DIVERSITY OF MACROINVERTEBRATES IN TRIBUTARIES OF THE JACKS

FORK AND CURRENT RIVERS, OZARK NATIONAL SCENIC RIVERWAYS,

MISSOURI AND EFFICACY OF SPRING-FED TRIBUTARIES AS REFUGIA

______

A Dissertation

presented to

the Faculty of the Graduate School

at the University of Missouri-Columbia

______

In Partial Fulfillment

of the Requirements for the Degree

Doctor of Philosophy

______

by

RACHEL L. S. HETH

Robert W. Sites, Dissertation Supervisor

MAY 2015

The undersigned, appointed by the dean of the Graduate School, have examined the dissertation entitled

DIVERSITY OF MACROINVERTEBRATES IN TRIBUTARIES OF THE JACKS

FORK AND CURRENT RIVERS, OZARK NATIONAL SCENIC RIVERWAYS,

MISSOURI AND EFFICACY OF SPRING-FED TRIBUTARIES AS REFUGIA

presented by Rachel L. S. Heth a candidate for the degree of doctor of philosophy, and hereby certify that, in their opinion, it is worthy of acceptance.

Robert W. Sites, PhD, Dissertation Supervisor, Division of Plant Sciences

Deborah L. Finke, PhD, Division of Plant Sciences

Richard M. Houseman, PhD, Division of Plant Sciences

Barry C. Poulton, PhD, United States Geological Survey

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ACKNOWLEDGEMENTS

I thank my committee members for their feedback and support: Robert Sites,

Deborah Finke, Barry Poulton, and Richard Houseman. Financial assistance was provided by the Life Sciences Fellowship and the Gus T. Ridgel Fellowship. Needed field equipment was borrowed from Craig Paukert’s laboratory in the University of

Missouri Fisheries and Wildlife Department and from the Missouri Southern State

University Biology Department. I thank Kristopher Corbett, Justin Doherty, Jack Grant,

Robert Heth, Jordan Holtzwarth, John Layng, Grace Lindner, James Pflug, Kris Simpson,

Robert Sites, Daniel Reynoso-Velasco, and Jessica Warwick for their help and wonderful work ethic in both the field and laboratory. Brandy Bergthold from the Missouri

Department of Natural Resources was invaluable in helping me learn to identify the

Chironomidae. Samplers were designed and made due to the tireless efforts of my parents, Robert (construction) and Ollie Heth (sewing), and to our entomology museum curator Kris Simpson (sewing) to whom I am eternally grateful. Field work in Ozark

National Scenic Riverways was possible because of permission granted by the National

Park Service and Missouri Department of Conservation. I also am grateful for the love, prayers, encouragement, and inspiration I receive from my amazing family: Gabrielle,

Ollie, and Robert Heth.

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TABLES

1. Environmental measures collected in July 2011 before slack-Surber samples were collected from tributaries of the Current River, Missouri. T = Tributary, UCR = upstream of tributary in Current River, DCR = downstream of tributary in Current River. Values are based on means from three subsamples ...... 43

2. Environmental measures collected in January 2012 before slack-Surber samples were collected from tributaries of the Current River, Missouri. T = Tributary, UCR = upstream of tributary in Current River, DCR = downstream of tributary in Current River. Values are based on means from three subsamples ...... 44

3. Two-way analysis of variance tests of the most abundant taxa in July 2011 (> 1% of total sample) between tributaries or among locations in the Current River, Missouri. Dashes indicate data did not meet assumptions of ANOVFA after transformation ....45

4. Two-way analyses of variance of the most abundant taxa in January 2012 (> 1% of total sample) between tributaries or among locations in the Current River, Missouri. Dashes indicate data did not meet assumptions of ANOVA after transformation ...... 47

5. Multiple response permutation tests comparing macroinvertebrate community composition between tributaries and among locations in July 2011 and January 2012 from the Current River, Missouri...... 49

6. Three-way analysis of variance comparing macroinvertebrate movement direction (between drift and upstream movement samplers) ...... 50

7. Two-way analysis of variance tests comparing macroinvertebrate quantities in movement samplers between tributaries and among locations in the Current River, Missouri. Mov. = movement ...... 51

8. Two-way analysis of variance comparing environmental variables using tributary type and location as factors in July 2011 of the Current River, Missouri ...... 52

9. Two-way analysis of variance comparing environmental variables between tributaries or among locations in January 2012 of the Current River, Missouri ...... 53

10. Environmental variables taken in July and September 2013 before Brown vacuum samples were collected in mesohabitats from surface-fed tributaries of the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation, D.O. = Dissolved oxygen, Wentwth. = Wentworth. Values are based on means from three subsamples ...... 108

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11. Environmental variables taken in July and September 2013 before Brown vacuum samples were collected in mesohabitats from spring-fed tributaries of the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation, D. O. = Dissolved oxygen, Wentwth. = Wentworth. Values are based on means from three subsamples. Values are based on means from three subsamples ...... 109

12. Two-way analyses of variance comparing metrics between tributaries and among mesohabitats in July and September 2013 from the Current and Jacks Fork rivers, Missouri ...... 110

13. Jaccard’s similarity scores (J) for pairwise comparisons between tributaries and mesohabitats of the Current and Jacks Fork rivers, Missouri in July and September 2013...... 111

14. Multi-response permutation procedure tests associated with nonmetric multidimensional scaling between tributaries and among mesohabitats in the Current and Jacks Fork rivers, Missouri in July and September 2013. Community refers to macroinvertebrate community. R = riffle, P = pool, V – marginal vegetation. FFG = functional feeding group ...... 112

15. Two-way analyses of variance of functional feeding group (FFG) richness between tributaries and among mesohabitats of the Current and Jacks Fork rivers, Missouri in July and September 2013 ...... 113

16. Two-way analyses of variance of functional feeding group (FFG) densities between tributaries and among mesohabitats of the Current and Jacks Fork rivers, Missouri in July and September 2013 ...... 114

17. Two-way analyses of variance of environmental variables between tributaries and among mesohabitats in the Current and Jacks Fork rivers, Missouri in July and September 2013. Dashes indicate data did not meet assumptions of ANOVA after transformation ...... 115

18. Chironomidae subfamilies and genera collected from Brown vacuum samples in the Current and Jacks Fork river tributaries, Missouri in July and September 2013 ...... 161

19. Three-way analyses of variance testing among metrics or indices for macroinvertebrates collected from the Current or Jacks Fork rivers, Missouri in July 2013. Factors are genera of Chironomidae inclusion, tributary type, and mesohabitat. BI = biotic index, SCI = Stream Condition Index, H’ = Shannon’s diversity index, TR = taxonomic richness, Chiro = genera of Chironomidae inclusion ...... 162

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20. Two-way analysis of variance comparing taxonomic richness of Chironomidae using factors of tributaries and mesohabitats for macroinvertebrates collected from the Current or Jacks Fork rivers, Missouri in July 2013. TR = taxonomic richness ...... 163

21. Multi-response permutation procedure tests associated with nometric multidimensional scaling of macroinvertebrate communities from mesohabitats with genera of Chironomidae excluded or included in the Current and Jacks Fork rivers, Missouri in July 2013. Surface = surface-fed tributaries, Spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation ...... 164

22. Multi-response permutation procedure tests associated with nonmetric multidimensional scaling of communities of Chironomidae in mesohabitats from the Current and Jacks Fork rivers, Missouri in July 2013. Surface = surface-fed tributaries, Spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation ...... 165

23.Two-way analyses of variance comparing subfamily densities of Chironomidae between tributaries and among mesohabitats from the Current and Jacks Fork rivers, Missouri in July and September 2013 ...... 166

24. Multiple linear regressions of environmental variables in relation to densities of the 10 most common genera of Chironomidae in July and September 2013 from the Current and Jacks Fork rivers, Missouri. C/O = Cricotopus/Orthocladius, T = Thienemannimyia. Dashes indicate no variables were statistically significant ...... 167

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LIST OF FIGURES

1. Macroinvertebrate sampling sites along Current River, Missouri. Current River is shown in bold. Big Creek, Sinking Creek, and Rocky Creek are surface-fed tributaries flowing into the Current River. Pulltite Spring, Round Spring, and Blue Spring are spring-fed tributaries flowing into the Current River ...... 54

2. Current River locations within a stream confluence. T = tributary, UCR = upstream of tributary in Current River, DCR = downstream of tributary in Current River. Arrows indicate direction of flow. Circles represent the number of subsamples collected within locations from riffle/run habitat ...... 55

3. Slack-Surber sampler to collect standing macroinvertebrate communities ...... 56

4. Upstream-movement sampler to collect macroinvertebrates crawling upstream ...... 57

5. Drift sampler to collect macroinvertebrates moving downstream ...... 58

6. Most abundant macroinvertebrates collected from the Current River, Missouri from slack-Surber samples collected in July 2011 ...... 59

7. Two-way analysis of variance comparing Amnicola density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Columns represent means of 3 replicates ± standard errors ...... 60

8. Two-way analysis of variance comparing Lepidostoma density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Columns represent means of three replicates ± standard errors ..61

9. Two-way analysis of variance comparing Tricorythodes density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Columns represent means of three replicates ± standard errors. Note the different arrangement of axis labels ...... 62

10. Two-way analysis of variance comparing Cheumatopsyche density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a

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log transformation. Columns represent means of three replicates ± standard errors. Note the different arrangement of axis labels ...... 63

11. Two-way analysis of variance comparing Chironomidae density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Columns represent means of three replicates ± standard errors ..64

12. Most common macroinvertebrates collected from the Current River, Missouri from slack-Surber samples in January 2012 ...... 65

13. Two-way analysis of variance comparing Psychomyia flavida density differences among locations in January 2012 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Columns represent means of three replicates ± standard errors. Note the different arrangement of axis labels ...... 66

14. Two-way analysis of variance comparing Amnicola density differences among locations in January 2012 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Columns represent means of three replicates ± standard errors ..67

15. Nonmetric multidimensional scaling plot of macroinvertebrate communities in July 2011 slack-Surber samples from the Current River, Missouri. Stress = 9.568, randomization p = 0.008 ...... 68

16. Nonmetric multidimensional scaling plot of macroinvertebrate communities in July 2011 slack-Surber samples from the Current River, Missouri. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 9.568, randomization p =0.008 ...... 69

17. Nonmetric multidimensional scaling plot of macroinvertebrate communities in January 2011 slack-Surber samples from the Current River, Missouri. Stress = 7.688, randomization p = 0.004 ...... 70

18. Nonmetric multidimensional scaling plot of macroinvertebrate communities in January 2012 slack-Surber samples from the Current River, Missouri. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River,

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DCR = downstream of the tributary confluence in the Current River, stress = 7.688, randomization p = 0.004 ...... 71

19. Two-way analysis of variance comparing macroinvertebrate drift among locations in January 2012 from the Current River, Missouri. Columns represent means of three replicates ± standard errors ...... 72

20. Nonmetric multidimensional scaling plot of macroinvertebrate communities in July 2011 drift samples from the Current River, Missouri. Stress = 11.698, randomization p = 0.004 ...... 73

21. Nonmetric multidimensional scaling plot for macroinvertebrate communities in July 2011 drift samples from the Current River, Missouri. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 11.698, randomization p = 0.004...... 74

22. Nonmetric multidimensional scaling plot of macroinvertebrate communities in January 2012 drift samples from the Current River, Missouri. Stress = 10.906, randomization p = 0.004 ...... 75

23. Nonmetric multidimensional scaling plot of macroinvertebrate communities in January 2012 drift samples from the Current River, Missouri. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 10.906, randomization p = 0.004 ...... 76

24. Nonmetric multidimensional scaling biplot of macroinvertebrate community correlations with environmental variables in July 2011 slack-Surber samples from the Current River, Missouri. Arrow length indicates strength of correlation. Arrow direction indicates increase environmental values. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 9.568, randomization p = 0.004...... 77

25. Two-way analysis of variance comparing temperature differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Columns represent means of three replicates ± standard errors ...... 78

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26. Two-way analysis of variance comparing dissolved oxygen differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Columns represent means of three replicates ± standard errors ...... 79

27. Two-way analysis of variance comparing embeddedness cover differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Columns represent means of three replicates ± standard errors. Note the different arrangement of axis labels ..80

28. Nonmetric multidimensional scaling biplot of macroinvertebrate community correlations with environmental variables in January 2012 slack-Surber samples from the Current River, Missouri. Arrow length indicates strength of correlation. Arrow direction indicates increase environmental values. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 7.688, randomization p = 0.004...... 81

29. Two-way analysis of variance comparing temperature differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Columns represent means of three replicates ± standard errors ...... 82

30. Two-way analysis of variance comparing substrate size differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Columns represent means of three replicates ± standard errors ...... 83

31. Two-way analysis of variance comparing dissolved oxygen differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Columns represent means of three replicates ± standard errors. Note differences in arrangement of axis labels ...... 84

32. Two-way analysis of variance comparing pH differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of

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tributary confluence in the Current River. Columns represent means of three replicates ± standard errors. Note differences in arrangement of axis labels ...... 85

33. Two-way analysis of variance comparing macrophyte cover differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Columns represent means of three replicates ± standard errors ...... 86

34. Two-way analysis of variance comparing periphyton cover differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Columns represent means of three replicates ± standard errors ...... 87

35. Macroinvertebrate sampling sites in July and September 2013 from surface-fed tributary sites (Ashley Creek, Bay Creek, Big Creek, Rocky Creek, Sinking Creek) and spring-fed tributary sites (Alley Spring, Blue Spring, Cave Spring, Pulltite Spring, Round Spring). Tributaries are associated with the Current and Jacks Fork rivers, Missouri, which are shown in bold ...... 117

36. Brown vacuum sampler used to collect macroinvertebrates in July 2013 from the Current and Jacks Fork rivers, Missouri ...... 118

37. Two-way analysis of variance comparing dominance among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors ...... 119

38. Two-way analysis of variance comparing Shannon’s Diversity Index among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 120

39. Two-way analysis of variance comparing taxonomic richness among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors ...... 121

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40. Composite graph of order-level richness among mesohabitats from July 2013 in the Current and Jacks Fork rivers, Missouri. Surface = surface-fed, Spring = spring-fed, Veg = marginal vegetation ...... 122

41. Nonmetric multidimensional scaling plot of macroinvertebrate communities collected from Brown vacuum samples in July 2013 showing functional feeding group richness from the Current and Jacks Fork rivers, Missouri. Stress = 7.47, randomization p = 0.004...... 123

42. Nonmetric multidimensional scaling plot of macroinvertebrate communities collected from Brown vacuum samples in July 2013 showing functional feeding group richness among mesohabitats from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, Spring = spring-fed tributaries, Veg = marginal vegetation. Stress = 7.47, randomization test p = 0.004 ...... 124

43. Two-way analysis of variance comparing collectors richness among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors ...... 125

44. Two-way analysis of variance comparing filterer richness among mesohabitats in July and September 2013 collected by Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors ...... 126

45. Two-way analysis of variance comparing scraper richness among mesohabitats in July and September 2013 collected by Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors ...... 127

46. Nonmetric multidimensional scaling plot of macroinvertebrate communities collected by Brown vacuum samples in July 2013 showing densities within functional feeding groups from the Current and Jacks Fork rivers, Missouri. Stress = 10.85, randomization p = 0.004 ...... 128

47. Nonmetric multidimensional scaling plot of macroinvertebrate communities collected by Brown vacuum samples in July 2013showing densities within functional feeding groups among mesohabitats from the Current and Jacks Fork rivers, Missouri.

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Surface = surface-fed tributaries, Spring = spring-fed tributaries, Veg = marginal vegetation, Stress = 10.85, randomization p = 0.004 ...... 129

48. Nonmetric multidimensional scaling plot of macroinvertebrate communities collected by Brown vacuum samples in July 2013 showing macroinvertebrate communities from the Current and Jacks Fork rivers, Missouri. Stress = 14.3, randomization p = 0.004 ...... 130

49. Nonmetric multidimensional scaling plot of macroinvertebrate communities collected by Brown vacuum samples in July 2013 showing macroinvertebrate communities from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, Spring = spring-fed tributaries, Veg = marginal vegetation, Stress = 14.3, randomization p = 0.004 ...... 131

50. Canonical correspondence analysis biplot of macroinvertebrate community correlations to environmental variables in July 2013 from the Current and Jacks Fork rivers, Missouri. Arrow length indicates strength of correlation. Arrow direction indicates direction of relationship between variable and community. R = riffle, P = pool, V = marginal vegetation ...... 132

51. Two-way analysis of variance comparing temperature among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors ...... 133

52. Two-way analysis of variance comparing pH among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors ...... 134

53. Two-way analysis of variance comparing discharge among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences based on a square root transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors ...... 135

54. Two-way analysis of variance comparing depth among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences are based on a log

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transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors ...... 136

55. Two-way analysis of variance comparing embeddedness cover among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 137

56. Two-way analysis of variance comparing macrophyte cover among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences based on a square root transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 138

57. Two-way analysis of variance comparing organic cover among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences are based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 139

58. Two-way analysis of variance comparing substrate particle sizes among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 140

59. Two-way analysis of variance comparing velocity among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 141

60. Nonmetric multidimensional scaling plot of macroinvertebrate communities collected by Brown vacuum samples in July 2013 showing environmental variables between

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tributary types in the Current and Jacks Fork rivers, Missouri. Stress = 15.36, randomization p = 0.004 ...... 142

61. Nonmetric multidimensional scaling plot of macroinvertebrate communities collected by Brown vacuum samples in July 2013 showing environmental variables among mesohabitats in the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, Spring = spring-fed tributaries, Veg = marginal vegetation. Stress = 15.36, randomization p = 0.004 ...... 143

62. Three-way analysis of variance comparing taxonomic richness of the macroinvertebrate community comparing analyses when genera either excluded or included from analyses in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of Chironomidae, in Chiro = including genera of Chironomidae. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 168

63. Three-way analysis of variance comparing taxonomic richness of the macroinvertebrate community between tributaries in July and September 2013 from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of Chironomidae, in Chiro = including genera of Chironomidae. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 169

64. Two-way analysis of variance comparing taxonomic richness of Chironomidae between tributaries from the Current and Jacks Fork rivers, Missouri in July and September 2013. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Columns represent means of five replicates ± standard errors ...... 170

65. Three-way analysis of variance comparing Shannon’s Diversity Index values between tributaries in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of Chironomidae, in Chiro = including genera of Chironomidae. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 171

66. Three-way analysis of variance comparing biotic index values among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of

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Chironomidae, in Chiro = including genera of Chironomidae. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 172

67. Three-way analysis of variance comparing Stream Condition Index values between tributaries in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of Chironomidae, in Chiro = including genera of Chironomidae. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 173

68. Nonmetric multidimensional scaling of surface-fed and spring-fed tributary macroinvertebrates excluding genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 4.13, randomization p = 0.040 ...... 174

69. Nonmetric multidimensional scaling of surface-fed and spring-fed tributary macroinvertebrates including genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 9.21, randomization p = 0.016 ...... 175

70. Nonmetric multidimensional scaling of surface-fed and spring-fed mesohabitat macroinvertebrates excluding genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 16.46, randomization p = 0.004 ...... 176

71. Nonmetric multidimensional scaling of surface-fed and spring-fed mesohabitat macroinvertebrates including genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 16.61, randomization p = 0.004 ...... 177

72. Nonmetric multidimensional scaling of surface-fed and spring-fed mesohabitat genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 14.7, randomization p = 0.004 ...... 178

73. Two-way analysis of variance comparing Chironominae densities among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences reflect a log transformation. Columns represent means of five replicates ± standard errors ...... 179

74. Two-way analysis of variance comparing Tanypodinae densities between tributaries in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences reflect a square root transformation. Columns represent means of five replicates ± standard errors ...... 180

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75. Two-way analysis of variance comparing Tanypodinae densities among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences reflect a square root transformation. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 181

76. Two-way analysis of variance comparing Orthocladinae densities among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences reflect a square root transformation. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels ...... 182

77. Most abundant Chironomidae collected in July and September 2013 from tributaries in the Current and Jacks Fork rivers, Missouri. T = Thienemannimyia, C/O = Cricotopus/Orthocladius...... 183

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ABSTRACT

Disturbance is a dominant force in streams, and macroinvertebrates have adapted by using specialized strategies to reach refugia. Refugia include stable environments with abundant shelter and food resources. In the Missouri Ozarks, spring-fed tributaries are possible refugia. To determine if spring-fed tributaries serve as thermal refugia, submergent bidirectional movement was measured from spring-fed and surface-fed confluences in the Current River, Missouri during winter and summer.

