Laboratories in introductory biology must engage and excite students in ways that effectively prepare them to succeed in upper-level courses in a field with a rapidly increasing body of knowledge. Our inquiry-based lab immerses students in the process of science by asking them to develop behavioral bioassays to test the response of aquatic macroinvertebrates to various pollutants. Students begin with literature review and hypothesis generation on an open-ended question; continue through experimental design, data collection, and analysis; and finish with writing a paper that is peer reviewed. A series of weekly lab activities serves to scaffold key skills that enable completion of the behavioral bioassays, and a final field aquatic-biomonitoring project connects lab work to real-life environmental issues. Because not much is known about how certain pollutants affect the behavior of specific aquatic invertebrates, students’ curiosity is piqued by new discovery, and instructors are engaged by the sense of partnering with students to explore the unknown. This module requires that students engage in core biological concepts, including the significance of variation in living organisms, the structure and function of organisms, and impacts of environmental change on both homeostasis and populations.

Recent educational reform in the STEM disciplines in the United States recommends pedagogical initiatives to increase student engagement in the face of an ever increasing body of knowledge (AAAS, 2011; Association of American Universities, 2011). In the life sciences, a minimal number of core disciplinary foci have been identified, with the recommendation that other content be reduced in order to increase depth of coverage and time for student-centered inquiry-based activities (AAAS, 2011; NGSS Lead States, 2013). Implementation of these reforms will result in an increasing need for activities that are both inquiry-based and connect to core concepts at the introductory level in order to attain the desired depth. We have developed an inquiry-based laboratory curriculum that stimulates active engagement with the process of science while connecting to content in all five core knowledge concepts and five of the six core competencies identified in Vision and Change in Undergraduate Biology Education (AAAS, 2011).

The Laboratory Module

This course module asks students to choose a common freshwater macroinvertebrate and attempt to develop a behavioral bioassay to detect a specific water pollutant in the range of levels typically found in local streams and lakes. The goals of our module were threefold: (1) to engage students in the process of science, (2) to connect real-life environmental issues with classroom content, and (3) to immerse students in inquiry as a way to help them internalize basic biological concepts. The module is spread out over 12 weeks of a typical 15-week semester of introductory biology for undergraduate majors (Table 1). The series of weekly lab exercises scaffolds the skills needed by students to successfully complete their own scientific experiments. The first lab exercise reviews the scientific method as well as how scientific discoveries are communicated and built upon (week 1). Skills for locating high-quality information about a new topic prepare students for later requirements to mine the literature for background material on their experimental organisms and pollutants (week 2). Lab sessions on sources and consequences of variation, as well as animal behavior, hone observational skills and connect to core concepts in the course (evolution/adaptation, structure/function) (weeks 3 and 4). Skills for experimental design, data analysis, and scientific writing are all introduced through the lens of a multi-week bioassay experiment in which students work in teams of two and have the freedom to select their test organism and pollutant for the development of a behavioral bioassay (weeks 5–11). During this time, students plan their experiments using a leading worksheet (Figure 1), prepare a formal proposal, collect and analyze data, write a draft research manuscript, do peer reviews (Figure 2), and prepare a final manuscript (for sample grading rubrics, see Table 2). The module wraps up with a field trip to a local park with headwater streams in the Lake Michigan watershed. Here, we connect the lab portion of the module to real-life water-quality issues with an aquatic biomonitoring activity (week 12). By the end of the module, most students have experienced the excitement of open-ended scientific discovery as well as the challenges of unanticipated outcomes.

Table 1.
Multi-week inquiry-based laboratory accomplishes multiple learning goals.
WeekLab ActivityLearning Objectives: Students Can…AAAS Concept or Competency Addressed
1 Asking questions using termite trail-following behavior (Johnson, 2009
  • Identify questions answerable with the scientific method.

  • Create a testable scientific hypothesis.

 
Competency 1 
2 Communicating scientific discoveries (library field trip) 
  • Locate high-quality relevant literature sources for background information, using both primary and secondary sources.

  • Understand how scientific knowledge is generated and connected.

 
Competencies 1, 5 
3 Measuring characteristics of Arabidopsis thaliana within and between genetic clones 
  • Measure variation in living organisms and connect to possible underlying sources of variation from genes and environment.

