds.org/explanations/research/analysis/parametric_non-parametric.htmhttp://www.animalbehavior.org/ABSEducation/laboratory-exercises-in-animal-behavior/laboratory-exercises-in-animal-behavior-ethogramshttp://www.animalbehavior.org/ABSEducation/laboratory-exercises-in-animal-behavior/laboratory-exercises-in-animal-behavior-ethograms


11/17

APPENDIX

Table 1. Catalog listing of grooming behavior.

Grooming Act Subunits Codes

Leg

Lick arm Scratch arm LLA LSA

Lick leg Scratch leg LLL LSL

Lick feet Scratch feet LLF LSF

Head

Lick ear Scratch ear HLE HSE

Lick nose Scratch nose HLN HSN

Lick cheek Scratch cheek HLC HSC

Lick head Scratch head HLH HSH

Body

Lick back Scratch back BLB BSBLick stomach Scratch stomach BLS BSS

Lick testicle Scratch testicle BLT BST

Coxal

Lick base tail Scratch base tail CLB CSB

Lick middle tail Scratch middle tail CLM CSM

Lick tip of tail Scratch tip of tail CLT CST


12/17

Table 2. The sequence of grooming acts in terms of duration and interval.

Succession of Grooming Acts Gooming Act (coded) Beginning time (s) End time (s) Duration (s)

1 HLH 5:05 5:05 15

2 LLF 5:06 5:06 2

3 HLH 5:06 5:06 3

4 HLH 5:06 5:06 2

5 HSN 5:06 5:07 3

6 LSF 5:08 5:08 2

7 LSF 5:08 5:08 2

8 BLS 5:08 5:09 3

9 HLH 5:09 5:09 6

10 LLF 5:09 5:09 5

11 HLH 5:09 5:09 3

12 HLH 5:09 5:10 2

13 LLF 5:10 5:10 3

14 LLF 5:10 5:10 4

15 BLB 5:10 5:10 7

16 LSF 5:10 5:10 3

17 HLH 5:10 5:10 3

18 BLT 5:10 5:10 5

19 LLF 5:11 5:11 10

20 BLS 5:11 5:11 8

21 BLS 5:11 5:12 4

22 BLB 5:12 5:12 16

23 LLF 5:12 5:12 11

24 HLN 5:13 5:13 3

25 LLF 5:13 5:13 7

26 LSF 5:13 5:13 4

27 LLF 5:13 5:14 10

28 HLC 5:14 5:14 3

29 BLB 5:14 5:15 9

30 HSN 5:16 5:16 15

31 BLS 5:16 5:16 14

32 LLF 5:16 5:16 5


13/17

33 HLH 5:16 5:16 2

34 BLS 5:16 5:17 12

35 BLT 5:17 5:17 7

36 HLH 5:17 5:17 4

37 BLT 5:17 5:17 5

38 HLH 5:17 5:18 6

39 BLB 5:18 5:18 13

40 BSB 5:18 5:18 12

41 LLF 5:18 5:18 7

42 LSF 5:19 5:19 5

43 BLS 5:19 5:19 9

44 HSN 5:19 5:19 3

45 LLF 5:19 5:20 9

46 HSE 5:20 5:20 3

47 HSE 5:20 5:20 4

48 HSN 5:20 5:20 2

49 HSN 5:20 5:20 3

50 HSN 5:20 5:21 2

51 BLS 5:21 5:21 8

52 HLH 5:21 5:22 4

53 HLH 5:22 5:22 5

54 HSC 5:22 5:22 2

55 BLS 5:22 5:23 14

56 LLF 5:23 5:23 8

57 HLN 5:23 5:23 558 LLF 5:24 5:25 6

59 LLF 5:25 5:25 12


14/17

Table 3. The sequence of grooming acts of male and female mice.