Macroinvertebrates were sampled to capture standing and bidirectional movement.

Macroinvertebrate communities in summer significantly differed and formed three groups: surface-fed tributaries, Current River main channel, and spring-fed tributaries as supported by nonparametric analyses. Spring-fed tributary macroinvertebrate communities were distinct, which suggests these tributaries are unlikely thermal refugia for macroinvertebrates. Because greatest community differences existed between tributary types, mesohabitats were investigated. Mesohabitats differed in community composition and taxonomic richness within functional feeding groups with marginal vegetation having high taxonomic richness. Chironomidae among mesohabitats were analyzed because of their high diversity and density in streams. At the genus level, the inclusion of chironomids in analyses did not alter bioassessment metrics although chironomids alone were able to differentiate among mesohabitats reflecting differences found by the entire macroinvertebrate community. The most abundant chironomid taxa related best to nutrients and sediments. Further work with Chironomidae at the species level could improve environmental assessment and interpretation. The mesohabitat scale

xvii was able to differentiate among macroinvertebrate communities and should be further investigated in the Ozarks.

xviii

TABLE OF CONTENTS

ACKNOWLEDGEMENTS ...... ii

LIST OF TABLES ...... iii

LIST OF FIGURES ...... vi

ABSTRACT ...... xvii

CHAPTER

1. Disturbance in Stream Ecology (Literature Review) ...... 1 Literature Cited ...... 10

2. Role of Stream Refugia in Macroinvertebrate Communities (Literature Review) ...... 12 Literature Cited ...... 20

3. Tributaries as Refugia for Benthic Macroinvertebrates in Ozark Streams ...... 22 Abstract ...... 22 Introduction ...... 23 Methods ...... 26 Results...... 30 Discussion...... 34 Literature Cited ...... 39

4. Macroinvertebrate Diversity among Mesohabitats in Ozark tributaries ...... 88 Abstract ...... 88 Introduction ...... 89 Methods ...... 92 Study sites...... 92 Sampling methods ...... 93 Macroinvertebrate metric analyses ...... 93 Macroinvertebrate community analyses ...... 94 Environmental analyses ...... 95 Results...... 95 Macroinvertebrate community descriptions ...... 95 Macroinvertebrate metric analyses ...... 96 Macroinvertebrate community analyses ...... 97 Environmental analyses ...... 98 Discussion...... 99 Literature Cited ...... 104

5. Chironomidae in the Ozarks and the Importance of Taxonomic Resolution ...... 144 Abstract ...... 144

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Introduction ...... 145 Methods ...... 147 Results...... 150 Discussion...... 152 Literature Cited ...... 158

APPENDICES

A. Macroinvertebrates collected in slack-Surber samples in July 2011 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples ...... 184

B. Macroinvertebrates collected in slack-Surber samples during January 2012 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples ...... 188

C. Macroinvertebrates collected in upstream movement samplers during July 2011 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples ...... 193

D. Macroinvertebrates collected in upstream movement samplers during January 2012 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples ...... 195

E. Macroinvertebrates collected in drift samplers during July 2011 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples ...... 197

F. Macroinvertebrates collected in drift samplers during January 2012 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples ...... 202

G. Macroinvertebrates collected July 2013 from surface-fed tributary mesohabitats in the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation. Values are sums of original data from three subsamples ...... 206

H. Macroinvertebrates collected July 2013 from spring-fed tributary mesohabitats in the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation. Values are sums of original data from three subsamples ...... 212

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I. Chironomidae collected in July and September 2013 from surface-fed tributary mesohabitats in the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation. Values are sums of original data from three subsamples ...... 217

J. Chironomidae collected in July and September 2013 from spring-fed tributary mesohabitats in the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation. Values are sums of original data from three subsamples ...... 220

VITA ...... 223

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CHAPTER 1

Disturbance in Stream Ecology (Literature Review)

Disturbance is the dominant force shaping stream communities (Resh et al. 1988).

It plays a role in every stream ecosystem and dictates macroinvertebrate community composition. An extreme example is a one in 10,000 year flood event that occurred in the Missouri Ozark Black River. This disturbance was powerful enough to move banks, restructure the stream bed, and reduce the number of species by 8% and populations by as much as 100% in some areas four months afterward (McCord et al. 2009). In a similar disturbance of Oregon streams, debris flow areas still had drastically impacted macroinvertebrate communities 8 years later, which increased diversity and decreased dominant taxa (Danehy et al. 2012). Other areas in Oregon streams that experienced floods but not debris flows had lower diversity and higher dominance 8 years later

(Danehy et al. 2012). The impacts of disturbance in streams are potentially long lasting, which indicates disturbance regimes are critical in order to predict macroinvertebrate community composition.

The impact of disturbance can have negative or positive effects on macroinvertebrate communities. Physical substrate movement in streams acts as the predominant force altering densities, taxonomic composition, and food resources (Reice

1985). In support for this, macroinvertebrate density, taxonomic richness, and food resources decreased as disturbance frequency increased in substrate disturbance experiments in New Zealand (Death 1996). Although abundance or densities of 1 invertebrates decrease with disturbance, beneficial effects are probable depending upon the disturbance area and intensity. For example, species richness of the benthic macroinvertebrate community on experimental tiles placed in the current can increase when larger areas on the tiles are disturbed and when the disturbance intensity is increased (McCabe and Gotelli 2000). Disturbance is a crucial concept in stream ecology because of the unpredictability of its occurrence and effect on the biota. Where disturbance is extreme as in desert streams, floods or droughts have unpredictable effects on the biota. Certain invertebrates may have morphologies appearing to favor survival and resistance, but in Sycamore Creek, Arizona no taxa possessed attributes favoring either low or high flow conditions (Boulton et al. 1992). Similarly, meta-analyses of floods revealed drastically different modes of recovery over different time intervals (Lake

2000). Physical disturbance in streams is a driver in stream ecology due to both its severity on the habitat and the unpredictable response by the community.

A clear definition of disturbance in stream ecology must be established in order to research this concept. White and Pickett (1985) defined disturbance as a discrete event beyond natural stream variation that must alter the physical environment, food resources, or biota. Lake (2000) did not specify that the event must be discrete in time, but it must have a similar effect as previously defined. The biota must be killed or removed, food resources depleted, and habitat degraded. Under these criteria, the impact from the disturbance event can be measured and quantified. Biomass of macroinvertebrates, primary production, or substrate movement all are ways in which a disturbance can be detected (Resh et al. 1988, Death 2003, Death and Zimmerman 2005). Greater specificity might be added by classifying the disturbance as a pulse, press, or ramp

2 depending on the temporal legacy of the disturbance when it has ended (Lake 2000).

Other variables also are important when considering how to define a disturbance, including cause and how it affects the stream community. Flood and debris flows in the northwestern U.S. often occur together, but each event has unique impacts. Floods will scour resources, redistribute substrate, and remove biota, whereas debris flows during floods will deposit excess litter, fine sediment, nutrients, and drastically damage riparian vegetation (Danehy et al. 2012). Disturbance is a complex concept even when defined.

Recently, disturbance definitions have distinguished between the disturbance event and the response to disturbance (Death 2010). This added specificity cautions researchers to carefully choose variables required to answer certain questions. For instance, measuring substrate movement in streams is helpful in defining a disturbance event but will be of little use in determining the response by the biota (Death 2010). Disturbance is still a relatively young field in stream ecology despite its long history as a predominant force shaping the community.

Biotic responses to disturbance are affected largely by disturbance regimes. For example, the predatory macroinvertebrate communities in coastal streams of Canada were best explained by a disturbance variable rather than by fish predation or other habitat measures (Sircom and Walde 2009). Further, predator exclusion experiments that include disturbance treatments best predicted community composition. Variables such as macroinvertebrate density and richness were most closely related to scour and fill disturbance simulations than to predator exclusions (Effenberger et al. 2011). Certain forms of disturbance will also affect the way biota respond. Flood events tend to decrease interspecific interactions. Conversely, drought causes mortality due to increases

3 in biotic interactions (Lake 2000). Furthermore, community responses are more severe during drought and require longer periods of time to recover than those following a flood which maintains connectivity and refugia (Boulton et al. 1992).

Sample area impacts measures of biotic response when conducting research following disturbance. For example, whereas a flood due to a reservoir break was severe and recorded as a one in 10,000 year flood immediately downstream, it was only measured as a one in 10 year flood farther downstream (McCord et al. 2009). Scale of the sample area also can impact measures of biotic responses because of environmental heterogeneity. Heterogeneity in the physical habitat invariably increased species richness and decreased any variability due to the disturbance over time (Brown 2007). Scale also impacts the ability to detect disturbance events. If the area of the disturbance is confined, then sampling numerous streams lowers the ability to detect the biotic response. Large- scale disturbances should be addressed by large-scale studies with many replicates.

Large-scale disturbances may be drastic and affect the biota for months or many years

(Brown 2007). In such instances, small-scale studies of the community can lead to erroneous conclusions. However, these disturbance responses can be investigated using remote sensing techniques to try and capture large-scale trends in the environment.

Caution should be used in these cases because in-stream variables are likely to still relate better to macroinvertebrate responses than would land cover or land use variables (Paller et al. 2014). Variables such as substrate size and morphology are critical variables for determining success in macroinvertebrate colonization, habitation, and foraging

(Robinson et al. 2011) and should be measured whenever possible.

4

Anthropogenic disturbances have not been well studied but also can have long- lasting effects. Disturbances such as water flow diversions or even climate change are linked with press or ramp type responses in the biota. Heavy metal pollutants, reservoirs, and channelization result in similar responses where the biota are affected more severely with time, reaching a final plateau that does not improve as long as the disturbance exists

(Lake 2000). Ramp disturbances continually degrade stream communities unless improvement occurs. Altered flow regimes will alter substrate and sediment transport, thereby affecting channel form by decreasing habitat heterogeneity and possible refugia for macroinvertebrates. Reservoirs stabilize flow but if structural breaks occur the scale and intensity of the disturbance would be drastically increased, making recovery difficult, such as the reservoir failure in the Black River which resulted in a one in 10,000 year flood (McCord et al. 2009). To alleviate flooding issues near urban populations and to meet agricultural needs, stream channelization is practiced to confine streams in predictable pathways. These channels increase stream flow 35–1000x higher than baselines, which makes recovery or habitation by macroinvertebrates nearly impossible

(Hax and Golladay 1998). Because anthropogenic disturbances are typically severe and long-lasting, a distinct model predicting the biotic response to these disturbances would be useful. The subsidy-stress model was proposed by Death (2010) to account for the most important variables affected, including light and nutrient levels. Stream history also affects the severity of the biotic response to disturbance. Streams with a history of frequent disturbances are less likely to recover as quickly from disturbance as those streams which have a stable history (Death 1996).

5

Effects from disturbance events can be difficult to predict using models. The

Intermediate Disturbance Hypothesis (IDH) predicts species richness in a community to be dependent on both disturbance intensity and an underlying competitive hierarchy

(Connell 1978). Superior competitors outcompete and eliminate inferior species in the absence of disturbance or in low disturbance regimes. Intense disturbance will reduce superior competitors and favor inferior, fast colonizing species. It is only at intermediate disturbance levels when both superior and inferior competitors will be able to coexist

(Connell 1978). Heavy criticism of the IDH results from a lack of research supporting a strong competitive hierarchy in stream macroinvertebrate communities (Resh et al. 1988,

McCabe and Gotelli 2000, Death 2010). Further, the IDH assumes the stream ecosystem exists at a state of equilibrium in the absence of disturbance. Streams tend to be in a constant state of recovery where invertebrate communities have low densities which makes biotic interactions unlikely and maximum richness in the system equally improbable (Death 2010). Research at small scales supports that highest intensity disturbance events provide maximum diversity, which contradicts predictions by IDH

(McCabe and Gotelli 2000). Another predictor of disturbance effects is Huston’s dynamic equilibrium model (McCabe and Gotelli 2000), which holds that maximum diversity could peak at any disturbance level because it not only depends upon a competitive hierarchy but also on population growth within the community. When populations are in favorable environments and able to grow rapidly competitive exclusion takes place, but it will not occur in populations suppressed by surrounding conditions

(McCabe and Gotelli 2000). Regional trends across U.S. streams did not support predictions by either the IDH or Huston’s model during disturbance (Resh et al. 1988).

6

Biotic or abiotic dominance in macroinvertebrate community diversity is debatable but could be explained by Peckarsky’s harsh-benign hypothesis (Death 2010) where harsh disturbance results in abiotic factors governing communities and benign conditions resulting in biotic factors governing communities. The patch dynamics concept is a better fit for streams by incorporating the random aggregations of physical and biotic stream attributes (Townsend 1989). Here, the result of disturbance on diversity is dependent upon spatial and temporal variation in the stream (Townsend 1989). Spatial heterogeneity and seasonally dependent species increase species niches and enable coexistence by many species. Additionally, disturbance should be perceived as a resetting mechanism to begin succession anew in the stream community (Townsend

1989). This patch concept appropriately captures the unpredictable nature of streams and generally explains how high diversity can be due to high environmental heterogeneity.

However, specific predictions based on quantifiable variables are lacking. The relationship between refugia patch configuration and community recovery is necessary to better solidify the applicability of the patch dynamics concept (Lake 2000). Criticism of the patch dynamics concept also results from the assumption that heterogeneity always provides a refuge for some species during disturbance. Patches of specific substrates support high resistance by certain taxa during disturbance as was the case during flow increases by 1000x (Hax and Golladay 1998). However, this is not always supported in research where disturbance sometimes uniformly affects all “patches” of a stream (Brown

2007), resulting in only predation pressure elimination in certain patches when disturbance is not present. Patch dynamics still includes assumptions of competitive displacement, which is rarely documented in stream communities (Death 2010). Overall,

7 each model proposed in stream ecology emphasizes the resetting mechanism of disturbance, which forces succession to begin again (Reice 1985, Death 2002, Brown

2007).

Macroinvertebrate movement is necessary because disturbance acts as a resetting mechanism. Movement may occur by two means: passively or actively (Lake 2000).

Passive displacement of macroinvertebrates during disturbance will either kill the organism or fortuitously relocate it into a favorable refuge to remain until conditions elsewhere become habitable (Lake 2000). Patches of refuge for a species can include habitats, seasons, or generations (Lake 2000). Active displacement during disturbance occurs near disturbance events when environmental cues indicate unfavorable conditions are increasing. Increased flow levels before floods cue some macroinvertebrates to move deeper into the substrate. Decreasing flow during drought cues some macroinvertebrates to emerge or enter resistant diapause stages. Regardless of the method of macroinvertebrate movement, the movement itself is critical to recolonization and recovery of the community after disturbance. Small-scale substrate studies revealed active movement of macroinvertebrates to a certain substrate type in which angular substrates were preferred over spherical or oblate shaped stones during high flows

(Holomuzki and Biggs 2003). Additional evidence supports that the type of mechanism of movement preferred by macroinvertebrates depends upon the type of flow present.

Specifically, preference for drift or crawling by macroinvertebrates varied throughout the year (Moser and Minshall 1996). Generally, communities recover faster from disturbance if they possess organisms with higher vagility (Moser and Minshall 1996,

Lake 2000).

8

Successful community recovery following disturbance will depend entirely on setting the correct goals for management of the stream. Management goals all must have a benchmark using quantifiable criteria. Recovery from disturbance will also depend upon the stream community that managers consider to be desirable. Recovery to conditions for human use or recreation does not necessarily indicate that any life should exist in the stream. However, ecologists seek to meet goals forming an established community. This suggests a low benchmark or a return to a given biomass in the stream.

An intermediate benchmark might be to restore invertebrate function in the stream, thereby promoting maximum feeding groups. The most difficult benchmark, but also the most anthropogenically unbiased, is to restore historical species richness, which would also recover aesthetics, quality, and function. All recovery or monitoring efforts require active research into the disturbance events present and how these interact with the stream community to better grasp natural variation, successional trends, and critical factors during recovery.

9

Literature Cited

Boulton, A. J., C. G. Peterson, N. B. Grimm, and S. G. Fisher. 1992. Stability of an aquatic macroinvertebrate community in a multiyear hydrologic disturbance regime. Ecology 73: 2192–2207.

Brown, B. L. 2007. Habitat heterogeneity and disturbance influence patterns of community temporal variability in a small temperate stream. Hydrobiologia 586: 93–106.

Connell, J. H. 1978. Diversity in tropical rain forests and coral reefs. Science 199: 1302–1310.

Danehy, R. J., R. E. Bilby, R. B. Langshaw, D. M. Evans, T. R. Turner, W. C. Floyd, S. H. Schoenholtz, and S. D. Duke. 2012. Biological and water quality responses to hydrologic disturbances in third-order forested streams. Ecohydrology 5: 90–98.

Death, R. G. 1996. The effect of patch disturbance on stream invertebrate community structure: The influence of disturbance history. Oecologia 108: 567–576.

Death, R. G. 2002. Predicting invertebrate diversity from disturbance regimes in forest streams. Oikos 9: 18–30.

Death, R. G. 2003. Spatial patterns in lotic invertebrate community composition: is substrate disturbance actually important?. Canadian Journal of Fisheries and Aquatic Sciences 60: 603–611.

Death, R. G. 2010. Disturbance and riverine benthic communities: What has it contributed to general ecological theory? River Research and Applications 26: 15–25.

Death, R. G., and E. M. Zimmermann. 2005. Interaction between disturbance and primary productivity in determining stream invertebrate diversity. Oikos 111: 392–402.

Effenberger, M., S. Diehl, M. Gerth, and C. P. Matthaei. 2011. Patchy bed disturbance and fish predation independently influence the distribution of stream invertebrates and algae. Journal of Animal Ecology 80: 603–614.

Hax, C. L., and S. W. Golladay. 1998. Flow disturbance of macroinvertebrates inhabiting sediments and woody debris in a prairie stream. American Midland Naturalist 139: 210–223.

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Holomuzki, J. R., and B. J. Biggs. 2003. Sediment texture mediates high-flow effects on lotic macroinvertebrates. Journal of the North American Benthological Society 22: 542–553.

Lake, P. S. 2000. Disturbance, patchiness, and diversity in streams. Journal of the North American Benthological Society 19: 573–592.

McCabe, D. J., and N. J. Gotelli. 2000. Effects of disturbance frequency, intensity, and area on assemblages of stream macroinvertebrates. Oecologia 124: 270–279.

McCord, S. B., W. J. Elzinga, C. G. Scott, and J. C. Pozzo, Jr. 2009. Impacts of a catastrophic flood on a southeastern Missouri (U.S.A.) stream. Journal of Freshwater Ecology 24: 411–423.

Moser, D. C., and G. W. Minshall. 1996. Effects of localized disturbance on macroinvertebrate community structure in relation to mode of colonization and season. American Midland Naturalist 135: 92–101.