 
Concept 1
Competency 1 
4 Creating partial ethograms and then quantifying the behavior of common freshwater invertebrates 
  • Identify quantifiable behaviors in different invertebrate organisms.

  • Connect common environmental pollutants to potential stress for aquatic organisms.

 
Concept 1, 2
Competency 1 
5 Choosing a pollutant and a species for the behavioral bioassay after practicing behavioral observations on all macroinvertebrates available 
  • Explain the value of behavioral bioassays as a method for environmental testing.

  • Find relevant information in published literature that can inform choices for planning a behavior bioassay.

 
Competencies 1, 6 
6 Employing inferential statistical analysis of data and experimental design 
  • Summarize raw data using descriptive statistics and figures.

  • Design an experiment appropriate for a specific inferential test.

 
Competencies 1, 2 
7 Practicing rhetorical writing through proposal development 
  • Write a case for proposed behavioral bioassay with background literature citations and detail about planned methods.

 
Concepts 3, 4
Competencies 1, 5 
8-10 Executing data collection for bioassays under controlled experimental conditions 
  • Collect experimental data under appropriately controlled conditions.

 
Concepts 2, 3, 4
Competencies 1, 4 
11 Completing data analysis and writing research manuscript for peer review 
  • Interpret quantitative data using statistics.

  • Communicate results using figures and scientific writing.

  • Critique the writing of peers and receive critique.

  • Rewrite improved research manuscripts.

 
Concepts 3, 4, 5
Competencies 1, 2, 5 
12 Connecting lab experiment to civic engagement through biomonitoring of a local stream (field trip to park) 
  • Apply lab-based knowledge to field monitoring of the environmental health of a local stream.

 
Concept 5
Competencies 1, 4, 6 
WeekLab ActivityLearning Objectives: Students Can…AAAS Concept or Competency Addressed
1 Asking questions using termite trail-following behavior (Johnson, 2009
  • Identify questions answerable with the scientific method.

  • Create a testable scientific hypothesis.

 
Competency 1 
2 Communicating scientific discoveries (library field trip) 
  • Locate high-quality relevant literature sources for background information, using both primary and secondary sources.

  • Understand how scientific knowledge is generated and connected.

 
Competencies 1, 5 
3 Measuring characteristics of Arabidopsis thaliana within and between genetic clones 
  • Measure variation in living organisms and connect to possible underlying sources of variation from genes and environment.

 
Concept 1
Competency 1 
4 Creating partial ethograms and then quantifying the behavior of common freshwater invertebrates 
  • Identify quantifiable behaviors in different invertebrate organisms.

  • Connect common environmental pollutants to potential stress for aquatic organisms.

 
Concept 1, 2
Competency 1 
5 Choosing a pollutant and a species for the behavioral bioassay after practicing behavioral observations on all macroinvertebrates available 
  • Explain the value of behavioral bioassays as a method for environmental testing.

  • Find relevant information in published literature that can inform choices for planning a behavior bioassay.

 
Competencies 1, 6 
6 Employing inferential statistical analysis of data and experimental design 
  • Summarize raw data using descriptive statistics and figures.

  • Design an experiment appropriate for a specific inferential test.

 
Competencies 1, 2 
7 Practicing rhetorical writing through proposal development 
  • Write a case for proposed behavioral bioassay with background literature citations and detail about planned methods.

 
Concepts 3, 4
Competencies 1, 5 
8-10 Executing data collection for bioassays under controlled experimental conditions 
  • Collect experimental data under appropriately controlled conditions.

 
Concepts 2, 3, 4
Competencies 1, 4 
11 Completing data analysis and writing research manuscript for peer review 
  • Interpret quantitative data using statistics.

  • Communicate results using figures and scientific writing.

  • Critique the writing of peers and receive critique.

  • Rewrite improved research manuscripts.

 
Concepts 3, 4, 5
Competencies 1, 2, 5 
12 Connecting lab experiment to civic engagement through biomonitoring of a local stream (field trip to park) 
  • Apply lab-based knowledge to field monitoring of the environmental health of a local stream.

 
Concept 5
Competencies 1, 4, 6 
Figure 1.

Leading worksheet used to help students plan their behavioral bioassay projects.

Figure 1.

Leading worksheet used to help students plan their behavioral bioassay projects.

