1 |Mitchell, J.L.

Jaime Lee Mitchell

Bio-3T Lab Report (2)

Dr. Tahamont

23 April 2010

Relationship of Body Depth and Environmental Ammonia Concentrations

Abstract: Alaskan freshwater sticklebacks are among the youngest species known

to exist, having a short evolutionary time span of 22,000 years. This allows

scientists to easily track and monitor several of their evolutionary changes.

Additionally, many differences have been found between the freshwater stickleback

and marine species from which they have evolved (Pennisi, 2000). Therefore,

observation of changes in freshwater sticklebacks is considered to be a valuable

tool in the study of evolution and natural selection.

An experiment was performed with different populations of Gasterosteus

aculeatus, a species of freshwater stickleback obtained from lakes located in Alaska.

This was done in order to formulate a question based on observable phenotypic

differences in 20 specimens of G. aculeatus from three lake populations. Careful

measurements were taken including the pelvic girdle expression, lateral plates, and

body depth sizes. Upon completion, a note was made of any immediate differences

observed. A further comparison of our data with that of our colleagues and

previously data from research performed by Dr. Travis of Rowan University (Travis,

2008) were made. The purpose of our research was to formulate a question and

develop a hypothesis based on the variations observed between several selections

of populations, in order to gain a better understanding of natural selection and

evolution.
2 |Mitchell, J.L.

Our observations showed that higher ammonia concentrations were found in

the lakes of G. aculeatus possessing larger body depths. Therefore, we

hypothesized that increased ammonia levels result in larger body depths. After

thorough research, I found evidence to support this hypothesis although finding

evidence according to our rationalization was proved difficult. As a result, I had

accepted the alternate hypothesis after finding a relationship between high

tolerance of ammonia and large body size in another species of fish. Therefore, I

rejected the null hypothesis that ammonia concentrations have no effect on body

depth sizes in populations of G. aculeatus.

Introduction: Analysis of results using body depth measurements and ammonia

levels were used to compare three populations of G. aculeatus from lakes Jim, Long,

and Bear Paw (Fig. 1). Ammonia concentrations were highest in Jim Lake and

lowest in Long Lake (Fig. 1). It was found that the largest body depths of G.

aculeatus inhabited lakes with the highest ammonia concentrations. We believed

this indicated the possibility of a relationship between large body depth size and

high ammonia levels. Therefore, a hypothesis was formulated stating that

increased ammonia concentrations result in increased body depths. The hypothesis

was based on the rationalization that increased ammonia concentrations result in a

higher yield of phytoplankton, which would suggest the availability of a considerable

food source. As a result, G. aculeatus inhabiting environments with high ammonia

concentrations would exhibit a larger body depth to that of populations in lakes with

lower ammonia concentrations. After carefully researching the question, we

accepted the null hypothesis stating that, high ammonia concentrations have no

effect on the body depth size found in G. aculeatus.

3 |Mitchell, J.L.

Methods and Materials: The experimental protocol followed the lab design

created by Dr. Mathew Travis (2008), titled Natural Selection and Interpopulation

Variation in Threespine Stickleback. Twenty specimens of G. aculeatus from three

different Alaskan lake populations were examined. The samples were pre-stained

with Alizarin red, which binds to the bone to permit easier observation and

measurement. Three different measurements of the external body were performed

in order to make population comparisons.

The body depth was measured starting from the first dorsal spine and

proximal to the pelvic spine. A dial caliper was used to obtain these measurements.

Next, the lateral plates located on the side of the body and just behind the head

were counted. Lastly, the pelvic girdle expression was observed. The measurement

used to identify the pelvic girdle expression is called the pelvic score. This is a

categorical reference used to measure the size, as well as complexity of the pelvic

girdle (Travis, 2008).

Subsequent to performing the measurements, a compilation of our findings

was prepared that included data collected by our colleagues. A thorough analysis

and comparison of the compiled results was made with the lake data (Bell, 2004)

provided by Dr. Travis. Any patterns observed between lake characteristics and

averaged results of our data were documented. Research was executed using

primary literature sources in order to locate evidence supporting our hypothesis.

 4 |Mitchell, J.L.

Figure 1.

The average body depth

sizes in three populations
of G. aculeatus, in
comparison to the
ammonia concentration
levels of their
environment.

The three lakes examined

are Long Lake, Bear Paw,
and Jim Lake. Jim Lake
displays the largest body
depth size, in addition to
the highest ammonia
level. The lowest
ammonia concentrations
are seen in Long Lake,
which possesses a
population of G. aculeatus
with smaller body depths
in comparison with the
two other lakes examined.
5 |Mitchell, J.L.

Results: The average body depth sizes in three populations of G. aculeatus were

compared to the ammonia concentration levels specific to their environment (Fig.1).