Paller, M. H., S. C. Slerrett, T. D. Tuberville, D. E. Fletcher, and A. M. Grosse. 2014. Effects of disturbance at two spatial scales on macroinvertebrate and fish metrics of stream health. Journal of Freshwater Ecology 29: 83–100.

Reice, S. R. 1985. Experimental disturbance and the maintenance of species diversity in a stream community. Oecologia 67: 90–97.

Resh, V. H., A. V. Brown, A. P. Covich, M. E. Gurtz, H. W. Li, G. W. Minshall, S. R. Reice, A. L. Sheldon, J. B. Wallace, and R. C. Wissmar. 1988. The role of disturbance in stream ecology. Journal of the North American Benthological Society 7: 433–455.

Robinson, C. T., S. Blaser, C. Jolidon, and L. N. S. Shama. 2011. Scales of patchiness in the responses of lotic macroinvertebrates to disturbance in a regulated river. Journal of the North American Benthological Society 30: 374–385.

Sircom, J., and S. J. Walde. 2009. Disturbance, fish, and variation in the predatory insect guild of coastal streams. Hydrobiologia 620: 181–190.

Townsend, C. T. 1989. The patch dynamics concept of stream community ecology. Journal of the North American Benthological Society 8: 36–50.

White, P. S., and S. T. A. Pickett (eds). 1985. Natural disturbance and patch dynamics: an introduction. The ecology of natural disturbance and patch dynamics. Academic Press, San Diego.

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CHAPTER 2

Role of Stream Refugia in Macroinvertebrate Communities (Literature Review)

Stream refugia are critical resources for the persistence of macroinvertebrates during disturbances or other adverse conditions. They may be defined according to temporal duration and structural dependability. Refugia are areas that serve as recolonist sources following biophysical disturbance events (Sedell et al. 1990). More specifically, these areas can be a spatial habitat or environmental variable that promotes either resistance or resilience (Sedell et al. 1990). These definitions must also consider macroinvertebrate biology and the landscape scale. Complex-life refugia include several habitats because these macroinvertebrates require each during a separate life stage

(Robson et al. 2013). Use of several habitats may be driven by dietary source changes across seasons, which physically move necessary resources. Other refugia are defined by use within a single life stage. For example, within-habitat refugia are patches within a habitat that remain physically stable during a disturbance (Robson et al. 2013).

Macroinvertebrate occupation of these refugia is mediated by chance events rather than active and intentional movement efforts. For example, discharge within channelized and unchannelized streams can be equal, but because the unchannelized streams have physical complexity, macroinvertebrates are typically pushed by chance into a refugium

(Negishi et al. 2002). Between-habitat refugia are similar habitats near disturbed habitats that can provide recolonists (Robson et al. 2013). Unlike within-habitat refugia, these

12 refugia are more predictable within streams due to their reliable structural stability.

Common examples are the hyporheic zone or lateral stream channels. One of the more effective refugia supporting fast recovery rates are ark-type refugia. These refugia have attributes that make them suitable to a broad range of taxa within the ecosystem (Robson et al. 2013). Perennial spring-fed tributaries or confluence areas could be either within- habitat refugia or ark-type refugia. Macroinvertebrate community recovery rates and recovery pathways will depend ultimately upon the type of refugium providing recolonists (Palmer et al. 1995).

Preservation of only a single refugium type does not provide long-term survival of macroinvertebrate communities. Rather, pathways that connect refugia with varying parameters across the stream network are necessary to maintain sustainable macroinvertebrate populations (Robson et al. 2013). This is a necessity because full recovery requires the protection of the majority of species present in the stream (Robson et al. 2013). Therefore, full recovery after disturbance suggests the ark-type refugia are ideal, and ark-type refugia should be a priority in conservation efforts when protection of all taxa is not feasible. Spring-fed confluences may be ark-type refugia due to intersection of contrasting environmental conditions.

Refugia exist at different spatial scales and can be linked to fluctuations in the flow regime (Palmer et al. 1995). From the smallest to largest stream scales, stream refugia are present at the channel subunit, channel unit, channel section, or watershed

(Sedell et al. 1990). More specifically, at the smallest scale of channel subunit, large boulders are a common refugium. Channel units can be riffles, pools, woody debris, backwaters, or marginal vegetation. At the larger channel section scale multiple habitats

13 exist, including the hyporheic zone, upstream channels, or side channels such as those found in braided rivers. At the largest watershed scale, refugia are entire streams unaffected by a disturbance. Protection at the larger refugia scales ensures protection of more metapopulations and therefore more of the entire macroinvertebrate community.

When large, intense disturbances occur, large-scale refugia should be incorporated in research and management. As an example, impounded rivers with unsteady flow and thermal cycles should have channel section or watershed-scale refugia that include several free-flowing tributaries. During smaller or more predictable disturbances (e.g., spring rains), refugia may include substrate structural complexity at the channel unit or section scale. When refugium scales do not reflect disturbance scales, recovery and maintenance of macroinvertebrate populations will be unsuccessful (Palmer et al. 1995).

Hierarchical effects of large scales on smaller scales affect refugia scales.

Naturally, large-scale refugia encompass small-scale refugia. For instance, if a downstream channel is a critical refugium during intense disturbance, then during less intense disturbances small-scale refugia are likely to exist in this same downstream channel (Kilbane and Holomuzki 2004). Further, large-scale run habitats were refugia during floods for Ironoquia punctatissima (Walker). At a smaller scale, cobble substrates within these runs were a microhabitat refugium during lighter rain events (Kilbane and

Holomuzki 2004). In another example, small-scale pools that remain inundated during drought are refugia in arid prairie streams. However, these pools are insufficient during supraseasonal droughts when only large-scale headwaters and springs can serve as refugia for recolonization (Burk and Kennedy 2013). Here it becomes clear that entire tributaries can function as a refugium to macroinvertebrates. Thermal refugia in both

14 summer and winter seasons of large channels can exist in smaller tributaries (Meyer et al.

2007) due to the presence of shade from the riparian zone or groundwater inflows.

Tributary scale refugia are understudied despite evidence of fast recovery rates following severe disturbance events because of tributaries. Macroinvertebrate communities in prairies recovered in days after flooding because of perennial tributary and hyporheic refugia (Dodds et al. 2004). Overall, management and conservation goals should always strive to protect large-scale refugia in order to keep macroinvertebrate communities resilient.

Refugia are a critical component to the recovery and rehabilitation process of every stream. Macroinvertebrate community resilience increases recovery rates to predisturbance levels in both taxonomic diversity and densities. Resilience qualities are important because it is rare that taxa can physiologically or physically resist severe and frequent disturbance events (Burk and Kennedy 2013). An additional factor in resilience and recovery rates is macroinvertebrate vagility (Gjerløv et al. 2003). Drift removes macroinvertebrates from ideal habitats by great distances, which causes delays in recolonization. If streams have structural physicochemical complexity, macroinvertebrates forcibly drifting in the water current are more likely to be caught in debris or slow flowing sections nearer to the original and ideal habitat (Gjerløv et al.

2003). In complex streams such as this, recovery rates are shortened. When no refugia are present, recolonization rates are a direct result of the disturbance frequency because only vagile macroinvertebrates capable of long distance dispersal are selected. More complex and numerous refugia support greater stream resilience and recovery rates after a disturbance.

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Limitations on the types and abundance of refugia come from the environmental conditions. Most stream refugia are reliant upon the current and flow patterns. For instance, in streams that dry seasonally the primary source of recolonization comes from surrounding streams in the watershed. Refugia from either upstream or downstream sources via drift and upstream movement is a large contributor of recolonists during the periods of connectivity during rain events (Gray and Fischer 1981). Whereas flight is one mode of macroinvertebrate dispersal at larger scales, submergent drift and upstream movement also are possible. These submergent movements are sometimes unnoticeable unless measured directly after floods, in which case the drift to upstream movement ratio can be between 25:1 to 2:1 (Gray and Fischer 1981). Macroinvertebrates of perennially flowing streams usually use the hyporheos during drought disturbances. However, macroinvertebrates of intermittent streams utilize aerial dispersal in response to drying.

This is possible due to quick responses to the precursors of drought including increasing stream temperatures and increased toxicities. The macroinvertebrate Gammarus pulex

(Linnaeus) moves into the hyporheos when stream temperatures rise (Wood et al. 2010).

Based on studies of refugia and recovery, the disturbance history of streams and the environmental conditions are important for refugia presence and abundance.

Macroinvertebrate conservation and management goals cannot be outlined or achieved without accounting for refugia. Surprisingly, refugia are underrepresented in restoration and conservation plans. Greater effort must incorporate theories concerning environmental hierarchies and patch dynamics in stream ecology with flow and disturbance regimes (Lake et al. 2007). Further, life histories for the entire aquatic fauna of streams must be understood to obtain maximum ecosystem quality.

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Macroinvertebrates with rapid life cycles, diapausing life stages, and aerial stages are typical of small streams with intermittent flow (Huryn et al. 2008). However, protection of intermittent streams is difficult and complex given variable state laws and interpretations. As a result of this legal system, anthropogenic pollution and sedimentation are common threats to intermittent streams. Larger perennial streams have longer-lived macroinvertebrates that lack diapausing life stages or have poorer dispersal abilities (Huryn et al. 2008). These macroinvertebrate communities tend to harbor greater densities of taxa with more specialized food requirements. Taxa sensitive to water quality should be monitored in groundwater-dominant spring-fed streams because contamination or water withdrawal is a growing threat. The utilization of refugia during all life stages of macroinvertebrates across a range of disturbance severities requires further research to preserve appropriate stream attributes at the proper spatial scales. At small scales, often the substrate is the most critical physical attribute whereas larger substrates offer suitable refugia. Although protection of entire watersheds is ideal, it is typically infeasible. Protecting stream lengths that encompass heterogeneity is an adequate compromise between the interests of private landowners and conservation managers using available monitoring methods (Dodds et al. 2004). The Ogallala-High

Plains Aquifer in Oklahoma has lowered greatly due to water abstraction for agriculture.

As a result, many prairie streams are now nonexistent, which has extirpated entire species and potential recolonists needed in upstream habitats (Dodds et al. 2004). Protecting the stream channel with a large riparian buffer may be insufficient in intermittent streams, whereas an entire cluster of streams and temporary ponds would be a better safeguard to diversity and abundance because research suggests aerial dispersal is dominant in these

17 systems. An understanding of refugia context in stream environments is key in the management of threatened stream habitats.

In the Missouri Ozarks, macroinvertebrate refugia exist at various spatial scales.

Spring-fed tributaries are one possible refugium in the Ozarks. These streams have characteristics that can provide stability during seasonal and stochastic disturbances.

Stable temperatures of spring-fed tributaries can greatly moderate temperatures in downstream channels (Mugel et al. 2009). Other factors of spring-fed tributaries supporting use as refugia include the maintenance of above-freezing temperatures, lower macroinvertebrate competition, and fewer top predators (Cantonati et al. 2012). Previous research indicates spring-fed streams function as refugia. For example, during severe floods, streambeds in surface-fed streams are scoured and slough algae, but algae nearer spring-fed tributaries remain because flow is moderated by springs (Cantonati et al.

2012). Even when human impacts are profound within a watershed, the spring-fed streams are commonly the least impacted and are able to host sensitive aquatic taxa that represent a large proportion of the regional fauna (Cantonati et al. 2012).

Previous research indicates the spring-fed streams in the Missouri Ozark region act as a large-scale refuge to macroinvertebrates during environmental stress and disturbance. Springs typically include non-insect macroinvertebrates at the source of groundwater inflow (Glazier 1991) and thermally offer a refuge to glacial and endemic macroinvertebrates (Erman and Erman 1995, von Fumetti et al. 2006, Barquin and

Scarsbrook 2008). Stable flows because of springs make these habitats ideal as refugia to not only spring-specific species but to all macroinvertebrates during disturbances

(Barquin and Scarsbrook 2008). The thermal stability in springs makes these habitats

18 ideal to fish with specific thermal requirements (Sedell et al. 1990). However, thermal stability is also critical to macroinvertebrates as a form of disturbance. During seasonal temperature extremes, macroinvertebartes will use springs as a refuge to escape either summer heat or winter freezes (Meyer et al. 2007). Active movement is a way to determine habitat suitability as a refugium to macroinvertebrates. Submergent upstream movement is commonly observed for amphipods. For example, Gammarus pulex

(Linnaeus) and Crangonyx pseudogracilis Bousfield move upstream into spring-fed tributaries during summer drought. Passive movement by other macroinvertebrates is likely to occur in springs during disturbance (Stubbington and Wood 2013). The Current and Jacks Fork rivers are pristine streams that are classified as references for the region

(Doisy and Rabeni 2001, Bowles et al. 2007). Thermal moderation in the Current and

Jacks Fork rivers occurs due to several spring-fed tributary inputs (Mugel et al. 2009).

The spring-fed tributaries are also characteristically stable in flow throughout the year

(Mugel et al. 2009). These characteristics and the connections to the more unpredictable surface-fed tributaries make spring-fed tributaries probable refugia during seasonal thermal extremes.

19

Literature Cited

Barquín, J., and M. Scarsbrook. 2008. Management and conservation strategies for coldwater springs. Aquatic Conservation: Marine and freshwater ecosystems 18: 580–591.

Bowles, D. E., J. A. Luraas, L. W. Morrison, H. R. Dodd, M. H. Williams, G. A. Rowell, M. D. Debacker, J. A. Hinsey, F. D. Usrey, and J. L. Haack. 2007. Protocol for Monitoring Aquatic Invertebrates at Ozark National Scenic Riverways, Missouri, and Buffalo National River, Arkansas. Natural Resource Report NPS/HTLN/NRR-2007/009. National Park Service, Fort Collins, Colorado.

Burk, R. A., and J. H. Kennedy. 2013. Invertebrate communities of groundwater- dependent refugia with varying hydrology and riparian cover during a supraseasonal drought. Journal of Freshwater Ecology 28: 251–270.

Cantonati, M., L. Fürderer, R. Gerecke, I. Jüttner, and E. J. Cox. 2012. Crenic habitats for freshwater biodiversity conservation: Toward an understanding of their ecology. Freshwater Science 31: 462–480.

Dodds, W. K., K. Gido, M. R. Whiles, K. M. Fritz, and W. J. Matthews. 2004. Life on the edge: The ecology of great plains prairie streams. BioScience 54: 205–216.

Doisy, K. E., and C. F. Rabeni. 2001. Flow conditions, benthic food resources, and invertebrate community composition in a low-gradient stream in Missouri. Journal of the North American Benthological Society 20: 17–32.

Erman, N. A., and D. C. Erman. 1995. Spring permanence, Trichoptera species richness, and the role of drought. Journal of the Kansas Entomological Society 68: 50–64.

Gjerløv, C., A. G. Hildrew, and J. I. Jones. 2003. Mobility of stream invertebrates in relation to disturbance and refugia: A test of habitat template theory. Journal of the North American Benthological Society 22: 207–223.

Glazier, D. S. 1991. The fauna of North American temperate cold springs: Patterns and hypotheses. Freshwater Biology 26: 527–542.

Gray, L. J., and S. G. Fisher. 1981. Postflood recolonization pathways of macroinvertebrates in a lowland Sonoran Desert stream. American Midland Naturalist 106: 249–257.

Huryn, A. D., J. B. Wallace, and N. H. Anderson. 2008. Habitat, life history, secondary production, and behavioral adaptations of aquatic insects, pp. 55–103, In R. W. Merritt, K. W. Cummins, and M. B. Berg (eds): An introduction to the aquatic insects of North America, 4th ed. Kendall/Hunt, Dubuque.

20

Kilbane, G. M., and J. R. Holomuzki. 2004. Spatial attributes, scale, and species traits determine caddisfly distributional responses to flooding. Journal of the North American Benthological Society 23: 480–493.

Lake, P. S., N. Bond, and P. Reich. 2007. Linking ecological theory with stream restoration. Freshwater Biology 52: 597–615.

Meyer, J. L., D. L. Strayer, J. B. Wallace, S. L. Eggert, G. S. Helfman, and N. E. Leona. 2007. The contribution of headwater streams to biodiversity in river networks. Journal of the American Water Resources Association 43: 86–103.

Mugel, D. N., J. M. Richards, and J. G. Schumacher. 2009. Geohydrologic investigations and landscape characteristics of areas contributing water to springs the Current River, and Jacks Fork, Ozark National Scenic Riverways, Missouri. Report 2009-5138. United States Geological Survey. Reston, VA.

Negishi, J. N., M. Inoue, and M. Nunokawa. 2002. Effects of channelization on stream habitat in relation to a spate and flow refugia for macroinvertebrates in northern Japan. Freshwater Biology 47: 515–1529.

Palmer, M. A., P. Arensburger, P. S. Botts, C. C. Hakenkamp, and J. W. Reid. 1995. Disturbance and the community structure of stream invertebrates: Patch-specific effects and the role of refugia. Freshwater Biology 34: 343–356.

Robson, B. J., E. T. Chester, B. D. Mitchell, and T. G. Matthews. 2013. Disturbance and the role of refuges in Mediterranean climate streams. Hydrobiologia 719: 77–91.

Sedell, J. R., G. H. Reeves, F. R. Hauer, J. A. Stanford, and C. P. Hawkins. 1990. Role of refugia in recovery from disturbances: Modern fragmented and disconnected river systems. Environmental Management 14: 711–724.

Stubbington, R., and P. J. Wood. 2013. Benthic and interstitial habitats of a lentic spring as invertebrate refuges during supra-seasonal drought. Fundamental and Applied Limnology 182: 61–73.

Von Fumetti, S., P. Nagel, N. Scheifhacken, and B. Baltes. 2006. Factors governing macrozoobenthic assemblages in perennial springs in north-western Switzerland. Hydrobiologia 568: 467–475.

Wood, P. J., A. J. Boulton, S. Little, and R. Stubbington. 2010. Is the hyporheic zone a refugium for aquatic macroinvertebrates during severe low flow conditions? Fundamental and Applied Limnology 176: 377–390.

21

CHAPTER 3

Tributaries as Refugia for Benthic Macroinvertebrates in Ozark Streams

Abstract

In-stream refugia are critical to macroinvertebrate resistance and resilience during and after disturbance events. Moderate disturbance due to temperature extremes at tributary scales have been largely neglected in the macroinvertebrate literature. The goal of this study was to determine if spring-fed tributaries act as thermal refugia during cold and hot seasonal periods. Specific objectives were to quantify the amount of submergent bidirectional macroinvertebrate movement, describe macroinvertebrate communities from confluences, and to determine important environmental predictors of communities.

Macroinvertebrates were sampled during the middle of summer and winter from three surface-fed and three spring-fed confluences in the Current River, Missouri. These macroinvertebrate communities were collected from riffle/run habitats using slack-

Surber, drift, and upstream-movement samplers. From 170 taxa and 51,197 individuals, macroinvertebrate samples represented three groups in nonmetric multidimensional scaling (NMS) space: surface-fed tributaries, spring-fed tributaries, and Current River.

Spring-fed tributary macroinvertebrate communities were significantly different from those of all other locations in the summer season. Drift quantities in both seasons was higher in spring-fed tributaries than in surface-fed tributaries while upstream movement was extremely low. Stream temperature and velocity were the best environmental predictors of macroinvertebrate communities as supported by NMS correlations. The

22 uniqueness of spring-fed tributary macroinvertebrate communities and low upstream movement quantities suggests spring-fed tributaries do not act as a refuge during thermal extremes. Drift pathways could be critical to recovery and colonization in the Current

River. The differences among locations were likely a result of tributary environments, seasonal variation, and the composition of species pools.