Figure 2.

Assignment sheet for peer review of colleagues’ research manuscripts. Readings are from McMillan (2012).

Figure 2.

Assignment sheet for peer review of colleagues’ research manuscripts. Readings are from McMillan (2012).

Table 2.
Grading rubrics showing expectations for exemplary research proposals and research manuscripts for behavioral bioassay projects.
Research ProposalResearch Manuscript
SectionExpectationsSectionExpectations
Cover Sheet Includes names, institution, majors, contact information, project title, and date Cover page Includes title, authors’ names, affiliation; title includes scientific name of organism and is neither too vague nor too detailed 
Purpose of Project Provides background research about both organism and stressor; describes why each is appropriate for this research and connects all into a coherent whole; gives a clear hypothesis for the study Abstract Complete, concise summary of entire paper; flows easily; includes relevant statistical result in format (T =, N =, P =) 
Significance Clearly places work in larger context, gives relevance to planned research Introduction Provides rationale for conducting experiment, including background and relevance of organism and stressor; describes relevance of conducting a behavioral bioassay; ends with identification of hypothesis being tested and brief summary of how it will be tested. Flows logically. 
Plan of Work Presents all steps and is logical and adequately detailed with experimental design and sample size planned; includes statistical test to be used; sampling is appropriate to the problem Materials & Methods Must be adequately detailed; identifies descriptive and inferential statistics used; all written in past tense; flows logically 
Anticipated Result Description shows a clear understanding of whole project Results Quantitative results summarized well in text and correctly labeled figure or table; mean, standard deviation, and range included. Qualitative results reported. Reader referred to figures/tables. No raw data. 
Supplies Complete list of everything needed as described in Materials & Methods Discussion Discusses, interprets, and explains meaning of quantitative and qualitative results. Places interpretation in larger context introduced in Introduction. Considers alternative hypotheses for results. Cites appropriate literature. Flows logically from start to finish. 
Literature Cited At least two scholarly sources in CSE author/year format, both here and in text of document. Acknowledgments At least one acknowledgement included; written in complete sentence(s). 
  Literature Cited At least three scholarly sources in proper
CSE format; correct in-text format. 
  Appendix Includes raw data, H0 and Ha, statistical calculations (Excel or handwritten) 
Research ProposalResearch Manuscript
SectionExpectationsSectionExpectations
Cover Sheet Includes names, institution, majors, contact information, project title, and date Cover page Includes title, authors’ names, affiliation; title includes scientific name of organism and is neither too vague nor too detailed 
Purpose of Project Provides background research about both organism and stressor; describes why each is appropriate for this research and connects all into a coherent whole; gives a clear hypothesis for the study Abstract Complete, concise summary of entire paper; flows easily; includes relevant statistical result in format (T =, N =, P =) 
Significance Clearly places work in larger context, gives relevance to planned research Introduction Provides rationale for conducting experiment, including background and relevance of organism and stressor; describes relevance of conducting a behavioral bioassay; ends with identification of hypothesis being tested and brief summary of how it will be tested. Flows logically. 
Plan of Work Presents all steps and is logical and adequately detailed with experimental design and sample size planned; includes statistical test to be used; sampling is appropriate to the problem Materials & Methods Must be adequately detailed; identifies descriptive and inferential statistics used; all written in past tense; flows logically 
Anticipated Result Description shows a clear understanding of whole project Results Quantitative results summarized well in text and correctly labeled figure or table; mean, standard deviation, and range included. Qualitative results reported. Reader referred to figures/tables. No raw data. 
Supplies Complete list of everything needed as described in Materials & Methods Discussion Discusses, interprets, and explains meaning of quantitative and qualitative results. Places interpretation in larger context introduced in Introduction. Considers alternative hypotheses for results. Cites appropriate literature. Flows logically from start to finish. 
Literature Cited At least two scholarly sources in CSE author/year format, both here and in text of document. Acknowledgments At least one acknowledgement included; written in complete sentence(s). 
  Literature Cited At least three scholarly sources in proper
CSE format; correct in-text format. 
  Appendix Includes raw data, H0 and Ha, statistical calculations (Excel or handwritten) 