Animal 1 (male) Animal 2 (Female)

No. Grooming Acts (Coded)

No. Grooming Act (Coded)

1HSM, HLN, HLH, HSC, HSE,

LSF,LLF 1 HSN

2 HSC 2 HSN, HSE, LSF, BSB

3 HSN, LLF 3 HSE, HSN, HSC, LSF, LSF, BSB, BLB

4 HLH, LLF, BLS 4 HSE, HSC, HSE, LLF, LLF, BSB

5 HLH,HSN, HLC, LLF, BLS 5 HSF,BLS, BLB,CLB

6 HSB, LLF, BLB 6 -

7 HSE, LLF, LLH, BLB 7 -

8 HSE, HSN, HSC, LLF, BLB,BLT 8 HSF, LSF

9 HLH, HSN, BLB, 9 -

10 HSN, LLF, BLS 10 HSN

11 HSN, HLH 11 -

12 HLH, HSE, BLS, BLB 12 -

13 HLH, LSF, BLB, CEB 13 HSF

14 HSN, HLH, BLB 14 -

15 HLH, HSE, BLB 15 -16 HLH, HSE, BLB 16 HSE, HSN, HSC, HLE, LSF,LLF

17 HSN, LLF, BSB, CLB 17

HLN, HLE, HSE, HSE, HSN, LLF, BLG, BLS,

BLB

18 HSC, HSN, HLOH, LLF, BLS, BLB 18 HLH, HLN, BLB, BLG, BLS

19 HLH, BLT, BLB 19 HLE, HLC, HLH, HLN, LLF, BLB, BSB, CLB

20 HSE, HSN,LLH, BLB, CLB 20 HSE, HSN, LLH, LLF, BLB, BSB, BLG, BLB

21 HSE, HLN, HLH, HSE, HSC 21 HLH, HLN, HSE, HSC, BLS, BLG, BLB

22 HSE, HSN, HSC, BLB, BLS 22 HLH, BSB

23 BSB, BSS, BLT 23 -

24 - 24 -25 - 25 -

26 HLN 26 -

27 HSC, HLN, HSE, BLB, BLT 27 HSN, HSE, HSC, LLF, BLS, CLB

28 BLT 28 HSN, HSE, HSC, LLH, BLS

29 HLH, BLT, BLB 29 HSE, HSC, BLS, BLG, CLB

30 HLH, LLF, BSS, BLB, BLT, CLB 30 LLH


15/17

Table 4. Summary of pooled data of the average duration of grooming in individual mice.

Mouse no. HG LG CG BG

1 4.29 6.05 0 9.13

2 8.15 5.76 6.25 7.59

mean 6.22 5.905 3.125 8.36

sd 2.729432175 0.205060967 4.419417382 1.088944443

Table 5. Summary of pooled data of the average intervalof grooming in individual mice.


1 7.5 8.77 0 9.33

2 7.68 5.8 6.45 7.07

mean 7.59 7.285 3.225 8.2

sd 0.127279221 2.10010714 4.560838739 1.598061325

Table 6. Summary of pooled data of the average frequency of grooming in individual mice.


1 24 19 0 16

2 26 34 4 29

mean 25 26.5 2 22.5

sd 1.414213562 10.60660172 2.828427125 9.192388155

Table 7. Summary of pooled data of the average frequency of grooming in male mice.


1 1 19 0 102 57 83 13 31

3 50 19 4 33

4 1 19 0 10

mean 27.25 35 4.25 21

sd 30.44530615 32 6.130524719 12.72792206


16/17

Table 8. Summary of pooled data of the average frequency of grooming in female mice.


1 13 16 5 10

2 40 52 6 27

3 43 14 4 26

4 13 16 5 0

mean 27.25 24.5 5 15.75

sd 16.5 18.35755975 0.816496581 13.07351011

Table 9. Summary of results of Kruskall-Wallis computations.

Table no. k value N value Tabulated value H value Decision Conclusion

4 4 8 6.167 3.17 Fail to reject Ho No significant differences






17/17

Date post:	05-Apr-2018
Category:	Documents
Upload:	paulo-miguel-kim
View:	224 times
Download:	0 times

Observation and Measurement of Behavior of Mus Musculus

Documents