Introduction

Refugia are critical habitat resources for macroinvertebrate persistence in freshwaters. Stream refugia for macroinvertebrates are physical or temporal spaces where macroinvertebrates experience fewer negative effects from a disturbance than are present in surrounding areas (Burk and Kennedy 2013). Access into refugia occurs by passive or active movement (Lake 2000). For example, evidence supports passive and active movement into backwaters or woody debris and under boulders during scour by floods (Lake 2000). Active movement across habitat types to reach refugia is common during drought and involves molting into a terrestrial life stage or migrating into the hyporheos or pholeteros (Lake 2000). Organic pollution severely disturbs macroinvertebrate communities and involves a slow recovery (Meade 2004). For example, a paper mill in Louisiana discharged effluent into a small tributary, which decimated populations (Vazquez 2013). Large-scale downstream sections of the main- stem channel were a refuge and source of recolonization enabling a fast recovery which might have otherwise taken months or years (Vazquez 2013). Although they are less documented, thermal differences also constitute a disturbance in streams. Sources of more extreme thermal disturbance result from river regulation and water extraction. For

23 example, increased temperatures due to unseasonal water releases from top zones of a

Colorado River reservoir decreased caddisfly abundance and diversity. Here, recovery was possible only by adult flight from adjacent streams (Voelz et al. 1994). Refugia stabilize communities during moderate disturbance and allow for more rapid recovery after disturbance by providing habitat for recolonists.

In order for stream macroinvertebrates to resist or recover from disturbances they must be able to locate a refuge and often this may be accomplished by movement during the time of environmental stress. Macroinvertebrates can move actively by crawling, climbing, burrowing, swimming, or flying (Minckley 1964, Kovalak 1976, Brittain and

Eikeland 1988, Bilton et al. 2001, Fenoglio et al. 2002, Meutter et al. 2007) either in the direction of flow (Robinson et al. 2004) or against it (Minckley 1964, Humphries 2002).

The mode of macroinvertebrate movement is dependent on a combination of life stage

(Wallace 1990) and environmental attributes such as presence of refugia (Borchardt

1993). Responses to intense disturbances such as flood and drought in the stream environment have been well studied (Gray and Fisher 1981; Resh et al. 1988; Wallace

1990; Lake 2000, 2003). Typical submergent movement in response to these events is through drift or migration into the hyporheos (Fenoglio et al. 2002). Fewer studies have investigated the prevalence of submergent upstream movement or movement in the absence of moderate disturbance. Further, studies investigating submergent movement at scales of tributaries have not been investigated thoroughly.

Two key variables controlling macroinvertebrate movement into refugia include velocity and temperature (Lake 2000, Stubbington et al. 2011). Stream velocity and temperature are critical variables because they are associated with numerous other

24 environmental attributes. For instance, higher velocities are related to lower stream temperatures, higher dissolved oxygen, larger substrate particle sizes, and lower primary productivity. Variables not accounted for by velocity or temperature include pollution events and disturbance history. During high stream velocities, refugia exist in patches of lower flow or lower shear stress (Lancaster and Hildrew 1993). These areas of low flow exist at various spatial scales. At the microhabitat scale, the spaces downstream of boulders or large woody debris are low-flow refugia. At the reach scale, marginal vegetation, pools, and backwaters represent low-flow refugia (Sedell et al. 1990). At larger than reach scales, in-stream flow refugia include side channels, tributaries, or the hyporheos (Stubbington et al. 2011, Burk and Kennedy 2013). Macroinvertebrate movement into these refugia depends upon availability, ease of access, and disturbance severity.

Perennially flowing spring-fed streams have stable flow and temperatures

(Williams 1991), which provide an ideal tributary refugium to macroinvertebrates during disturbance (Stubbington and Wood 2013). These spring-fed streams are numerous and accessible throughout the Missouri Ozark region. The likelihood of spring-fed tributaries being a large-scale refugium should increase as disturbance severity increases. Spring- fed streams have near constant flow, which is one of the most important variables shaping macroinvertebrate community composition (Smith and Wood 2002). Spring-fed streams of prairies are crucial refugia because macroinvertebrates can persist in the summer season in these perennially flowing reaches when riffles and pools become disconnected and thermally stressful (Burk and Kennedy 2013). Often a high faunal overlap between perennial springs and downstream main channels occurs, which further

25 suggests that spring confluences are a more important refugium for macroinvertebrates than are other tributary types (Smith and Wood 2002). Further, persistence in perennially flowing surficial waters should be preferred rather than in the hyporheos or by diapause because this refugium is less costly physically and metabolically to macroinvertebrates

(Burk and Kennedy 2013). Perennially flowing streams with both eucreanal and epirhithral habitats also offer more variation in habitat form than do neighboring surface- fed streams (Cantonati et al. 2012). Flow and thermal stability combined with habitat complexity help support a dense and more diverse macroinvertebrate community in perennial springs than in surface-fed streams (Alvarez and Pardo 2007, Cantonati et al.

2012), which suggests spring-fed streams could be critical refugia.

The goal of this study was to determine if spring-fed tributaries act as large-scale submergent refugia to macroinvertebrates given stable temperatures and flow. Objectives were to quantify submergent bidirectional macroinvertebrate movement, describe macroinvertebrate communities at confluences, and determine critical environmental variables predicting the macroinvertebrate community composition.

Methods

The Current River is in the Salem Plateau area of the Ozark region and is characterized by thin soils and karstic Cambrian and Ordovician age sedimentary rock

(Mugel et al. 2009). The Ozark National Scenic Riverways, located in Carter, Dent, and

Shannon counties, has been administered by the National Park Service since 1964 and currently protects 134 miles of the Jacks Fork and Current river corridors, which amounts to 5% of the total watershed (Bowles et al. 2007, Mugel et al. 2009). Many surface-fed

26 streams (e.g., Big Creek, Rocky Creek, and Sinking Creek) are located on private lands with only the lowermost tributary reaches included within park boundaries. Similarly, most of the area in spring recharge zones is located beyond park boundaries with spring outflows located within the park (Mugel et al. 2009). Grazing, gravel mining, lead and zinc mining, timber harvest, and livestock operations occur within the oak-hickory dominated watershed (Bowles et al. 2007). Spring inflows account for 92% of the discharge of the Current and Jacks Fork rivers, which make up the Ozark National Scenic

Riverways. Among sampled springs, Blue Spring has a discharge of 7.75 m3 s-1, Pulltite

Spring 4.70 m3 s-1, and Round Spring 2.86 m3 s-1 (Mugel et al. 2009). Stream temperatures are largely influenced by spring inflows and more stabilized stream temperatures exist downstream of these springs (Mugel et al. 2009).

Macroinvertebrates were sampled and environmental variables measured in July

2011 and January 2012 at six tributary confluences within the Current River in Dent and

Shannon counties, Missouri (Fig. 1). Confluences are referred to as tributary types based upon whether the inflowing tributary is surface-fed or spring-fed. Of six tributaries sampled, three were spring-fed (Blue Spring, Pulltite Spring, and Round Spring) and three were surface-fed [Big Creek (West), Rocky Creek, and Sinking Creek]. Three locations were sampled within each tributary type in riffle-run habitats (Fig. 2): within the tributary (T), upstream of the tributary confluence in the Current River (UCR), downstream of the tributary confluence in the Current River (DCR). Within each location, three subsamples were collected. Grids were overlaid on each location and random numbers were used to determine where subsamples were collected.

27

Macroinvertebrates were collected using three sampling methods: slack-Surber, upstream-movement, and drift. The slack-Surber sampler (Bowles et al. 2007) quantitatively samples 0.25 m2 of the benthos following two-minutes of constant substrate disturbance by garden rake where invertebrates are collected in a 500 m mesh net (Fig. 3). Upstream-movement samplers (Fig. 4) and drift nets (modified from Hobbs and Butler 1981) (Fig. 5) sampled macroinvertebrates from the lower water column near the substrate. Both upstream-movement and drift sampler macroinvertebrate densities

(invertebrates 100 m-3) were calculated using the equation by Hauer and Lamberti (1996) where N = number of macroinvertebrates, t = time sampler was in stream, W = sampler width, H = sampler height, V = mean water velocity upstream of sampler.

Movement = (N)(100) (t)(W)(H)(V)(3600 s hr-1)

Nonparametric multivariate techniques were used to compare macroinvertebrate communities between tributary types and among locations. Taxa-by-site matrices were used in nonmetric multidimensional scaling (NMS) to find the lowest stress two- or three-dimensional plot representing greatest distance between tributary types or among locations (PC-ORD, version 6). In NMS space, greater distances between points indicate greater dissimilarities between communities. A Sorensen distance measure was used to configure plots in multidimensional space using either two or three axes depending on three autopilot trial runs. Plots were the result of random starting coordinates and 1000 runs. Plot significance was determined by randomizing data and comparing new configuration divergence to original configurations with 250 runs and 500 iterations to determine plot stress. Stress values less than 15 reflected plots representative of true

28 multidimensional distances among taxa (in reference to ecological data). Taxonomic significance was tested by the nonparametric multi-response permutation procedure

(MRPP) comparing locations as grouping variables across numerous permutations. In

MRPP, taxonomic comparisons among locations were tested using distance across multidimensional space and the standardized test statistic (T) with effect size (A)

(McCune and Grace 2002, Peck 2010).

Macroinvertebrate movement was analyzed using total density, individual taxon density, and direction. Movement was quantified and tested across tributary types and locations using a split-plot design (SAS 9.4). Tested individual taxonomic densities included the most abundant taxa (i.e. taxa composing ≥ 1% of sample). Comparisons of movement direction were made using three-way ANOVAs with tributary type, location, and direction as factors with locations embedded within tributary types. Normality was determined by Shapiro-Wilks test, homogeneity of variance by plots of residual variance, and significance was determined using  = 0.05. Pairwise comparisons were made using t-tests with Bonferroni adjustments.

Environmental variables were measured to characterize the stream where benthic subsamples were taken (Tables 1, 2). Percentage cover by macrophytes, periphyton, filamentous algae, organics, and embeddedness within the slack-Surber area were used to approximate substrate components visually. Upper benthic substrate particle sizes were estimated from five substrate grabs and use of the Wentworth size scale (Bowles et al.

2007). Water was characterized by depth, velocity, and discharge at the location.

Additional stream measures included temperature, dissolved oxygen, pH, and turbidity

(Tables 1, 2). All environmental variables, excluding percentage cover and discharge,

29 were measured directly upstream of the slack-Surber sampler. Discharge was approximated across the entire tributary or across the samplable width in the Current

River (including the entire stream at upper Current River sites). Environmental predictors of taxa were determined using nonparametric nonmetric multidimensional scaling (NMS) where the taxa matrix predicted environmental vectors using a Sorensen distance measure. Ordination starting coordinates were randomly selected. Solutions were based upon 1000 runs and 500 iterations. The significance of the solution was tested using randomized data with 250 runs (PC-ORD, version 6) (McCune and Grace

2002). Environmental parameter differences also were determined using two-way

ANOVA with the split-plot design where the tributary type and location were factors.

Normality was determined by Shapiro-Wilks test, homogeneity of variance by plots of residual variance, and significance was ascribed using  = 0.05, with t-tests for pairwise comparisons with Bonferroni adjustments.

Results

All totaled, 170 macroinvertebrate taxa were collected based on 51,197 individuals. Slack-Surber samples included 140 taxa based on 31,771 specimens representing 67 families and 19 orders (Appendices A, B). Upstream movement samples included 58 taxa based on 612 specimens representing 38 families and 12 orders

(Appendices C, D). Drift samples included 146 taxa based on 18,814 specimens representing 68 families and 18 orders (Appendices E, F).

The most abundant summer macroinvertebrates (those composing > 1% of sample densities) from all locations included Amnicola and Chironomidae (Fig. 6). Spring-fed

30 tributaries had significantly more Amnicola (p = 0.024) (Fig. 7) than did surface-fed tributaries; and Lepidostoma were significantly higher in surface-fed tributaries than in spring-fed tributaries (p = 0.018) (Fig. 8) (Table 3). Tricorythodes (p = 0.034) (Fig. 9) and Cheumatopsyche (p = 0.011) (Fig. 10) differed by locations with higher densities in the UCR and DCR locations than in the T location. Chironomidae differed among locations across tributaries with lowest densities in the surface-fed T location than remaining locations (p = 0.015) (Fig. 11). Ceratopsyche morosa (Hagen) was present in surface-fed tributary confluences but was not present in spring-fed tributary confluences.

Oligochaeta, Leucrocuta, Lepidostoma, and Stenacron were abundant in spring-fed tributary confluences but were not statistically higher than in surface-fed confluences

(Appendix A). Spring-fed T locations contained high densities of the amphipod

Gammarus, which could not be statistically tested due to ANOVA assumption violations

(Table 3).

The most abundant winter macroinvertebrates (those composing > 1% of samples) from all locations included Optioservus and Ephemerella (Fig. 12). Among the most abundant taxa, Psychomyia flavida Hagen were lowest in T locations (p = 0.010) (Fig.

13) (Table 4). Amnicola were highest in the spring-fed T location than in remaining locations (p = 0.007) (Fig. 14). Other taxonomic trends were evident but were not supported statistically. Gammarus pseudolimnaeus Bousfield was present in high densities in spring-fed tributaries. Although not statistically significant, Lepidostoma was more abundant in spring-fed confluences than in surface-fed confluences; oligochaetes and amphipods were more common in spring-fed tributaries than in surface-fed tributaries, and Plecoptera were more common in surface-fed tributaries (Appendix B).

31

Macroinvertebrate community similarity was related strongly to tributary type with variations due to the sampling season. Summer communities segregated in multidimensional space by locations. Spring-fed tributary communities included different compositions and proportions of taxa than did surface-fed tributaries (Fig. 15).

Three groups formed in NMS space when locations were compared: spring-fed T, surface-fed T, and UCR/DCR locations (Fig. 16). Within a single treatment, greatest taxonomic variation (distance between replicates in multidimensional space) existed among the surface-fed T locations (Sorensen distance = 0.90) and lowest taxonomic variation existed among spring-fed DCR locations (Sorensen distance = 0.53). Overall, summer macroinvertebrate communities were unique in the spring-fed T treatment with significant differences from all other sampled locations (T = -1.88, p = 0.042) (Table 5).

Winter macroinvertebrate communities from slack-Surber samples were less segregated by tributary type or treatment in multidimensional space. Tributary types were poorly differentiated from each other (Fig. 17). In general, only spring-fed T location replicates were more similar among one another than were any other location replicates (Fig. 18). Locations could not adequately predict winter macroinvertebrate communities (T = -1.10, p = 0.137) (Table 5).

Macroinvertebrate communities did not move in equal quantities in both stream directions. Summer and winter macroinvertebrate densities in drift densities were greater than densities in upstream movement samples (p < 0.001) (Table 6). Macroinvertebrates moving upstream in summer were extremely sparse and did not differ among locations

(Table 7). In winter, drifting macroinvertebrate densities were significantly higher in spring-fed tributaries than in surface-fed tributaries (p = 0.006) (Fig. 19) (Table 7).

32

Similar to summer trends, upstream movement in winter did not differ between tributary types (p = 0.108) or among locations (p =0.051) (Table 7).

Drifting summer macroinvertebrate communities were similar across tributary types (p = 0.379) (Fig. 20) and locations (p = 0.594) (Fig. 21) (Table 5). Drifting winter communities differed by tributary type (p = 0.001) (Fig. 22) and location (p = 0.023)

(Fig. 23). In pairwise comparisons of winter drift communities, spring-fed T locations were significantly different surface-fed T (p = 0.022), surface-fed UCR (p = 0.025), surface-fed DCR (p = 0.023), and spring-fed DCR (p = 0.042) locations. Surface-fed T location communities also differed from spring-fed DCR location communities (p =

0.025); all other pairwise contrasts were not significant (Table 5). Both surface-fed T and spring-fed T locations separated from the UCR and DCR locations in winter samples

(Fig. 23). NMS spatial groupings of winter drift were clearer between tributary types than by locations (Fig. 22). Due to matrix sparsity in the Sorensen distance matrix, upstream movement NMS plots could not be constructed reliably.

Environmental correlations to macroinvertebrate communities in multi- dimensional space varied with season. Stream temperature was most strongly correlated to benthic communities in summer (Fig. 24). Based on ANOVA analyses, temperature was reliant upon the interaction of tributary type with location (p < 0.001) (Table 8) (Fig.

25). Dissolved oxygen levels differed according to tributary type and location (p =

0.034) (Fig. 26), and the percent embeddedness cover differed by location (p = 0.035)

(Fig. 27) where T locations had highest embeddedness. Another important vector in

NMS space for distinguishing among summer locations was percent macrophyte cover

(Fig. 24).

33

Winter environmental variables poorly correlated with taxa (Fig. 28). Stream velocity was the best correlate followed by turbidity, pH, and stream temperature. Six significant differences in environmental variables between tributary types were detected in winter using ANOVA (Table 9). Between tributaries, spring-fed tributaries had higher temperatures (p = 0.002) (Fig. 29) and larger substrate sizes (p < 0.001) (Fig. 30).

Among locations, dissolved oxygen (p = 0.021) (Fig. 31) and pH (p = 0.032) (Fig. 32) were lowest in T locations. Interactions existed between tributaries and locations in percent macrophyte and percent periphyton cover. Macrophyte cover was highest in the spring-fed T location (p < 0.001) (Fig. 33) and periphyton cover was highest in the surface-fed DCR location, spring-fed UCR location, and spring-fed DCR location (p =

0.040) (Fig. 34).

Discussion

In-stream macroinvertebrate movement did not support the use of spring-fed tributaries in the Current River as refugia during moderate thermal disturbance. Evidence for recolonization at tributary scales is known only by drift rather than by upstream movement (Negishi et al. 2002). The strong current from spring-fed tributaries can partly explain why there was little upstream movement. Passive wash into the water column occurs regularly after which macroinvertebrates have great difficulty escaping the current to seek out refugia (Lancaster et al. 2006). Drift was important in samples, although drift distances were not measured in this study. Constant high discharge in most of the sampled locations and the high quantities of macroinvertebrates collected in drift samples suggested drift would be the only submergent option for tributary-scale movement into

34 refugia by macroinvertebrates. Thus, drifting macroinvertebrates appear to be the dominant means of submergent passive and active movement after disturbance in the

Current River.

Previous refugia work in streams indicates the refugium form depends heavily upon disturbance severity and stream size (Sedell et al. 1990). Thermal stress is understudied and as such it was difficult to know how stressful or severe this would be on macroinvertebrate communities. If seasonal thermal extremes were a low to moderate form of disturbance then the use of small-scale refugia nearby such as micro- or mesohabitats (Lancaster 2000) might be preferred. The Current River has a large and complex in-stream habitat, which increases the likelihood that a macroinvertebrate could enter a small scale refugium rather than needing to move greater distances to locate refuge tributaries. Deep pools represent a possible source of thermal refugia in the

Current River network. When temperatures are low in winter or high in summer, deep depths of pools are thermally moderated. A more severe form of thermal disturbance could have been necessary for tributaries to function as refugia. For example, dams disrupt natural physicochemical patterns to which macroinvertebrates have adapted.

However, below dams, macroinvertebrate persistence in the main channel occurs due to active movement from tributaries (Sedell et al. 1990). Further, stream temperatures at all sampled sites in the Current River of both seasons ranged between 2 and 27°C, which were within a range capable of supporting maximum macroinvertebrate richness and trophic levels (see Glazier 2012). The moderate to low thermal disturbance, strong flow throughout the year, and complexity at the micro- and mesohabitat scale of the Current

35

River suggests that macroinvertebrate movement into refugia is probably passive and reliant upon chance encounters with refugia.