Behavioral bioassays are used around the world to monitor freshwater resources (Bae & Park, 2014). In this module, students can see the potential practical application of their research. When altered behavior is used in an assay to test for pollution instead of death, the assay can be much more sensitive to even very small changes in water chemistry (Alonso et al., 2009) and can be an excellent way to detect the early signs of water-quality problems (Mills et al., 2006; Melvin & Wilson, 2013). These biological early-warning systems can be limited by our knowledge of how water pollution will affect the behavior of aquatic animals (Gerhardt et al., 2006). Thus, student research projects that explore how animals react to specific pollutants can be connected to real environmental problems and solutions. For example, one group counted the number of times that 20 different planaria contracted and then elongated in a 3-minute period when each worm was placed first in control spring water and then, after a 3-minute break, in the treatment of spring water with caffeine (2 µL). They found that the worms increased their contraction rate by 6× when in the caffeine treatment. Thus, they had developed a behavior bioassay using planaria that could be used to detect the presence of a contaminant in waste water by simply watching the worms move around their environment.

We use six different aquatic macroinvertebrates for these experiments because that gives students an adequate array of choices to ensure a large variety of projects within a lab section. The species we offer are easily obtained at low cost (i.e., local pet store, Carolina Biological Supply), making adequate sample sizes (10–20 per team) possible in student experiments (Table 3). In addition, they are very observable in a lab situation and, from what is known of their behavior and physiology, they are likely to show sensitivity to various common water pollutants. Each semester, we try to offer at least five stressor pollutants for experimentation (Table 4). We attempt to offer ones with minimal safety concerns, both for the students and the environment (through disposal of waste solutions). We also try to choose pollutants that are common in our local waterways, to help students connect to the environment around them. Recent interest in pharmaceuticals as pollutants in freshwater systems (Stackelberg et al., 2004) has inspired us to try offering some of these as choices for the bioassays.

Table 3.
Easily acquired freshwater macroinvertebrates provided for students for behavioral bioassay development.
OrganismReferences for Behavioral ResponsesBasic Supplies Needed (All Animals Kept in Spring Water)
Water Slater (Asellus sp.) Bundschuh et al., 2012  2-L round glass holding containers, spoons for animal transfer, finger bowls for observing individuals 
Water Flea (Daphnia sp.) Gerhardt et al., 2005  1-L beaker for holding, plastic pipette for animal transfer, watch glass or deep-well slides and dissecting microscope for making observations 
Planaria (Dugesia sp.) Pagan et al., 2009  Finger bowls for holding, plastic pipette for animal transfer, watch glass and dissecting microscope for making observations 
Scud (Gammarus sp.) Mills et al., 2006  2-L round glass holding containers, spoons for animal transfer, finger bowls for observing individuals 
Aquatic Worm (Lumbricus sp.) West & Ankley, 1998  Finger bowls for holding, pipettes for transfer, watch glasses for observation 
Ghost Shrimp (Paleomonetes sp.) Farr, 1977  Aquarium with airstone for holding, nets for animal transfer, large finger bowls with lids for observations 
OrganismReferences for Behavioral ResponsesBasic Supplies Needed (All Animals Kept in Spring Water)
Water Slater (Asellus sp.) Bundschuh et al., 2012  2-L round glass holding containers, spoons for animal transfer, finger bowls for observing individuals 
Water Flea (Daphnia sp.) Gerhardt et al., 2005  1-L beaker for holding, plastic pipette for animal transfer, watch glass or deep-well slides and dissecting microscope for making observations 
Planaria (Dugesia sp.) Pagan et al., 2009  Finger bowls for holding, plastic pipette for animal transfer, watch glass and dissecting microscope for making observations 
Scud (Gammarus sp.) Mills et al., 2006  2-L round glass holding containers, spoons for animal transfer, finger bowls for observing individuals 
Aquatic Worm (Lumbricus sp.) West & Ankley, 1998  Finger bowls for holding, pipettes for transfer, watch glasses for observation 
Ghost Shrimp (Paleomonetes sp.) Farr, 1977  Aquarium with airstone for holding, nets for animal transfer, large finger bowls with lids for observations 
Table 4.
Representative pollutant stressors available for behavioral bioassays.
Pollutant StressorReference for Environmental LevelsEquipment Needed
Caffeine Stackelberg et al., 2004  Premixed stock solution, micropipettes 
Fertilizer Mason et al., 1990  Granular ammonium nitrate, digital balance 
Glyphosate Goldsborough & Beck, 1989  Roundup® herbicide, micropipettes 
pH (acidity) Havens et al., 1993  pH buffer capsules from Micro Essential Laboratory, pH meter 
Salt Williams et al., 1999  Table salt or sidewalk salt crystals, digital balance 
Pollutant StressorReference for Environmental LevelsEquipment Needed
Caffeine Stackelberg et al., 2004  Premixed stock solution, micropipettes 
Fertilizer Mason et al., 1990  Granular ammonium nitrate, digital balance 
Glyphosate Goldsborough & Beck, 1989  Roundup® herbicide, micropipettes 
pH (acidity) Havens et al., 1993  pH buffer capsules from Micro Essential Laboratory, pH meter 
Salt Williams et al., 1999  Table salt or sidewalk salt crystals, digital balance 