The three lakes examined included Long Lake, Bear Paw Lake, and Jim Lake. Jim

Lake displays the largest body depth size of 10.33(mm), in addition to the highest

ammonia level of 20.9 (ppm). The lowest ammonia concentration being 4.0 (ppm)

was found in Long Lake, which possess a population of G. aculeatus with the

smallest body depths averaging 7.59 (mm) in comparison with the two other lakes

examined. Bear Paw displayed intermediate results in both average body depth
6 |Mitchell, J.L.

and ammonia concentration, however, was similar to results found in Long Lake.

The average body depth of Bear Paw was 7.89(mm), while the average ammonia

concentration was at 6.1(mm).

Discussion: The objective of our research was to determine if high ammonia

concentrations contributed to the evolution of G. aculeatus populations possessing

larger body depths. After looking into the diet of G. aculeatus, we found that

several species of phytoplankton as well as zooplankton are considered to be part of

the freshwater stickleback diet (Seehausen, 2009). Interestingly, high ammonia

concentrations can result in an increased biomass in several species of

phytoplankton and zooplankton according to Romo and Vellena (2005). In addition,

research done on juvenile three-spined sticklebacks regarding their choices in prey

showed that biomass of plankton was the determinant in preference. With few

exceptions, plankton species with the largest biomasses had the highest

consumption rates by the juvenile stickleback (Hangelin and Ilppo, 1988).

Additional research showed that limnetic stickleback mainly feed on

phytoplankton, while benthics largely consume zooplankton. Benthic freshwater

stickleback are found to be significantly larger in size than limnetic feeders (Nagel

and Schluter, 1998). Stickleback from Jim Lake, are perhaps, benthic feeders, which

could explain the possession of a larger body depth as compared to Bear Paw and

Long Lake. In addition, benthic fish are often found in bodies of water with low

elevations, such as Jim Lake. However, this is not enough evidence to contribute

high ammonia concentration to the large body depth sizes observed in Jim Lake.

More research and information would be needed to determine whether the

stickleback populations of these lakes are in fact benthic or limnetic feeders.

7 |Mitchell, J.L.

Research done regarding ammonia toxicity and its effect on the growth of the

South African abalone Haliotis midae Linnaeus, showed that tolerance of high

ammonia environments is directly contributed to a larger body size (Reddy-Lopata,

2006). This may provide evidence that natural selection occurred in the high

ammonia environment of Jim Lake. The stickleback fish with larger body depths

may have been favored due to a higher tolerance, and therefore, had better

chances of survival in high ammonia environments. Therefore, I accepted our

alternate hypothesis that high ammonia concentrations result in larger body depths

observed in G. aculeatus. However, more research is required in order to support

the original rationalization that an abundant food source contributes to the

evolution of larger body depths.

Acknowledgments: I would like to thank my Biology 3T lab partners for their

contributions and academic input regarding the development of a hypothesis, in

addition to their assistance with experimental research. I would like to express

appreciation towards my colleagues at Rowan University for any suggestions that

aided my lab partner, Michelle Moormann and I in our investigation and lab report

construction.
8 |Mitchell, J.L.

Works Cited

Bell, M. 1993. Stickleback Lake Information. unpublished data, compliments of Dr.

Matt Travis, personal communication. October 2008. Rowan University.

Hangelin, C. Iippo, V. 1988. Food selection in juvenile three-spined sticklebacks

studied in relation to size, abundance and biomass of prey. Hydrobiologia.

157: 169-177.

Nagel,L., Schluter, D. 1998. Body size, natural selection, and speciation in

sticklebacks. Evolution. 52: 209-218.

Pennisi, E. 2000. Nature steers a predictable course. Science. 287: 207–209.

Reddy-Lopata, K., Averswald, L., Cook, P. 2006. Ammonia toxicity and its effects on

growth of South Africa abalone Halioitis midae Linnaeus. Aquaculture. 261:

678-687.

Romo, S., Villena, M.J. 2003. Phytoplankton strategies and diversity under different

nutrient levels and planktivorous fish densities in a shallow Mediterranean

lake. J. Plankton. Res. 27: 1273-1286.

Seehausen, O. 2009. Speciation affects ecosystems: evidence that speciation and

adaptive radiation can change the properties of an ecosystem is the reminder

9 |Mitchell, J.L.

of the pressing need to integrate ecosystems science and evolutionary

biology. Nature. 458: 1122-1123.

Bibliography

Eppley, R. W., Rogers, J. N., McCarthy, J. J. 1969. Half saturation constants for

uptake of nitrate and ammonium by marine phytoplankton. Limnol.

Oceanogr. 14: 912-920.

Mitchell 2010. Ammonia concentrations vs. body depth in G. aculeatus. Biological

Sciences, Rowan University. pp 20.

Person-Le Ruyet, J., Chartois, H., Quemener, L. 1995. Comparative acute ammonia

toxicity in marine fish and plasma ammonia response. Aquaculture. 136:

181-194.