Surface-fed T treatment replicates of macroinvertebrate communities were less precise taxonomically than were spring-fed T treatment replicates (evident in multivariate space). Distances among replicates of surface-fed tributaries in NMS plots were greater than were distances among spring-fed tributaries. Spring-fed streams have lower thermal variability which supports greater biotic specialization (Glazier 2012), including crustacean and gastropod taxa (Glazier 1991, Barquin and Death 2004) and lower taxonomic variation. In samples collected in the present study, crustaceans and gastropods made up between 41% and 68% of slack-Surber macroinvertebrate communities from spring-fed T locations. Thermal and flow stability in springs gives these non-emerging macroinvertebrates a competitive advantage over macroinvertebrates with an emergent stage (Glazier 1991, Clarke et al. 2008). Greater taxonomic precision among spring-fed T locations is also derived from lower taxonomic diversity in crustaceans and mollusks than in insects. In Missouri, only 78 species of crustaceans and gastropods have been documented, whereas 643 species of aquatic insects are known

(Sarver 2010). In addition, differences in precision of taxonomic communities could result from differences in recolonization rates. Spring-fed streams are known for slow recovery following disturbance (Barquin and Death 2004). The strong current tends to inhibit recolonization from submergent pathways and recolonization from aerial pathways is inhibited by the current washing eggs downstream (Glazier 1991).

Locations rather than tributary types were better predictors of macroinvertebrate communities. In some instances, taxonomic exchange resulting in increased similarity at

36 the tributary type would only occur after a severe flood event (Beckmann et al. 2005).

Water levels tend to dictate macroinvertebrate richness, similarity, and composition in a confluences (Mac Nally et al. 2011). However, samples in the Current River were targeted for low-flow periods in winter and summer rather than high flow or spate events.

If spring or autumn were targeted for sampling instead, it would be more likely that greater similarity at the tributary type level would have been discovered. Current River macroinvertebrate communities were taxonomically dissimilar across all sampled locations, indicating the absence of a high degree of taxonomic exchange. These results suggest also that movement might have been unnecessary given that there exists ample habitat for food resources, shelter, and physiological requirements. Until greater thermal stress can be measured and studied, it is unlikely that documentation of macroinvertebrate community composition will depend upon tributary types more than locations.

Macroinvertebrate community distances in multivariate space between tributary types was greater in summer than in winter. This difference is likely to result partially from phenological differences across seasons (see Harper et al. 2012). More taxa in winter could be in resting stages deep in the substrate reducing taxonomic differentiation.

Temperatures in winter also lowers macroinvertebrate metabolism and activity levels, which decreases the likelihood of capturing developed specimens that can be identified

(Grab 2014). Thus, the winter season usually has a lower detectable species pool, although this was not the case in the Current River. Winter macroinvertebrate communities from slack-Surber samples included 109 taxa and summer communities had

113 taxa. Summer environmental barriers among locations must have been great enough

37 to limit dispersal or habitats were more patchy and diverse to support patches of unique macroinvertebrate communities.

Tributary-scale refugia for macroinvertebrates during low thermal stress was not supported by movement samples. Instead, macroinvertebrate communities were likely a result of the influences from local species pools, environmental differences, and seasonal variations in locations. High quantities of drifting macroinvertebrates suggests recovery during low, moderate, and severe disturbance would be due to this mode of dispersal.

38

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Table 1. Environmental measures collected in July 2011 before slack-Surber samples were collected from tributaries of the Current River, Missouri. T = Tributary, UCR = upstream of tributary in Current River, DCR = downstream of tributary in Current River. Values are based on means from three subsamples.

Big Creek Rocky Creek Sinking Creek Blue Spring Pulltite Spring Round Spring Variable T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Dissolved Oxygen (mg L-1) 7.72 11.63 11.08 7.54 9.69 9.36 8.24 10.46 11.89 9.97 7.43 7.60 10.03 9.84 9.98 10.87 10.80 10.99 pH 7.1 7.6 7.6 7.6 7.6 7.8 7.3 7.1 7.3 7.4 7.5 7.7 6.9 7.2 7.3 6.8 7.4 7.4 Temperature (°C) 27.5 21.8 22.3 28.8 24.3 24.3 28.0 20.1 20.1 13.6 22.5 22.8 13.4 18.0 18.3 14.8 21.5 21.3

Embeddedness (%) 35.8 25.0 46.7 46.7 25.0 25.0 25.0 25.0 57.5 35.8 25.0 25.0 57.5 25.0 25.0 35.8 25.0 25.0 Filamentous Algae (%) 0.0 0.0 1.7 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 18.3 0.0 0.0 0.0 0.0 0.0 Macrophyte (%) 1.7 0.0 0.0 0.0 8.3 0.0 0.0 0.0 0.0 56.7 0.0 0.0 18.3 0.0 0.0 8.3 0.0 0.0 Organics (%) 25.0 25.0 25.0 10.0 16.7 5.0 11.7 18.3 18.3 5.0 5.0 5.0 18.3 22.5 5.0 5.0 25.0 18.3 Periphyton (%) 46.7 18.3 46.7 25.0 25.0 57.5 11.7 5.0 17.7 5.0 57.5 67.5 1.7 57.5 5.0 57.5 25.0 25.0 Substrate Size (Wentworth) 12.07 10.60 12.20 12.40 13.60 11.77 10.53 11.37 10.03 15.23 13.07 12.00 11.33 12.30 11.63 9.70 9.90 13.40

Depth (m) 0.13 0.19 0.23 0.27 0.60 0.34 0.26 0.30 0.38 0.22 0.24 0.26 0.35 0.21 0.27 0.54 0.34 0.62 43 3 -1

Discharge (m s ) 0.07 0.09 0.20 0.04 0.24 0.19 0.04 0.21 0.28 0.25 0.03 0.02 0.25 0.04 0.08 0.10 0.23 0.36 Samplable width (m) 2.5 6.5 7.5 2.2 2.0 0.0 7.0 3.0 5.0 11.0 8.5 4.0 10.0 5.0 5.0 7.0 9.0 11.0 Stream width (m) 3.0 18.5 22.5 2.2 56.0 48.0 22.0 43.5 39.5 11.0 71.0 58.5 10.0 42.0 47.0 13.5 48.0 50.5 -1 Velocity (m s ) 0.53 0.60 0.83 0.07 0.76 0.31 0.45 0.77 0.63 0.46 0.23 0.17 0.83 0.29 0.23 0.20 0.55 0.79

Table 2. Environmental measures collected in January 2012 before slack-Surber samples were collected from tributaries of the Current River, Missouri. T = Tributary, UCR = upstream of tributary in Current River, DCR = downstream of tributary in Current River. Values are based on means from three subsamples.

Big Creek Rocky Creek Sinking Creek Blue Spring Pulltite Spring Round Spring Variable T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Dissolved Oxygen (mg L-1) 11.9 14.3 13.4 13.1 13.6 13.1 13.5 11.9 12.3 10.6 12.8 12.9 10.4 13.3 11.7 10.2 11.9 12.8 pH 7.4 8.0 7.6 8.1 8.2 8.0 7.7 7.4 7.5 7.2 8.2 8.0 7.2 8.0 7.6 7.3 7.9 8.0 Temperature (°C) 4.7 5.4 5.1 6.7 9.0 8.6 2.9 5.7 2.8 11.1 7.2 7.0 11.8 8.6 5.5 11.4 9.5 8.8 Turbidity (NTU) 0.8 0.6 0.9 2.1 0.4 1.8 0.2 0.5 0.8 0.6 0.5 0.3 1.4 0.6 1.2 1.3 0.7 1.0

Embeddedness (%) 47.0 47.0 47.0 47.0 25.0 36.0 47.0 25.0 36.0 58.0 36.0 25.0 25.0 47.0 25.0 25.0 58.0 36.0 Filamentous Algae (%) 0.0 3.3 1.7 0.0 0.0 0.0 18.3 3.3 1.7 0.0 0.0 0.0 1.7 37.7 0.0 0.0 3.3 0.0 Macrophyte (%) 0.3 0.0 8.3 1.7 8.3 1.7 0.0 0.0 0.0 36.0 0.0 0.0 25.0 0.0 0.0 25.0 0.0 0.0 Organics (%) 16.7 25.0 25.0 25.0 10.0 25.0 18.3 25.0 18.3 25.0 25.0 25.0 18.3 25.0 25.0 29.3 25.0 25.0 Periphyton (%) 25.0 25.0 78.0 25.0 22.7 47.0 25.0 36.0 25.0 5.0 88.0 88.0 78.0 25.0 57.0 18.3 58.0 68.0 Substrate Size (Wentworth) 14.8 14.6 15.2 15.6 14.1 13.3 15 14.2 14.1 16.5 15.9 14.8 16.5 15.5 15.9 15.6 15.3 16.5

44 Depth (m) 0.17 0.22 0.20 0.30 0.36 0.39 0.21 0.21 0.35 0.33 0.18 0.27 0.21 0.15 0.37 0.43 0.31 0.51

Discharge (m3 s-1) 0.04 0.16 0.12 0.08 0.3 0.22 0.14 0.21 0.17 0.29 0.01 0.01 0.21 0.05 0 0.06 0.07 0.24 Samplable width (m) 5.5 8.0 7.8 2.2 2.3 3.7 9.3 6.7 6.0 2.5 7.7 4.2 6.8 4.2 3.7 5.0 16.5 3.0 -1 Velocity (m s ) 0.75 0.84 1.18 0.49 0.52 0.36 0.71 0.6 0.64 0.74 0.15 0.09 0.99 0.56 0.01 0.26 0.47 0.47

Table 3. Two-way analysis of variance tests of the most abundant taxa in July 2011 (> 1% of total sample) between tributaries or among locations in the Current River, Missouri. Dashes indicate data did not meet assumptions of ANOVA after transformation.

Taxa Effect NDF DDF F p Amnicola Tributary 1 12 6.67 0.024 * Location 2 12 3.77 0.054 Tributary x Location 2 12 2.48 0.126 Baetis Tributary 1 12 0.02 0.879 Location 2 12 1.00 0.396 Tributary x Location 2 12 1.52 0.258 Caenis Tributary 1 12 1.77 0.208 Location 2 12 1.69 0.226 Tributary x Location 2 12 3.36 0.070 Ceratopsyche slossonae Tributary - - - - Location - - - - Tributary x Location - - - - Cheumatopsyche Tributary 1 12 0.41 0.536 Location 2 12 6.80 0.011 * Tributary x Location 2 12 1.98 0.180 Chironomidae Tributary 1 12 8.11 0.015 Location 2 12 1.00 0.398 Tributary x Location 2 12 6.15 0.015 * Elimia Tributary 1 12 0.15 0.707 Location 2 12 1.10 0.364 Tributary x Location 2 12 1.17 0.344 Gammarus Tributary - - - - Location - - - - Tributary x Location - - - - Hydracarina Tributary - - - - Location - - - - Tributary x Location - - - - Isonychia Tributary 1 12 0.90 0.362 Location 2 12 0.34 0.718 Tributary x Location 2 12 0.63 0.550 Lepidostoma Tributary 1 12 7.57 0.018 * Location 2 12 0.82 0.506 Tributary x Location 2 12 0.71 0.512 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

45

Table 3. Continued.

Taxa Effect NDF DDF F p Maccaffertium Tributary 1 12 0.02 0.891 Location 2 12 3.27 0.074 Tributary x Location 2 12 0.81 0.466 Optioservus Tributary 1 12 2.02 0.181 Location 2 12 0.50 0.617 Tributary x Location 2 12 0.33 0.725 Plauditus Tributary 1 12 0.70 0.420 Location 2 12 0.37 0.700 Tributary x Location 2 12 0.27 0.769 Stenelmis Tributary 1 12 0.10 0.758 Location 2 12 1.49 0.264 Tributary x Location 2 12 0.59 0.568 Tricorythodes Tributary 1 12 0.00 0.961 Location 2 12 4.54 0.034 * Tributary x Location 2 12 0.90 0.433 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

46

Table 4. Two-way analyses of variance of the most abundant taxa in January 2012 (> 1% of total sample) between tributaries or among locations in the Current River, Missouri. Dashes indicate data did not meet assumptions of ANOVA after transformation.

Taxa Effect NDF DDF F p Amnicola Tributary 1 12 9.50 0.010 * Location 2 12 3.41 0.067 Tributary x Location 2 12 7.87 0.007 ** Baetis Tributary - - - - Location - - - - Tributary x Location - - - - Ceratopsyche piatrix Tributary 1 12 0.68 0.427 Location 2 12 1.01 0.393 Tributary x Location 2 12 0.57 0.583 Ceratopsyche slossonae Tributary 1 12 0.13 0.724 Location 2 12 1.64 0.235 Tributary x Location 2 12 0.44 0.654 Cheumatopsyche Tributary - - - - Location - - - - Tributary x Location - - - - Chironomidae Tributary 1 12 1.05 0.325 Location 2 12 2.28 0.145 Tributary x Location 2 12 1.23 0.328 Elimia Tributary 1 12 0.87 0.371 Location 2 12 0.34 0.719 Tributary x Location 2 12 0.46 0.641 Ephemerella Tributary 1 12 0.03 0.877 Location 2 12 1.54 0.254 Tributary x Location 2 12 0.03 0.968 Eurylophella Tributary 1 12 0.42 0.530 Location 2 12 3.78 0.053 Tributary x Location 2 12 1.75 0.215 Gammarus pseudolimnaeus Tributary - - - - Location - - - - Tributary x Location - - - - Hydracarina Tributary - - - - Location - - - - Tributary x Location - - - - * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

47

Table 4. Continued.

Taxa Effect NDF DDF F p Isonychia Tributary 1 12 0.16 0.700 Location 2 12 0.69 0.520 Tributary x Location 2 12 1.10 0.363 Lepidostoma Tributary - - - - Location - - - - Tributary x Location - - - - Maccaffertium Tributary 1 12 0.00 0.952 Location 2 12 2.36 0.137 Tributary x Location 2 12 3.09 0.083 Optioservus Tributary 1 12 0.01 0.931 Location 2 12 1.62 0.238 Tributary x Location 2 12 1.32 0.304 Psychomyia flavida Tributary 1 12 0.01 0.928 Location 2 12 7.04 0.010 * Tributary x Location 2 12 0.02 0.980 Pteronarcys Tributary - - - - Location - - - - Tributary x Location - - - - * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

48

Table 5. Multiple response permutation tests comparing macroinvertebrate community composition between tributaries and among locations in July 2011 and January 2012 from the Current River, Missouri.

Community Season Test T A p Drift Summer Tributary 0.002 0.000 0.379 Locations 0.323 -0.014 0.594 Winter Tributary -4.175 0.055 0.001 ** Locations -2.315 0.079 0.023 * Surface T v Spring T -2.908 0.183 0.022 * Surface T v Spring DCR -2.529 0.096 0.025 * Surface UCR v Spring T -2.446 0.091 0.025 * Surface DCR v Spring T -2.738 0.126 0.023 * Spring T v Spring DCR -1.638 0.036 0.042 * Surber Summer Tributary -0.916 0.011 0.168 Locations -1.878 0.058 0.042 * Surface T v Spring T -2.740 0.084 0.023 * Surface UCR v Spring T -2.724 0.121 0.023 * Surface DCR v Spring T -1.897 0.081 0.035 * Spring T v Spring UCR -2.300 0.122 0.028 * Spring T v Spring DCR -2.814 0.124 0.023 * Winter Tributary -0.195 0.002 0.346 Locations -1.098 0.035 0.137 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

49

Table 6. Three-way analysis of variance comparing macroinvertebrate movement direction (between drift and upstream movement samplers).

Season Effect DNF DDF F p Summer Tributary 1 24 3.36 0.079 Location 2 24 4.17 0.028 * Direction 1 24 341.87 <0.001 *** Tributary x Location x Direction 5 24 2.14 0.095 Winter Tributary 1 24 10.05 0.004 ** Location 2 24 5.40 0.012 * Direction 1 24 93.66 <0.001 *** Tributary x Location x Direction 5 24 0.51 0.763 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

50

Table 7. Two-way analysis of variance tests comparing macroinvertebrate quantities in movement samplers between tributaries and among locations in the Current River, Missouri. Mov. = movement.

Movement Effect NDF DDF F p Summer Drift Tributary 1 12 0.00 0.997 Location 2 12 2.00 0.178 Tributary x Location 2 12 2.22 0.152 Summer Upstream Mov. Tributary 1 12 3.18 0.100 Location 2 12 2.51 0.123 Tributary x Location 2 12 3.05 0.085 Winter Drift Tributary 1 12 10.85 0.006 ** Location 2 12 1.58 0.246 Tributary x Location 2 12 0.22 0.804 Winter Upstream Mov. Tributary 1 12 3.02 0.108 Location 2 12 3.84 0.051 Tributary x Location 2 12 0.61 0.559 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

51

Table 8. Two-way analysis of variance comparing environmental variables using tributary type and location as factors in July 2011 of the Current River, Missouri.

Variable Effect NDF DDF F p Depth Tributary 1 12 0.31 0.586 Location 2 12 0.20 0.818 Tributary x Location 2 12 1.11 0.360 Discharge Tributary 1 12 0.00 0.967 Location 2 12 0.68 0.525 Tributary x Location 2 12 2.49 0.124 Dissolved Oxygen Tributary 1 12 0.00 0.985 Location 2 12 1.36 0.293 Tributary x Location 2 12 4.55 0.034 * Embeddedness Tributary 1 12 0.73 0.410 Location 2 12 4.50 0.035 * Tributary x Location 2 12 3.03 0.086 Filamentous Algae Tributary 1 12 0.66 0.433 Location 2 12 0.83 0.460 Tributary x Location 2 12 1.17 0.343 Macrophytes Tributary 1 12 0.02 0.891 Location 2 12 0.51 0.615 Tributary x Location 2 12 1.64 0.234 Organics Tributary 1 12 1.64 0.225 Location 2 12 1.05 0.380 Tributary x Location 2 12 0.11 0.899 Periphyton Tributary 1 12 0.24 0.631 Location 2 12 0.41 0.675 Tributary x Location 2 12 1.35 0.297 pH Tributary 1 12 2.03 0.180 Location 2 12 2.85 0.097 Tributary x Location 2 12 0.40 0.677 Substrate Tributary 1 12 0.32 0.580 Location 2 12 0.00 0.997 Tributary x Location 2 12 0.17 0.846 Temperature Tributary 1 12 41.62 < 0.001 *** Location 2 12 0.11 0.895 Tributary x Location 2 12 23.55 < 0.001 *** Velocity Tributary 1 12 1.27 0.282 Location 2 12 0.28 0.758 Tributary x Location 2 12 1.50 0.262 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001 52

Table 9. Two-way analysis of variance comparing environmental variables between tributaries or among locations in January 2012 of the Current River, Missouri.