At the end of this lab module, we partner with a local land trust to help monitor the health of headwater streams in the local watershed. Students are assigned background reading material about pollution in watersheds and the use of aquatic biomonitoring to detect impairments to water quality. Then, during a field trip, each lab group of 12–24 students is given a specific reach of a headwater stream to sample with dip nets for aquatic macroinvertebrates. Students capture wild organisms from the streams, identify them using simple keys, and place them in pollution tolerance categories. We use a citizen science protocol developed by Hoosier Riverwatch (2015) to calculate a pollution tolerance index for each sampling site and discuss with students possible reasons for any impairments detected in the watershed. We provide all our data to the land trust for analysis of long-term changes in these streams as a result of their ecological restoration of the surrounding landscape. Because many of the macroinvertebrates we have been using in the lab experiments have closely related species present in local waterways, students can make the connection between their behavioral bioassays and the health of local ecosystems. Our university is located in the Lake Michigan watershed, and we further connect the health of headwater streams to the quality of regional resources – in our case swimming beaches, but in other areas this could be fishing areas or drinking water.

Results

In the past 6 years that we have run this lab module, students have shown a strong preference to develop bioassays for ghost shrimp (Paleomonetes sp.) and pH, choosing these topics in 32 and 28 of a total of 131 studies, respectively – perhaps because of familiarity with the aquarium trade and acid rain from popular media. Aquatic worms (Lumbricus variegatus) and glyphosate were not popular, chosen <10% of the time, and a few organism–stressor combinations have never been chosen (Table 5). A recent addition of caffeine as a pharmaceutical pollutant available for testing proved extremely popular; half of the teams from all laboratory sections chose this as their stressor in the semester it was introduced. Students used a wide variety of quantifiable behaviors for their experiments, mostly related to feeding or predator avoidance. Although we directed them to consider real environmental levels of pollutants in their bioassays (Table 4), we also found them choosing a wide range of pollutant values. Teams were encouraged to have only two treatments in their experiments, a control and one experimental level of the stressor pollutant for simplicity of statistical analysis. The behavioral-bioassay combinations most likely to result in significant differences in behavior appeared to be planaria/salt and scuds/pH, but many combinations have not been tried more than a couple times in our course. This, along with a wide range of experimental designs, makes it difficult to say which animals and pollutants are most likely to result in significant changes in behavior.