Variable Effect NDF DDF F p Depth Tributary 1 12 0.80 0.388 Location 2 12 2.09 0.166 Tributary x Location 2 12 1.03 0.386 Discharge Tributary 1 12 1.93 0.190 Location 2 12 0.02 0.979 Tributary x Location 2 12 4.25 0.040 * Dissolved Oxygen Tributary 1 12 10.64 0.007 ** Location 2 12 5.39 0.021 * Tributary x Location 2 12 3.02 0.087 Embeddedness Tributary 1 12 0.22 0.646 Location 2 12 0.72 0.506 Tributary x Location 2 12 2.72 0.106 Filamentous Algae Tributary 1 12 0.03 0.869 Location 2 12 1.16 0.348 Tributary x Location 2 12 0.92 0.424 Macrophytes Tributary 1 12 2.96 0.111 Location 2 12 12.22 0.001 ** Tributary x Location 2 12 18.57 <0.001 *** Organics Tributary 1 12 2.81 0.120 Location 2 12 0.22 0.802 Tributary x Location 2 12 0.13 0.877 Periphyton Tributary 1 12 7.08 0.021 * Location 2 12 12.39 0.001 ** Tributary x Location 2 12 4.27 0.040 * pH Tributary 1 12 0.20 0.665 Location 2 12 4.67 0.032 * Tributary x Location 2 12 3.37 0.069 Substrate Tributary 1 12 19.58 <0.001 *** Location 2 12 2.68 0.110 Tributary x Location 2 12 0.16 0.857 Temperature Tributary 1 12 14.60 0.002 ** Location 2 12 1.49 0.265 Tributary x Location 2 12 3.69 0.056 Turbidity Tributary 1 12 0.06 0.812 Location 2 12 1.41 0.281 Tributary x Location 2 12 0.28 0.759 Velocity Tributary 1 12 4.03 0.068 Location 2 12 0.78 0.482 Tributary x Location 2 12 1.44 0.275 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001 53

Sinking Creek

Big Creek Blue Spring Pulltite Spring Round Spring

Rocky Creek

Figure 1. Macroinvertebrate sampling sites along Current River, Missouri. Current River is shown in bold. Big Creek, Sinking Creek, and Rocky Creek are surface-fed tributaries flowing into the Current River. Pulltite Spring, Round Spring, and Blue Spring are spring-fed tributaries flowing into the Current River.

54

UCR

T Confluence

DCR

Figure 2. Current River locations within a stream confluence. T = tributary, UCR = upstream of tributary in Current River, DCR = downstream of tributary in Current River. Arrows indicate direction of flow. Circles represent the number of subsamples collected within locations from riffle/run habitat.

55

slack-Surber Sampler

Figure 3. Slack-Surber sampler to collect standing macroinvertebrate communities.

56

Figure 4. Upstream-movement sampler to collect macroinvertebrates crawling upstream.

57

Figure 5. Drift sampler to collect macroinvertebrates moving downstream.

58

20

18

16

14 (%)

12

10

8

6 Most abundant Most

4 macroinvertebrates 2

0

Figure 6. Most abundant macroinvertebrates collected from the Current River, Missouri from slack-Surber samples collected in July 2011.

59

350

) a 2 - 300

250

200

150

100 b

50 Density (individuals 0.05 m 0.05 (individuals Density

0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 7. Two-way analysis of variance comparing Amnicola density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

60

70 a

)

2 - 60

50

40

30

20 b

10 Density (individuals 0.05 m 0.05 (individuals Density 0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 8. Two-way analysis of variance comparing Lepidostoma density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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160 a

)

2 - 140

120

100 a

80

60

40 b

20 Density (individuals 0.05 m 0.05 (individuals Density 0 Surface T Spring T Surface UCR Spring UCR Surface DCR Spring DCR Locations

Figure 9. Two-way analysis of variance comparing Tricorythodes density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors. Note the different arrangement of axis labels.

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a

180

) 2 - 160

140

120

100

80

60 a

40

20 b Density (individuals 0.05 m 0.05 (individuals Density 0 Surface T Spring T Surface UCR Spring UCR Surface DCR Spring DCR Locations

Figure 10. Two-way analysis of variance comparing Cheumatopsyche density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors. Note the different arrangement of axis labels.

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400 a

) 350

2 -

300

250

200

150 a a

100 a a

Density (individuals 0.05 m 0.05 (individuals Density 50 b 0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 11. Two-way analysis of variance comparing Chironomidae density differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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20 18

(%) 16 14 12 10 8

Most abundant Most 6

4 macroinvertebrates 2 0

Figure 12. Most common macroinvertebrates collected from the Current River, Missouri from slack-Surber samples in January 2012.

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140

a

) 2

- 120 a

100

80

60

40

20 b Density (individuals 0.05 m 0.05 (individuals Density

0 Surface T Spring T Surface UCR Spring UCR Surface DCR Spring DCR Locations

Figure 13. Two-way analysis of variance comparing Psychomyia flavida density differences among locations in January 2012 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors. Note the different arrangement of axis labels.

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450 a

) 2

- 400

350

300

250

200

150

100 50 b Density (individuals 0.05 m 0.05 (individuals Density b b b 0 b Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 14. Two-way analysis of variance comparing Amnicola density differences among locations in January 2012 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Differences are based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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Tributary Types Surface-fed Spring-fed

+ Centroid NMS 2 NMS

NMS 1

Figure 15. Nonmetric multidimensional scaling plot of macroinvertebrate communities in July 2011 slack-Surber samples from the Current River, Missouri. Stress = 9.568, randomization p = 0.008.

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Locations

T Surface NMS 2 NMS UCR DCR

T Spring UCR DCR + Centroid NMS 1

Figure 16. Nonmetric multidimensional scaling plot of macroinvertebrate communities in July 2011 slack-Surber samples from the Current River, Missouri. Surface = surface- fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 9.568, randomization p =0.008.

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NMS 2 NMS

Tributary Types Surface-fed Spring-fed + Centroid NMS 1

Figure 17. Nonmetric multidimensional scaling plot of macroinvertebrate communities in January 2011 slack-Surber samples from the Current River, Missouri. Stress = 7.688, randomization p = 0.004.

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Locations

T Surface NMS 2 NMS UCR DCR

T Spring UCR DCR + Centroid NMS 1

Figure 18. Nonmetric multidimensional scaling plot of macroinvertebrate communities in January 2012 slack-Surber samples from the Current River, Missouri. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 7.688, randomization p = 0.004.

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) 100

1 a -

s 90

3 3 - 80

70

60

50

40 30 b 20

10

Quantity (individuals 100 m 100 (individuals Quantity 0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 19. Two-way analysis of variance comparing macroinvertebrate drift among locations in January 2012 from the Current River, Missouri. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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NMS 2 NMS

Tributary Types Surface-fed Spring-fed + Centroid NMS 1

Figure 20. Nonmetric multidimensional scaling plot of macroinvertebrate communities in July 2011 drift samples from the Current River, Missouri. Stress = 11.698, randomization p = 0.004.

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Locations

T Surface NMS 2 NMS UCR DCR

T Spring UCR DCR + Centroid NMS 1

Figure 21. Nonmetric multidimensional scaling plot for macroinvertebrate communities in July 2011 drift samples from the Current River, Missouri. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 11.698, randomization p = 0.004.

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NMS 2 NMS

Tributary Types Surface-fed Spring-fed + Centroid NMS 1

Figure 22. Nonmetric multidimensional scaling plot of macroinvertebrate communities in January 2012 drift samples from the Current River, Missouri. Stress = 10.906, randomization p = 0.004.

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Locations

T Surface NMS 2 NMS UCR DCR

T Spring UCR DCR + Centroid NMS 1

Figure 23. Nonmetric multidimensional scaling plot of macroinvertebrate communities in January 2012 drift samples from the Current River, Missouri. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 10.906, randomization p = 0.004.

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NMS 2 NMS

NMS 1

Figure 24. Nonmetric multidimensional scaling biplot of macroinvertebrate community correlations with environmental variables in July 2011 slack-Surber samples from the Current River, Missouri. Arrow length indicates strength of correlation. Arrow direction indicates increase environmental values. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 9.568, randomization p = 0.004.

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30 a

25 b b

b b C)

20 c 15

10 Temperature ( Temperature 5

0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 25. Two-way analysis of variance comparing temperature differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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14

12 a a )

1 a - ab ab 10 b 8

6

4

Dissolved Oxygen (mg L (mg Oxygen Dissolved 2

0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 26. Two-way analysis of variance comparing dissolved oxygen differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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60 ab a

50 (%) 40 b 30

20 Embeddedness 10

0 Surface T Spring T Surface UCR Spring UCR Surface DCR Spring DCR Locations

Figure 27. Two-way analysis of variance comparing embeddedness cover differences among locations in July 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors. Note the different arrangement of axis labels.

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NMS 2 NMS

NMS 1

Figure 28. Nonmetric multidimensional scaling biplot of macroinvertebrate community correlations with environmental variables in January 2012 slack-Surber samples from the Current River, Missouri. Arrow length indicates strength of correlation. Arrow direction indicates increase environmental values. Surface = surface-fed tributary type, Spring = spring-fed tributary type, T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of the tributary confluence in the Current River, stress = 7.688, randomization p = 0.004.

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14 a 12

10 C)

b 8

6

4 Temperature ( Temperature

2

0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 29. Two-way analysis of variance comparing temperature differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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18 b a

16

14

12

10

8

6

4

Substrate size (Wentworth) size Substrate 2

0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 30. Two-way analysis of variance comparing substrate size differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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16 a

b a )

1 14 -

12

10

8

6

4

2 Dissolved oxygen (mg L (mg oxygen Dissolved 0 Surface T Spring T Surface UCR Spring UCR Surface DCR Spring DCR Locations

Figure 31. Two-way analysis of variance comparing dissolved oxygen differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors. Note differences in arrangement of axis labels.

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a 8.2 b ab

8

7.8

7.6

7.4 pH

7.2

7

6.8

6.6 Surface T Spring T Surface UCR Spring UCR Surface DCR Spring DCR Locations

Figure 32. Two-way analysis of variance comparing pH differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors. Note differences in arrangement of axis labels.

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35 a

30

25 (%)

20

15

10 Macrophytes b b 5 b b b

0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 33. Two-way analysis of variance comparing macrophyte cover differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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90 a a 80

70 a

(%) 60

50

40 b 30 b Periphyton b 20

10

0 Surface T Surface UCR Surface DCR Spring T Spring UCR Spring DCR Locations

Figure 34. Two-way analysis of variance comparing periphyton cover differences among locations in January 2011 from the Current River, Missouri. T = tributary flowing into Current River, UCR = upstream of tributary confluence in the Current River, DCR = downstream of tributary confluence in the Current River. Pairwise comparisons with Bonferroni adjustments. Columns represent means of three replicates ± standard errors.

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CHAPTER 4

Macroinvertebrate Diversity among Mesohabitats in Ozark Tributaries

Abstract

Structural heterogeneity and biological diversity within a stream is captured best by sampling multiple mesohabitats. Functional feeding groups (FFGs) reflect food resource availability and can be used to interpret resources at the mesohabitat level. The goal of this study was to document macroinvertebrate communities from physically comparable mesohabitats in surface-fed and spring-fed tributaries. To address this goal, the environmental characteristics of mesohabitats and functional use were analyzed. In the summer of 2013, macroinvertebrates from 10 tributaries in the Current and Jacks Fork rivers, Missouri were sampled using a Brown vacuum sampler from riffle, pool, and marginal vegetation mesohabitats. From 144 taxa and 20,826 individuals, macroinvertebrate communities from the three mesohabitats and two tributary types were characterized. Tributary types and mesohabitats significantly differed in macroinvertebrate communities as supported by nonmetric multidimensional scaling.

Among FFGs, collectors were taxonomically richer in surface-fed mesohabitats than in spring-fed mesohabitats. Filterers were richer in riffles than in pool mesohabitats.

Scraper taxonomic richness was higher in riffles than in either pool or marginal vegetation mesohabitats. Taxonomic dominance by Chironomidae was higher in spring- fed tributaries than surface-fed tributaries; and diversity was higher in riffle and marginal mesohabitats than in pools. Mesohabitats could be differentiated by multiple 88

environmental variables including temperature, discharge, pH, depth, embeddedness, organics, macrophytes, velocity, and substrate size using canonical correspondence analysis. Macroinvertebrate communities at the mesohabitat level could be differentiated by multivariate techniques and taxonomic richness within FFGs. Further, tributary types were distinct and were likely a reflection of differences in disturbance regimes according to flow. Multimesohabitat studies are critical in ecology for understanding community relationships with habitat structure at small scales.

Introduction

Stream tributaries provide downstream macroinvertebrate communities with a more diverse array of species due to the greater habitat heterogeneity present after they converge. The inflowing tributaries have a higher connectivity to the watershed than do downstream main-stem channels (Freeman et al. 2007). As such, headwaters and small tributaries act as the mouths to the watershed by incorporating terrestrial nutrients and inorganic materials, which then support the downstream aquatic community (Clarke et al.

2008). For instance, perennial headwaters are responsible for 65% of the nitrogen flux in lotic systems (Alexander et al. 2007) and the bulk of organic matter transport and nutrient cycling (Clarke et al. 2008). Further contributions include structural variation.

Tributaries and small tributary confluences have higher diversity because physical complexity is higher than that of the downstream network (Finn et al. 2011). Hydraulic and substrate size heterogeneity also are high and are critical attributes in macroinvertebrate environments that favor increased colonization (Rice et al. 2001).

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Overall, macroinvertebrate taxonomic diversity is high in stream networks because of these small tributaries and is reflected by high -diversity values (Clarke et al. 2008).

Within a tributary are multiple mesohabitats that are distinct in physicochemical attributes and in corresponding macroinvertebrate communities. The array of mesohabitats increases in tributaries when compared to main stems (Rice et al. 2006) in part because of a greater land-stream interface (Freeman et al. 2007). Added complexity is a result of disturbance, which can be more frequent and severe in tributaries because of a higher surface area to volume ratio (Resh et al. 1988). Niche availability increases in tributaries because leaf litter and woody debris inputs create mesohabitats through snags of sediment, which also decrease velocities (Gomi et al. 2002). Potential refugia are present when there are more mesohabitats in tributaries, which creates greater resilience potential against disturbance. For example, sediments and slack-water areas in the margins of streams are favorable for macrophyte growth. These macrophyte beds are a complex substrate for macroinvertebrates and offer shade, shelter from predators, and plentiful food resources (Epele et al. 2012).

Macroinvertebrate bioassessment and ecological interpretations depend upon sampling protocol. Macroinvertebrate samplers are designed for specific habitats. Most assessments are based upon a single mesohabitat, usually riffles, and one sampler type, the Surber sampler, to maximize efficiency and allow for greatest comparability with other regional studies (Clarke et al. 2008). However, from an ecological perspective, it is valuable to perform more extensive studies to fully characterize the stream fauna, document new species, and provide a foundation for understanding biological interactions. The most stable and often the most biologically diverse mesohabitat is

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marginal vegetation (Armitage and Cannan 2000, Epele et al. 2012), whereas the most disturbed but least densely occupied mesohabitat are riffles (Costa and Melo 2008).

Therefore, studies of marginal vegetation are critical to understanding disturbance and community recovery. Despite the benefit of including multiple habitats, sampling a diverse array of mesohabitats is discouraged because of the need for comparability among studies. The interpretation of stream community changes over extended time periods and can be compromised by inconsistent inclusion of mesohabitats. Although bioassessments are streamlined by minimizing sample diversity, caution must be used when interpreting the implications of habitat value based upon records that were biased towards single mesohabitats.

Expanding sampling protocols will increase the likelihood of describing a more comprehensive macroinvertebrate community that reflects the true members in the environment (Clarke et al. 2008). In turn, this increases the ability to differentiate among sites in meta-analyses (Cao et al. 2002). Not surprisingly, small streams have been found to be extremely diverse due in part to long-term research studies that combined exhaustive protocols to understand the entire stream. For example, in the Breitenbach stream, a first order stream in Germany studied for 25 years, 1,004 invertebrate taxa were found (Allan and Castillo 1995). Although the most common stream mesohabitat studied is the riffle, a more complete understanding of the entire benthos, individual interactions, and community stability relies upon a more complex and intensive investigation of all mesohabitats.

The goal of this study was to describe how physically comparable mesohabitats from surface-fed and spring-fed Ozark tributaries were differentially occupied by benthic

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macroinvertebrates. Specific objectives included an examination of the complete community in each tributary type, a physical comparison among mesohabitats within and across tributary types, and a functional classification of macroinvertebrate communities within mesohabitats.

Methods

Study sites

Macroinvertebrates were sampled in July (one site in September due to rain) 2013 from 10 tributaries flowing into the Current or Jacks Fork rivers in Carter, Dent, and

Shannon counties, Missouri. Tributaries were selected based on dominant groundwater inflow being classified as either spring-fed (groundwater dominant) or surface-fed (non- groundwater dominant). Tributary site choice also was limited by the availability of three comparable mesohabitats. Sampled sites included both surface-fed [Ashley Creek, Bay

Creek, Big Creek (West), Rocky Creek, and Sinking Creek] and spring-fed (Alley Spring,

Blue Spring, Cave Spring, Pulltite Spring, and Round Spring) tributaries (Fig. 35).

Mesohabitats sampled were marginal vegetation, pools, and riffles. These mesohabitats were selected to maximize differences in stream velocity and depth. More specifically, riffles had the fastest stream velocities, shallowest depths, fewest macrophytes, and largest substrate sizes. Pools had slow to zero stream velocity, deepest water, few macrophytes, abundant detritus, and small substrate sizes. Marginal vegetation had slow to zero stream velocity, shallow water, highest macrophyte cover, and small substrate sizes (Cluer and Thorne 2013).

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Sampling methods

Macroinvertebrates were randomly sampled from all three mesohabitats with a

Brown vacuum sampler (Fig. 36) (Brown et al. 1987). The sampler was run for five minutes while disturbing substrate with a rake for each sample. Macroinvertebrates were collected using a 500 m nylon mesh from an area of 0.05 m2. All specimens were preserved in 80% ethanol and later processed in the laboratory. Macroinvertebrates were identified to the lowest practical level using national and regional keys (e.g., Poulton and

Stewart 1991, Moulton and Stewart 1996, Smith 2001, Merritt et al. 2008). Samples were processed by subsampling using National Park Service protocols (Bowles et al.

2007). After identification, macroinvertebrates were classified into five groups according to functional feeding groups: collectors, filterers, predators, scrapers, and shredders

(Cummins et al. 2008).

Macroinvertebrate metric analyses

Macroinvertebrate communities were characterized using both taxonomic composition and functional feeding groups (FFGs). Taxonomic composition of tributaries and mesohabitats were compared using taxonomic richness (TR), Shannon’s

Diversity Index (H’), densities, dominance (D), taxonomic richness within FFGs, and densities within FFGs in split-plot design two-way analysis of variance (ANOVA) with tributary type and mesohabitat as factors (SAS, version 9.4). Heterogeneity of variance was checked using residual variance plots and normality using Shapiro-Wilks test. If assumptions of ANOVA were not met, data were square root or log transformed. Where statistical significance existed  = 0.05), pairwise comparisons were performed using t-

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tests with Bonferroni adjustments. General relationships among tributaries and mesohabitats were supported with Jaccard’s similarity score (J) comparisons.

Macroinvertebrate community analyses

Macroinvertebrate community differences in taxonomic composition also were assessed in multidimensional space using nonmetric multidimensional scaling (NMS) where tributaries and mesohabitats were expected to form unique clusters (PC-ORD, version 6). In NMS, greater distances between points indicates greater dissimilarities between communities. Taxa-by-site matrices were assessed with NMS to find the lowest stress in two- or three-dimensional plots that maximize distance between tributaries and among mesohabitats. Sorensen similarities among groups were calculated to form a distance measure to configure plots in multidimensional space using either two or three axes depending on three autopilot trials (Peck 2010). Plots were the result of random starting coordinates and 1000 runs. Plot significance was determined by randomizing data and comparing new configuration divergence to original configurations with 250 runs and 500 iterations to find stress. Stresses lower than 15 reflect plots representative of true multidimensional distances in ecological data (McCune and Grace 2002).

Significance of taxonomic composition distinction was tested by a multi-response permutation procedure (MRPP). In MRPP, all possible paired combinations of either tributary types or mesohabitats were compared in multidimensional space to calculate standardized test statistics (T) and effect sizes (A) (McCune and Grace 2002, Peck 2010).

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Environmental analyses

The immediate environmental conditions experienced by macroinvertebrates were described using percentage cover of macrophytes, periphyton, filamentous algae, organics, embeddedness, and stream temperature, dissolved oxygen, pH, substrate particle size, depth, width, velocity, and discharge (Tables 10, 11). Percentage covers were measured within the slack-Surber area and all other remaining measures, excluding discharge, were collected directly upstream of the sampler. Linear correlations of environmental variables to macroinvertebrate community densities of all taxa were performed using canonical correspondence analysis (CCA) (PC-ORD, version 6). Plot validity was assessed using a randomization test where significance indicates the plot configuration is greater than that produced by chance. The strongest correlates were described using joint biplots of taxa and environmental variable vectors (McCune and

Grace 2002, Peck 2010). Statistically significant differences between tributaries and among mesohabitats were tested using a two-way ANOVA ( = 0.05) (SAS, version 9.4).

Homogeneity of variance was checked using residual variance plots and normality was tested using Shapiro-Wilks test. Where data did not meet ANOVA assumptions, data were either square root or log transformed. Pairwise comparisons were performed using t-tests with Bonferroni adjustments.

Results

Macroinvertebrate community descriptions

Mesohabitat comparisons were based on 20,826 individuals that included 144 taxa. Surface-fed tributary marginal vegetation harbored Epiaeshna, Hetaerina

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(Odonata), Allocapnia, Neoperla falayah Stark and Lentz (Plecoptera), Acentrella,

Hexagenia (Ephemeroptera), Ranatra, Rhagovelia, Saldidae (Hemiptera), Sialis

(Megaloptera), Oxyethira (Trichoptera), Gonielmis, Psephenus, Stenelmis (Coleoptera),

Hexatoma, Odontomyia, Probezzia, and Stratiomys (Diptera) (Appendix G).

Spring-fed tributaries were characterized by higher densities of Platyhelminthes,

Oligochaeta, Amnicola (Gastropoda), Asellus (Isopoda), Gammarus, Hyallela

(Amphipoda), Chelifera, and Chironomidae (Diptera) than in surface-fed tributaries.

Surface-fed tributaries were characterized by Gomphidae (Odonata), Leuctra

(Plecoptera), Caenis, Maccaffertium (Ephemeroptera), Cheumatopsyche (Trichoptera),

Hexatoma, Hemerodromia, and Simulium (Diptera). Other taxa collected only in marginal vegetation in the spring-fed tributaries included Asellus, Lirceus (Isopoda),

Gomphus (Odonata), Acerpenna (Ephemeroptera), Microvelia, Corixidae (Hemiptera),

Micrasema, Psychomyia, Triaenodes (Trichoptera), Berosus, Cymbiodytes, Enochrus,

Hydrobiomorpha, Hydrobius, Peltodytes (Coleoptera), Alluaudomyia, Cryptolabis,

Chrysops, Dixa, Gymnopais, Axymyiidae, Psychodidae, and Sciomyzidae (Diptera)

(Appendix H). Among all mesohabitats, marginal vegetation was the most variable taxonomically whereas pools were least variable. Marginal vegetation mesohabitats of both tributaries accounted for higher richness in Odonata, Ephemeroptera, Hemiptera,

Coleoptera, and Diptera.

Macroinvertebrate metric analyses

Among mesohabitats, riffles had high macroinvertebrate densities while pools had low densities although statistically significant differences were not detected (Table 12).

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Taxonomic richness was high in marginal vegetation (surface-fed = 80 taxa, spring-fed =

82 taxa) and low in pools (surface-fed = 59 taxa, spring-fed = 64 taxa). Metrics of dominance, Shannon’s Diversity Index, and taxonomic richness were significantly different among either tributaries or mesohabitats (Table 12). Dominance by

Chironomidae was greater in spring-fed tributaries than in surface-fed tributaries (p =

0.026) (Fig. 37). Shannon’s Diversity Index was highest in riffle and marginal vegetation mesohabitats (p = 0.017) (Fig. 38). Taxonomic richness was greater in surface-fed tributaries than in spring-fed tributaries (p = 0.005) (Figs. 39, 40). Taxonomic similarity between tributaries was most evident between surface-fed and spring-fed pools (J = 0.48)

(Table 13). The lowest similarity value occurred between the marginal vegetation of surface-fed and spring-fed tributaries (J = 0.36).

Taxonomic richness within functional feeding groups differed between tributary types (p = 0.001) (Fig. 41) and among mesohabitats (p = 0.011) (Fig. 42) (Table 14) in nonparametric tests. Collector, filterer, and scraper richness differences were also supported in parametric tests (Table 15). Collector richness was higher in surface-fed tributaries than in spring-fed tributaries (p = 0.018) (Fig. 43). Filterers (p = 0.013) (Fig.

44) and scrapers (p = 0.012) (Fig. 45) were richer in riffle mesohabitats than in pool mesohabitats. Densities within functional feeding groups between tributaries (Fig. 46) or among mesohabitats (Fig. 47) did not differ statistically (Table 16).

Macroinvertebrate community analyses

Taxonomic composition of macroinvertebrate communities in surface-fed

(Appendix G) and spring-fed tributaries (Appendix H) overlapped greatly in two-

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dimensional NMS space (Fig. 48), but were statistically different in composition (p <

0.001) (Table 14). Further, macroinvertebrate community composition significantly differed among mesohabitats (p < 0.001) (Table 14) (Fig. 49). Specifically, surface-fed mesohabitats were significantly different from each other and from spring-fed mesohabitats. Mesohabitats in spring-fed tributaries did not differ from one another.

Environmental analyses

Water temperature, pH, and discharge were the variables most strongly correlated with macroinvertebrate communities from mesohabitats (Fig. 50). Lowest temperatures were from spring-fed tributary riffles (mean = 15.05°C) and the highest temperatures were from surface-fed tributary marginal vegetation (22.62°C). Significant differences in parametric tests supported these nonparametric relationships. Temperature (p < 0.001)

(Fig. 51) and pH (p < 0.001) (Fig. 52) were higher in surface-fed tributaries than in spring-fed tributaries. Discharge was greater in spring-fed tributaries than in surface-fed tributaries (p = 0.005) (Fig. 53). Mesohabitat environments were characterized by differences in depth, percent embeddedness cover, percent macrophyte cover, percent organic cover, substrate particle size, and velocity (Table 17). Spring-fed riffles and surface-fed pools had the deepest waters (p = 0.011) (Fig. 54). Pool mesohabitats were the most embedded (p < 0.001) (Fig. 55). Macrophyte cover was highest in marginal vegetation mesohabitats (p < 0.001) (Fig. 56). Organic materials were highest in pool mesohabitats (p < 0.001) (Fig. 57). Substrate particle sizes were largest in riffle mesohabitats and lowest in pools (p < 0.001) (Fig. 58). Riffle mesohabitats had the fastest velocities (p = 0.004) (Fig. 59). Environmental characteristics were wide ranging

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across tributaries (Fig. 60), but these variables were still distinguishable in multidimensional space within like mesohabitats across tributary types (Fig. 61).

Discussion

Macroinvertebrate composition, diversity, and FFGs significantly differed between tributaries but not consistently among mesohabitats. These results suggest the species pools differ between surface-fed and spring-fed tributaries. The differences in species pools of tributaries can be attributed to dispersal constraints (see Parkyn and

Smith 2011). Constraints likely exist because both in-stream and aerial dispersal are restricted in low-order streams of a network, which reinforces isolation among tributaries in a watershed (Clarke et al. 2008). Distances between tributaries in a network grow as the order of the stream decreases. For example, tributaries in the Current and Jacks Fork rivers are separated by a minimum of two miles and a maximum of 27 miles. Most macroinvertebrates capable of flight are not likely to cross this distance because it exceeds known dispersal maximum of 0.5 miles for Plecoptera (MacNeale et al. 2005).

Also, higher macroinvertebrate community interaction and mixing is expected within mesohabitats, but less so at larger scales (Costa and Melo 2008). Dispersal constraints were supported in this study by higher similarities within replicates of a single mesohabitat than among replicates of multiple mesohabitats. Similar results to this study exist where 42% of macroinvertebrate community variation was explained using mesohabitats compared to only 22% explained by tributaries in streams of the Brazilian

Atlantic rain forest (Costa and Melo 2008).

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Disturbance regimes also can increase any differences among species pools.

Higher community interaction is expected within a tributary rather than among tributaries when disturbance regimes exceed colonization rates or where food resources are both plentiful and uniformly distributed (Resh et al. 1988). For example, while spring-fed tributaries of the Current and Jacks Fork rivers had an abundance of filamentous algae and macrophytes, surface-fed tributaries had denuded substrates lacking macrophytes.

However, abundant food resources in spring-fed tributaries suggests greater separation of macroinvertebrate communities among mesohabitats than would be found in surface-fed mesohabitats, although this was not revealed by community composition or Jaccard’s similarity scores. When food resources are abundant communities have less need to move to locate new food resources. The inability to detect greater disparity among macroinvertebrate communities according to a mesohabitat could also result from weather during sample collections. Sampling in the Current and Jacks Fork river tributaries occurred during several rain events which may have actually increased macroinvertebrate interactions (drift or migration into interstitial spaces) in tributaries, but this was not supported with significant differences found in composition and taxonomic richness across mesohabitats. In contrast, surface-fed tributaries had high within-mesohabitat variation in community structure suggesting different levels of disturbance were acting on replicates in this study.

Surface-fed and spring-fed tributaries should support distinct macroinvertebrate communities due to contrasting baseline environmental attributes. Unique spring macroinvertebrate faunas exist due to characteristically stable flow and temperatures that enable increased biotic interactions. Therefore, macroinvertebrate colonization tends to

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be limited by the biota rather than by stochastic processes (Glazier 1991, Cantonati et al.

2012). As a result, springs should favor taxa capable of competition rather than survival during flood or drought (Kubikova et al. 2012). Furthermore, stable flows in spring-fed tributaries buffer against rainfall events (Brooks 2009). Spring-fed tributary replicates were closer together on NMS plots, which indicated effects from the rain were buffered to some degree. Competition rather than disturbance was likely the reason for greater within-tributary macroinvertebrate community interactions. Previous work indicated that aggressive competition by the invasive snail, Potamopyrgus antipodarum (Gray), excluded other macroinvertebrates from areas where it grazed (Kerans et al. 2005).

Although competition was unmeasured in spring-fed tributaries of the Current and Jacks

Fork rivers, snails composed 16% of the samples compared to only 3% in surface-fed tributaries, which indicates competition could have been heavier in springs. Other biotic pressures come from predators that usually are twice as abundant in spring-fed tributaries as in surface-fed tributaries and have the ability to reduce total diversity in stable spring systems (Barquin and Death 2004). However, in this Ozark system, predation pressures were not high in spring-fed tributaries as measured by the predator functional feeding group. In contrast to the biotic pressures of spring-fed tributaries, surface-fed tributaries undergo more water level fluctuations over the course of a year. As a result, mesohabitats repeatedly connect and disconnect depending upon rain (Lisle 1982).

Therefore, surface-fed communities should favor taxa able to efficiently disperse rather than compete. Spring-fed tributary macroinvertebrate communities of this study were buffered by biotic pressures while surface-fed tributary communities were more influenced by stochastic events.

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Functional feeding groups are a reflection of food resource availability and support community differences between tributaries and among mesohabitats. Despite this study’s focus on mesohabitats with different food resources, macroinvertebrate communities of the entire spring-fed tributary were characterized by abundant shredders.

Shredders consume the course particulate organic matter (CPOM) and macrophytes

(Cummins et al. 2008), which were abundant in all spring-fed tributary replicates.

Springs also favor primary producers such as filamentous algae and bryophytes more so than do surface-fed tributaries (Barquin and Death 2006), which was documented in the tributaries of the Current and Jacks Fork rivers. Although surface-fed tributaries have lower primary productivity than do spring-fed tributaries, they have an abundance of detritus from leaf litter and mechanical decomposition by larger vertebrates (Clarke et al.

2008). In surface-fed tributaries, collector-gatherer densities were high, suggesting abundant fine particulate organic matter (FPOM) (Cummins et al. 2008). Interestingly, habitat stability can be determined by such high percentages of collector-gatherers

(Sullivan et al. 2004). This contradicts assumptions concerning habitat stability in spring-fed tributaries because particle sizes in streams studied here were large and should support greater habitat stability. Disturbance from rain events might have affected FFG interpretations or the coarse groupings formed from FFGs may be poor indications of environmental conditions in the tributaries of this study.

Macroinvertebrate studies incorporating multiple mesohabitats are valuable to taxonomists, managers, and ecologists. They help aquatic ecologists understand macroinvertebrate distributions, dispersal, and interactions. Taxonomically, multiple mesohabitats provide researchers with a stronger foundation to understand the regional

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species pools. In this study, uncommon taxa in riffles were frequently recorded and these can become useful in the documentation of habitat changes over time. In small-scale studies, the most cost-effective sampling regimes include five mesohabitats, whereas large-scale studies needed to sample only pools (Chessman et al. 2007). In this way, studies using multiple mesohabitats are necessary in order to explore best regional protocols.

Ecologically, multimesohabitat studies further the exploration of stream communities in order to better investigate the effects of anthropogenic pressures.

Climate change studies emphasize the need for more sensitive or specialized metrics to measure changes in freshwaters unrelated to severe physical disturbances. In these metric revisions, studies are including fine-resolution taxonomy combined with experimentation of understudied groups such as Chironomidae (Wymer and Cook 2003).

Based on the Wymer and Cook study, Chironomidae were taxonomically richer in pools and other slack-flow mesohabitats than in riffles. Therefore, slack-flow habitats could be important for inclusion in climate change protocols. Overall, this study suggests environmental parameters alone should be used in conjunction with macroinvertebrate samples to best interpret habitat conditions.

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Table 10. Environmental variables taken in July and September 2013 before Brown vacuum samples were collected in mesohabitats from surface-fed tributaries of the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation, D.O. = Dissolved oxygen, Wentwth. = Wentworth. Values are based on means from three subsamples.

Ashley Creek Bay Creek Big Creek Rocky Creek Sinking Creek Variable Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg D. O. (mg L-1) 7.5 7.5 7.9 11.1 9.3 9.5 999.0 4.5 1.5 7.4 5.3 5.4 7.4 4.9 8.6 pH 8.0 7.9 8.0 7.8 7.8 7.8 7.5 7.6 7.7 7.8 7.9 8.0 7.5 8.0 7.7 Temperature (°C) 24.3 24.3 24.4 21.4 21.3 21.4 20.7 22.0 21.9 22.7 22.9 23.7 21.5 21.6 21.7 Embeddedness (%) 25.0 77.5 56.7 25.0 46.7 25.0 46.7 77.5 35.8 57.5 87.5 35.8 25.0 67.5 25.0 Filamentous Algae (%) 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 1.7 0.0 0.0 0.0 19.2 10.0 Macrophytes (%) 0.0 0.0 56.7 0.0 0.0 45.8 0.0 0.0 77.5 0.0 0.0 67.5 0.0 8.3 77.5 Organics (%) 8.3 56.7 5.0 18.3 87.5 29.2 22.5 46.7 18.3 18.3 25.0 57.5 5.0 77.5 25.0 Periphyton (%) 18.3 16.7 18.3 18.3 11.7 25.0 57.5 35.8 18.3 57.5 57.5 57.5 25.0 19.2 25.0

Substrate Size (Wentwth.) 12.6 6.1 7.9 13.2 11.6 11.6 13.7 7.6 8.8 12.8 8.9 10.2 12.0 7.2 11.7 108

Depth (m) 0.22 0.16 0.06 0.11 0.26 0.13 0.13 0.20 0.10 0.29 0.41 0.14 0.11 0.36 0.08

Discharge (m3 s-1) 0.4 0.4 0.4 0.1 0.1 0.1 0.2 1.4 1.4 0.1 0.1 0.1 1.8 1.8 1.8 Velocity (m s-1) 0.6 0.1 0.1 0.4 0.0 0.0 0.6 0.1 0.1 0.3 0.0 0.0 0.3 0.1 0.3 Width (m) 8.8 8.8 8.8 6.1 6.1 6.1 18.0 19.3 19.3 17.0 17.0 17.0 27.2 27.2 27.2

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Table 11. Environmental variables taken in July and September 2013 before Brown vacuum samples were collected in mesohabitats from spring-fed tributaries of the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation, D. O. = Dissolved oxygen, Wentwth. = Wentworth. Values are based on means from three subsamples. Values are based on means from three subsamples.

Alley Spring Blue Spring Cave Spring Pulltite Spring Round Spring Variable Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg D. O. (mg L-1) 5.9 5.5 5.5 13.1 13.2 9.7 4.7 5.6 4.4 6.3 6.3 6.4 7.8 6.8 7.3 pH 7.4 7.3 7.2 6.8 7.1 7.2 7.1 7.6 7.7 7.4 7.3 7.4 7.4 7.3 7.3 Temperature (°C) 14.4 14.6 14.3 13.9 14.5 14.0 18.0 18.5 18.2 14.0 14.0 14.0 15.0 14.6 19.1 Embeddedness (%) 25.0 77.5 35.8 25.0 56.7 35.8 46.7 66.7 57.5 25.0 77.5 25.0 25.0 87.5 35.8 Filamentous Algae (%) 1.7 0.0 8.3 8.3 1.7 20.8 20.8 0.0 0.0 0.0 0.0 0.0 3.3 0.0 37.5 Macrophytes (%) 29.2 0.0 58.3 0.0 3.3 87.5 8.3 0.0 57.5 10.0 0.0 77.5 0.0 0.0 35.8 Organics (%) 5.0 87.5 46.7 5.0 29.2 5.0 29.2 67.5 35.8 5.0 56.7 5.0 11.7 50.0 18.3 Periphyton (%) 18.3 38.3 18.3 5.0 5.0 5.0 35.8 46.7 46.7 5.0 11.7 5.0 11.7 37.5 29.2

109 Substrate Size (Wentwth.) 13.0 3.6 10.7 13.1 7.3 7.4 11.4 9.5 9.0 13.6 3.8 12.0 12.7 1.7 8.7

Depth (m) 0.42 0.28 0.17 0.28 0.14 0.18 0.21 0.25 0.11 0.29 0.12 0.11 0.53 0.20 0.16 Discharge (m3 s-1) 0.9 0.9 0.9 2.3 2.3 2.3 0.7 0.7 0.7 1.7 1.7 1.7 1.3 1.3 1.3 Velocity (m s-1) 0.3 0.0 0.2 0.8 0.1 0.3 0.4 0.0 0.1 0.9 0.0 0.1 0.3 0.1 0.0 Width (m) 6.5 6.5 6.5 20.9 20.9 20.9 9.9 9.9 9.9 8.5 8.5 8.5 13.4 13.4 13.4

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Table 12. Two-way analyses of variance comparing metrics between tributaries and among mesohabitats in July and September 2013 from the Current and Jacks Fork rivers, Missouri.

Variable Effect NDF DDF F p Density Tributary 1 24 0.05 0.829 Mesohabitats 2 24 1.99 0.159 Tributary x Mesohabitat 2 24 0.28 0.761 Dominance Tributary 1 24 5.65 0.026 * Mesohabitats 2 24 2.31 0.121 Tributary x Mesohabitat 2 24 0.20 0.823 Shannon's Diversity Tributary 1 24 1.44 0.241 Index Mesohabitats 2 24 4.85 0.017 * Tributary x Mesohabitat 2 24 0.26 0.776 Taxonomic Richness Tributary 1 24 9.55 0.005 ** Mesohabitats 2 24 1.90 0.172 Tributary x Mesohabitat 2 24 0.45 0.645 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

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Table 13. Jaccard’s similarity scores (J) for pairwise comparisons between tributaries and mesohabitats of the Current and Jacks Fork rivers, Missouri in July and September 2013.