Table 5.
Results of student-designed behavioral bioassays over 6 years of running the course module: ratios of experiments that resulted in significant differences between stressor treatment and control to total number of experiments run. Ranges of stressor levels that resulted in a significant response are listed.
Species with Example BehaviorsCaffeine (0.0016–500 ug/L)Fertilizer (0.4–12 mg/L)Glyphosate (105 ug/L)Salt (0.2–6000 mg/L)pH (3.98–6.0)Totals
Asellus sp. Crawling rate Antenna wipes 1:2 0:0 0:1 2:3 7:10 10:16 
Daphnia sp. Spinning rate Wall contact 1:3 0:1 0:2 5:8 5:11 11:25 
Dugesia sp. Time spent contracted Flip rate 2:3 4:7 0:1 5:6 2:4 13:21 
Gammarus sp. Tail flicking rate Time spent moving 1:3 1:1 0:0 4:13 7:9 13:26 
Lumbricus sp.
Body crossing rate Time spent clumped 
0:1 2:4 1:1 0:1 2:4 5:11 
Paleomonetes sp. Forward swimming time Pincher extension rate 5:13 3:6 0:2 1:2 5:9 14:32 
Totals 10:25 10:19 1:7 17:33 28:47 66:131 
Species with Example BehaviorsCaffeine (0.0016–500 ug/L)Fertilizer (0.4–12 mg/L)Glyphosate (105 ug/L)Salt (0.2–6000 mg/L)pH (3.98–6.0)Totals
Asellus sp. Crawling rate Antenna wipes 1:2 0:0 0:1 2:3 7:10 10:16 
Daphnia sp. Spinning rate Wall contact 1:3 0:1 0:2 5:8 5:11 11:25 
Dugesia sp. Time spent contracted Flip rate 2:3 4:7 0:1 5:6 2:4 13:21 
Gammarus sp. Tail flicking rate Time spent moving 1:3 1:1 0:0 4:13 7:9 13:26 
Lumbricus sp.
Body crossing rate Time spent clumped 
0:1 2:4 1:1 0:1 2:4 5:11 
Paleomonetes sp. Forward swimming time Pincher extension rate 5:13 3:6 0:2 1:2 5:9 14:32 
Totals 10:25 10:19 1:7 17:33 28:47 66:131 

As part of a standard course evaluation, students were asked what they liked most about the entire course curriculum, and many identified this lab module as their favorite experience. Many reported that the bioassay experiment gave them more confidence overall in writing and learning about science. Several students talked about how the course had changed their attitudes or opened their eyes to their connection with the ecosystems around them. On a student assessment of learning gains administered at the end of the course in 2013 (and filed at http://salgsite.org), 87% reported at least moderate gains in using a critical approach to analyzing data and arguments in their daily life. Even more encouraging, 98% of the responding students said that they gained a better understanding of the scope and complexity of solutions to Great Lakes environmental challenges as a result of this course. Many students commented that the field trip served as a capstone experience, helping them make connections between the course content, laboratory research, and their own immediate surroundings.

Uses of the Module

This lab module was designed for beginning undergraduate biology majors but could be scaled up or down for different levels of students. The experiments would be simple enough for nonmajor environmental students, especially if pollution treatment levels were proscribed and appropriate solutions provided (rather than having students create their own). Selected single experiments from this module could be developed for use at the K–12 level. For example, a 1-day lab exercise on the effect of salt contamination on water slaters’ crawling rate would make an excellent and inexpensive lab for middle or high school students and could be tied in nicely to impacts of winter road salt on aquatic ecosystems. In addition, upper-level undergraduates could be asked to explore the mechanisms behind the behavioral changes observed during the bioassay experiments, connecting to core concepts in physiology. We have found that these relatively simple experiments provide excellent practice for communicating about research. At this early undergraduate level, we devote a significant portion of time in this module to developing scientific writing skills, including writing proposals, reviewing published literature, preparing research manuscripts, and peer review.

Conclusions

This lab module on developing behavioral bioassays for aquatic organisms engages students in the excitement of discovery while connecting the science to real-life problems in our region. It introduces core concepts of how natural variation is key in the process of evolution leading to adaptation, how structure (especially behavior) of an organism is tied to functions for feeding or escape, and how organisms interact with a changing environment. Students leave this introductory course with a set of basic skills such as literature search, experimental design, and scientific writing that will be used again and again in our upper-level biology courses and beyond. Most importantly, students have witnessed that biology is not a science of memorizing facts and getting “correct” answers in lab exercises, but one of curiosity about the living world, fostering enthusiasm to learn more about it.

We thank undergraduates at Valparaiso University, both those enrolled in the introductory course and those working as laboratory aides, who helped us fully develop this module. Special thanks to Paul Quinlan and other staff at the Shirley Heinz Land Trust in Valparaiso, IN, and Laura Barghusen of Openlands in Chicago, IL, for their work with our students in monitoring headwater steams.

References

References
AAAS
(
2011
).
Vision and Change in Undergraduate Biology Education: A Call to Action
. Available online at http://visionandchange.org/.
Alonso, A., De Lange, H.J. & Peeters, E.T.H.M. (
2009
).
Development of a feeding behavioural bioassay using the freshwater amphipod Gammarus pulex and the multispecies freshwater biomonitor
.
Chemosphere
,
75
,
341
346
.
Association of American Universities
(
2011
).
Framework for systemic change in undergraduate STEM teaching and learning
.
Washington, DC
:
Association of American Universities
.
Bae, M.-J. & Park, Y.-S. (
2014
).
Biological early warning system based on the responses of aquatic organisms to disturbances: a review
.
Science of the Total Environment
,
466–467
,
635
649
.
Bundschuh, M., Appeltauer, A., Dabrunz, A. & Schulz, R. (
2012
).
Combined effect of invertebrate predation and sublethal pesticide exposure on the behavior and survival of Asellus aquaticus (Crustacea; Isopoda)
.
Archives in Environmental Contamination and Toxicology
,
63
,
77
85
.
Farr, J.A. (
1977
).
Impairment of antipredator behavior in Palaemonetes pugio by exposure to sublethal doses of parathion
.
Transactions of the American Fisheries Society
,
106
,
287
290
.
Gerhardt, A., Ingram, M.K., Kang, I.J. & Ulitzur, S. (
2006
).
In situ on-line toxicity biomonitoring in water: recent developments
.
Environmental Toxicology and Chemistry
,
25
,
2263
2271
.
Gerhardt, A., Janssens de Bisthoven, L. & Soares, A.M.V. (
2005
).
Evidence for the stepwise stress model: Gambusia holbrooki and Daphnia magna under acid mine drainage and acidified reference water stress
.
Environmental Science and Technology
,
39
,
4150
4158
.
Goldsborough, L.G. & Beck, A.E. (
1989
).
Rapid dissipation of glyphosate in small forest ponds
.
Archives of Environmental Contamination and Toxicology
,
18
,
537
544
.
Havens, K.E., Yan, N.D. & Keller, W. (
1993
).
Lake acidification: effects on crustacean zooplankton populations
.
Environmental Science and Technology
,
27
,
1621
1624
.
Hoosier Riverwatch
(
2015
).
Volunteer stream monitoring training manual
. Available online at http://www.in.gov/idem/riverwatch/files/volunteer_monitoring_manual.pdf.
Johnson, A.D. (
2009
).
40 Inquiry Exercises for the College Biology Lab
.
Arlington, VA
:
NSTA Press
.
Mason, J.W., Wegner, G.D., Quinn, G.I. & Lange, E.L. (
1990
).
Nutrient loss via groundwater discharge from small watersheds in southwestern and south central Wisconsin
.
Journal of Soil and Water Conservation
,
45
,
327
331
.
McMillan, V.E. (
2012
).
Writing Papers in the Biological Sciences
, 5th Ed.
Boston, MA
:
Bedford/St. Martin's
.
Melvin, S.D. & Wilson, S.P. (
2013
).
The utility of behavioral studies for aquatic toxicology testing: a meta-analysis
.
Chemosphere
,
93
,
2217
2223
.
Mills, C.L., Shukla, D.H. & Compton, G.J. (
2006
).
Development of a new low cost high sensitivity system for behavioural ecotoxicity testing
.
Aquatic Toxicology
,
77
,
197
201
.
NGSS Lead States
(
2013
).
Next Generation Science Standards: For States, By States
.
Washington, DC
:
National Academies Press
.
Pagan, O.R., Coudron, T. & Kaneria, T. (
2009
).
The flatworm planaria as a toxicology and behavioral pharmacology animal model in undergraduate research experiences
.
Journal of Undergraduate Neuroscience Education
,
7
,
A48
A52
.
Stackelberg, P.E., Furlong, E.T., Meyer, M.T., Zaugg, S.D., Henderson, A.K. & Reissman, D.B. (
2004
).
Persistence of pharmaceutical compounds and other organic wastewater contaminants in a conventional drinking-water-treatment plant
.
Science of the Total Environment
,
329
,
99
113
.
West, C.W. & Ankley, G.T. (
1998
).
A laboratory assay to assess avoidance of contaminated sediments by the freshwater oligochaete Lumbriculus variegatus
.
Archives of Environmental Contamination and Toxicology
,
35
,
20
24
.
Williams, D.D., Williams, N.E. & Cao, Y. (
1999
).
Road salt contamination of groundwater in a major metropolitan area and development of a biological index to monitor its impact
.
Water Research
,
34
,
127
138
.