Comparison J Surface Tributary v. Spring Tributary 0.58 Surface Riffle v. Surface Pool 0.43 Surface Pool v. Surface Vegetation 0.49 Surface Riffle v. Surface Vegetation 0.52 Spring Riffle v. Spring Pool 0.49 Spring Pool v. Spring Vegetation 0.51 Spring Riffle v. Spring Vegetation 0.45 Surface Riffle v. Spring Riffle 0.44 Surface Pool v. Spring Pool 0.48 Surface Vegetation v. Spring Vegetation 0.36

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Table 14. Multi-response permutation procedure tests associated with nonmetric multidimensional scaling between tributaries and among mesohabitats in the Current and Jacks Fork rivers, Missouri in July and September 2013. Community refers to macroinvertebrate community. R = riffle, P = pool, V – marginal vegetation. FFG = functional feeding group.

Variable Source T A p Community Tributary -7.822 0.049 <0.001 *** Mesohabitat -4.089 0.062 <0.001 *** Surface R v. Spring P -3.983 0.072 0.002 ** Surface R v. Spring V -4.538 0.073 0.001 ** Surface P v. Spring R -3.626 0.090 0.003 ** Surface P v. Spring P -2.159 0.052 0.033 * Surface P v. Spring V -3.808 0.081 0.003 ** Surface V v. Spring R -1.951 0.046 0.043 * Surface V v. Spring P -2.137 0.043 0.033 * Surface V v. Spring V -2.709 0.052 0.014 * Densities within FFGs Tributary -0.532 0.009 0.218 Mesohabitat -0.132 0.005 0.403 Taxonomic Tributary -6.024 0.102 0.001 *** richness within FFGs Mesohabitat -2.824 0.116 0.011 * Surface R v. Surface P -2.655 0.122 0.017 * Surface R v. Surface V -2.186 0.091 0.029 * Surface R v. Spring R -2.113 0.165 0.043 * Surface R v. Spring P -3.721 0.192 0.006 ** Surface P v. Spring V -3.022 0.162 0.013 * Surface V v. Spring P -2.496 0.131 0.026 * * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

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Table 15. Two-way analyses of variance of functional feeding group (FFG) richness between tributaries and among mesohabitats of the Current and Jacks Fork rivers, Missouri in July and September 2013.

Variable Effect NDF DDF F p Collector Tributary 1 24 6.42 0.018 * Mesohabitat 2 24 0.98 0.389 Tributary x Mesohabitat 2 24 0.13 0.877 Filterer Tributary 1 24 2.10 0.160 Mesohabitat 2 24 5.23 0.013 * Tributary x Mesohabitat 2 24 3.36 0.052 Predator Tributary 1 24 1.93 0.178 Mesohabitat 2 24 1.19 0.322 Tributary x Mesohabitat 2 24 0.49 0.621 Scraper Tributary 1 24 20.11 <0.001 *** Mesohabitat 2 24 5.34 0.012 * Tributary x Mesohabitat 2 24 0.35 0.711 Shredder Tributary 1 24 0.42 0.524 Mesohabitat 2 24 1.81 0.185 Tributary x Mesohabitat 2 24 0.14 0.871 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

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Table 16. Two-way analyses of variance of functional feeding group (FFG) densities between tributaries and among mesohabitats of the Current and Jacks Fork rivers, Missouri in July and September 2013.

Variable Effect NDF DDF F p Collector Tributary 1 24 1.39 0.250 Mesohabitat 2 24 0.46 0.638 Tributary x Mesohabitat 2 24 0.06 0.944 Filterer Tributary 1 24 0.17 0.685 Mesohabitat 2 24 4.97 0.016 Tributary x Mesohabitat 2 24 1.16 0.331 Predator Tributary 1 24 0.98 0.333 Mesohabitat 2 24 0.94 0.404 Tributary x Mesohabitat 2 24 0.07 0.935 Scraper Tributary 1 24 0.14 0.711 Mesohabitat 2 24 2.90 0.075 Tributary x Mesohabitat 2 24 0.56 0.579 Shredder Tributary 1 24 0.08 0.782 Mesohabitat 2 24 1.01 0.380 Tributary x Mesohabitat 2 24 1.14 0.335 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

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Table 17. Two-way analyses of variance of environmental variables between tributaries and among mesohabitats in the Current and Jacks Fork rivers, Missouri in July and September 2013. Dashes indicate data did not meet assumptions of ANOVA after transformation.

Variable Effect NDF DDF F p Depth Tributary 1 24 3.62 0.069 Mesohabitat 2 24 10.49 <0.001 *** Tributary x Mesohabitat 2 24 5.49 0.011 * Discharge Tributary 1 24 9.55 0.005 ** Mesohabitat 2 24 0.15 0.865 Tributary x Mesohabitat 2 24 0.15 0.865 Dissolved oxygen Tributary - - - - Mesohabitat - - - - Tributary x Mesohabitat - - - - Embeddedness Tributary 1 24 0.03 0.871 Mesohabitat 2 24 28.01 <0.001 *** Tributary x Mesohabitat 2 24 0.36 0.699 Filamentous algae Tributary - - - - Mesohabitat - - - - Tributary x Mesohabitat - - - - Macrophytes Tributary 1 24 1.84 0.188 Mesohabitat 2 24 101.52 <0.001 Tributary x Mesohabitat 2 24 3.02 0.068 Organics Tributary 1 24 0.74 0.399 Mesohabitat 2 24 12.77 <0.001 *** Tributary x Mesohabitat 2 24 0.22 0.805 Periphyton Tributary 1 24 2.96 0.098 Mesohabitat 2 24 0.12 0.891 Tributary x Mesohabitat 2 24 0.82 0.454 pH Tributary 1 24 48.61 <0.001 *** Mesohabitat 2 24 1.23 0.311 Tributary x Mesohabitat 2 24 3.62 0.954 Substrate Tributary 1 24 3.05 0.093 Mesohabitat 2 24 25.17 <0.001 *** Tributary x Mesohabitat 2 24 1.85 0.179 Temperature Tributary 1 24 110.95 <0.001 *** Mesohabitat 2 24 0.39 0.684 Tributary x Mesohabitat 2 24 0.09 0.916 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001 115

Table 17. Continued.

Variable Effect NDF DDF F p Velocity Tributary 1 24 0.98 0.332 Mesohabitat 2 24 7.16 0.004 ** Tributary x Mesohabitat 2 24 0.39 0.679 Width Tributary 1 24 2.05 0.165 Mesohabitat 2 24 0.00 0.999 Tributary x Mesohabitat 2 24 0.00 0.999 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

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Big Creek Ashley Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Bay Creek

Flow Cave Spring Alley Spring Rocky Creek

Figure 35. Macroinvertebrate sampling tributary sites of July and September 2013 in surface-fed tributary sites (Ashley Creek, Bay Creek, Big Creek, Rocky Creek, Sinking Creek) and spring-fed tributary sites (Alley Spring, Blue Spring, Cave Spring, Pulltite Spring, Round Spring). Tributaries are associated with the Current and Jacks Fork rivers, Missouri, which are shown in bold.

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Mesh collection Water Inflow bag inside water intake chamber Water Outflow

Figure 36. Brown vacuum sampler used to collect macroinvertebrates in July 2013 from the Current and Jacks Fork rivers, Missouri.

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0.95 a

0.9 b

0.85

0.8

0.75 Dominance

0.7

0.65 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats

Figure 37. Two-way analysis of variance comparing dominance among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

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1.2 a a

1 b

0.8

0.6

0.4

Shannon’s Diversity Index Diversity Shannon’s 0.2

0 Surface R Spring R Surface P Spring P Surface V Spring V Mesohabitats

Figure 38. Two-way analysis of variance comparing Shannon’s Diversity Index among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

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50 a

45 b 40

35

30

25

20

15 Taxonomic Richness Taxonomic 10

5

0 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats

Figure 39. Two-way analysis of variance comparing taxonomic richness among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

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90 80 Diptera Coleoptera 70 Trichoptera 60 Megaloptera Hemiptera Richness 50 Plecoptera 40 Ephemeroptera 30 Odonata 20 Amphipoda Taxonomic Taxonomic Isopoda 10 Pelecypoda 0 Gastropoda Riffle Pool Veg Riffle Pool Veg Surface Spring

Figure 40. Composite graph of order-level richness among mesohabitats from July 2013 in the Current and Jacks Fork rivers, Missouri. Surface = surface-fed, Spring = spring- fed, Veg = marginal vegetation.

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NMS 2 NMS

Tributary Types Surface-fed Spring-fed + Centroid NMS 1

Figure 41. Nonmetric multidimensional scaling plot for July 2013 Brown vacuum samples showing functional feeding group richness from the Current and Jacks Fork rivers, Missouri. Stress = 7.47, randomization p = 0.004.

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Mesohabitats Surface

NMS2 Riffles Veg Pools

Riffles Spring Veg Pools + Centroid NMS 1

Figure 42. Nonmetric multidimensional scaling plot for July 2013 Brown vacuum samples showing functional feeding group richness among mesohabitats from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, Spring = spring-fed tributaries, Veg = marginal vegetation. Stress = 7.47, randomization test p = 0.004.

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20 a

18 b

16

14

12

10

8

Collector Richness Collector 6

4

2

0 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats

Figure 43. Two-way analysis of variance comparing collector richness among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

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6 a

5

4 ab

3 b

2 Filterer Richness Filterer

1

0 Surface R Spring R Surface P Spring P Surface V Spring V Mesohabitats

Figure 44. Two-way analysis of variance comparing filterer richness among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

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a 16

14 b b 12

10

8

6

4 Scraper RichnessScraper 2

0 Surface R Spring R Surface P Spring P Surface V Spring V Mesohabitats

Figure 45. Two-way analysis of variance comparing scraper richness among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

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NMS 2 NMS

Tributary Types Surface-fed Spring-fed + Centroid

NMS 1

Figure 46. Nonmetric multidimensional scaling plot for July 2013 Brown vacuum samples showing functional feeding group densities from the Current and Jacks Fork rivers, Missouri. Stress = 10.85, randomization p = 0.004.

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Mesohabitats

Riffles Surface NMS 2 NMS Veg Pools

Riffles Spring Veg Pools + Centroid NMS 1

Figure 47. Non-metric multidimensional scaling plot for July 2013 Brown vacuum samples showing functional feeding group densities among mesohabitats from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, Spring = spring-fed tributaries, Veg = marginal vegetation, Stress = 10.85, randomization p = 0.004.

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NMS 2 NMS

Tributary Types Surface-fed Spring-fed + Centroid NMS 1

Figure 48. Non-metric multidimensional scaling plot for July 2013 Brown vacuum samples showing macroinvertebrate communities from the Current and Jacks Fork rivers, Missouri. Stress = 14.3, randomization p = 0.004.

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Mesohabitats

Riffles Surface NMS2 Veg Pools

Riffles Spring Veg Pools + Centroid NMS 1

Figure 49. Non-metric multidimensional scaling plot for July 2013 Brown vacuum samples showing macroinvertebrate communities from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, Spring = spring-fed tributaries, Veg = marginal vegetation, Stress = 14.3, randomization p = 0.004.

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CCA 2 CCA

CCA 1

Figure 50. Canonical correspondence analysis biplot for July 2013 Brown vacuum samples showing macroinvertebrate community correlations to environmental variables. Arrow length indicates strength of correlation. Arrow direction indicates direction of relationship between variable and community. R = riffle, P = pool, V = marginal vegetation.

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25 a

20

b

C)

15

10 Temperature ( Temperature 5

0 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats Figure 51. Two-way analysis of variance comparing temperature among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

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8 a

7.8

7.6 b

7.4 pH

7.2

7

6.8

6.6 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats

Figure 52. Two-way analysis of variance comparing pH among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

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1.8 a

1.6

) 1.4

1 -

s b

3 1.2

1

0.8

0.6 Discharge (m Discharge 0.4

0.2

0 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats

Figure 53. Two-way analysis of variance comparing discharge among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences based on a square root transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

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45 a 40

35 a

30 b 25 b 20 bc

Depth (cm) Depth 15 c 10

5

0 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats

Figure 54. Two-way analysis of variance comparing depth among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences are based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

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90 a 80

70 (%) 60

50 b b

40

30

20 Embeddedness 10

0 Surface R Spring R Surface P Spring P Surface V Spring V Mesohabitats

Figure 55. Two-way analysis of variance comparing embeddedness among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

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80 a

70

60

(%) 50

40

30

20 Macrophytes b b 10

0 Surface R Spring R Surface P Spring P Surface V Spring V -10 Mesohabitats

Figure 56. Two-way analysis of variance comparing macrophyte cover among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences based on a square root transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

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80 a

70

60

50

40 b

30 Organics (%) Organics b 20

10

0 Surface R Spring R Surface P Spring P Surface V Spring V Mesohabitats

Figure 57. Two-way analysis of variance comparing organics among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences are based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

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14 a

12 b

10 c

8

6 (Wentworth)

4 Substrate particle size size particle Substrate 2

0 Surface R Spring R Surface P Spring P Surface V Spring V Mesohabitats

Figure 58. Two-way analysis of variance comparing substrate particle sizes among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

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0.8 a

0.7

) 1 - 0.6

0.5

0.4

0.3 Velocity (m s (m Velocity b 0.2

0.1 b

0 Surface R Spring R Surface P Spring P Surface V Spring V Mesohabitats

Figure 59. Two-way analysis of variance comparing velocity among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences based on a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

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NMS 2 NMS

Tributary Types Surface-fed Spring-fed + Centroid NMS 1

Figure 60. Non-metric multidimensional scaling plot for July 2013 Brown vacuum samples showing environmental variables between tributary types in the Current and Jacks Fork rivers, Missouri. Stress = 15.36, randomization p = 0.004.

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Mesohabitats

Riffles Surface

NMS 2 NMS Veg Pools

Riffles Spring Veg Pools + Centroid NMS 1

Figure 61. Non-metric multidimensional scaling plot for July 2013 Brown vacuum samples showing environmental variables among mesohabitats in the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, Spring = spring-fed tributaries, Veg = marginal vegetation. Stress = 15.36, randomization p = 0.004.

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CHAPTER 5

Chironomidae in the Ozarks and the Importance of Taxonomic Resolution

Abstract

Chironomidae are important in stream benthic studies because their abundance and diversity were previously documented to be as much as 42% of Ozark stream communities with as many as 25 taxa. However, the taxonomic resolution at the generic or specific levels is poorly understood, which could impede ecological interpretations of the environment. The inclusion of Chironomidae in bioassessments could lead to earlier and more sensitive detection of disturbance. The objectives of this study were to determine whether or not a regional bioassessment would be altered by the inclusion of chironomid genera and to describe and quantify the chironomid communities from riffles, pools, and marginal vegetation in surface-fed and spring-fed tributaries of the Current and Jacks Fork rivers, Missouri. In the summer of 2013, 10 tributaries were sampled using a Brown vacuum sampler. From 48 genera and 840 individuals, interpretation of the SCI was unaltered by chironomid genera inclusion. The only metric affected by chironomid inclusion was taxonomic richness. Taxonomic richness was higher in surface-fed tributaries than in spring-fed tributaries. Diversity and the SCI were also greater in surface-fed tributaries than in spring-fed tributaries. Biotic index values were higher in pool than riffle mesohabitats. Inclusion of chironomid genera in NMS plots did not alter the interpretation of intercommunity relationships among mesohabitats or

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between tributary types. The greatest overlap in chironomid communities across tributary types was in pools. Interestingly, genera of chironomids alone in NMS space could also distinguish among mesohabitats and subfamily densities of chironomids differed among mesohabitats. Important variables for the most abundant chironomids included fine sediments and nutrient levels. These results indicate the organization of chironomid communities of the Current and Jacks Fork rivers are consistent with our current understanding of general macroinvertebrate community structure and relationships to the environment. The potential exists to interpret habitat degradation due to chironomid association with sediments and nutrients, which commonly are a reflection of bank stability and land use issues in watersheds.

Introduction

Chironomidae compose a major portion of the Diptera community in freshwater habitats (Pinder 1986, Lencioni and Rossaro 2005) and contribute largely to the vast array of benthic macroinvertebrates. Missouri Ozark Chironomidae represent 32–42% of the total benthos and includes 25 genera in reference quality streams (Rabeni and Wang

2001). These abundant larvae and pupae also provide an important food resource for other macroinvertebrates and vertebrates such as fish. Critical trophic links between primary producers and fish are made by chironomids, especially during early fish development (Berg and Hellenthal 1992, Rabeni and Wang 2001).

The taxonomic resolution of Chironomidae is improving with the help of regional keys. For example, a key to the southeastern U.S. chironomids was given by Epler

(2001), the southwestern U.S. Chironomidae were studied intensively by Wiederholm

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(1983), and a summary of available keys to the genus level was provided by Ferrington et al. (2008) for North America. Despite growing taxonomic efforts in the Chironomidae, the ecology and life cycles for most species are still poorly understood because of low accessibility and understanding of taxonomic keys and the vast number of species (Pinder

1986). Complete stream habitat interpretation and ecological trends rely fundamentally upon the need for finer taxonomic resolution despite difficulties in preparing specimens for study (Pond et al. 2008).

The use of Chironomidae in water quality assessment is debatable and depends upon the metrics chosen. Their use in bioassessments is controversial due principally to the amount of effort required for identifications. In a single sample, the time to identify chironomids is equal to the time it takes to identify 50% to 100% of the entire sample

(Rabeni and Wang, 2001). Sometimes, chironomid inclusion in bioassessments does not change community or environmental interpretations (Wright et al. 1995, Hewlett 2000).

Poor assessment performance occurs, in part, because of buffering effects from chironomid genera, which boost richness metrics (Rabeni and Wang 2001).

Despite these ongoing debates, Chironomidae deserve further exploration in both bioassessment and ecological studies. Stream macroinvertebrate community identifications at the species level are commonly performed in Europe (Waite et al.

2004), but less so in North American simply due to the lack of experience with

Chironomidae. With continued biological and ecological studies of chironomid genera and species, accessibility and taxonomy will improve bioassessment interpretation and design. Further, detection of subtle disturbances is possible, which will improve prospects for conservation efforts (Greffard et al. 2011).

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Studies of Chironomidae using multivariate techniques can improve ecological and habitat assessment accuracy in comparison to univariate methods that mask the information available at finer taxonomic levels (Wymer and Cook 2003). Chironomid identification is a necessary investment in order to bridge the gap in ecological understanding of this largely overlooked family. Taxonomic investments today in

Chironomidae will improve scientific studies over time and space, which will be helpful in analyses of long-term and large-scale studies.

The goal of this study was to assess chironomid communities in Ozark streams and to determine whether increasing taxonomic resolution of Chironomidae to the genus level would alter interpretation of the Missouri-adapted bioassessment, referred to as the

Stream Condition Index. Specific objectives included assessing and quantifying the chironomid genera in riffles, pools, and marginal vegetation mesohabitats and comparing the chironomid communities in surface-fed and spring-fed tributary habitats.

Methods

Macroinvertebrates were sampled in July (September for some samples due to rain) 2013 from tributaries flowing into the Current or Jacks Fork rivers in Carter, Dent, and Shannon counties, Missouri. Based on dominant groundwater, tributaries were classified as either spring-fed (groundwater dominant) or surface-fed (non-groundwater dominant). Ten tributaries were selected based on the presence of three comparable mesohabitats including surface-fed [Ashley Creek, Bay Creek, Big Creek (West), Rocky

Creek, and Sinking Creek] and spring-fed (Alley Spring, Blue Spring, Cave Spring,

Pulltite Spring, and Round Spring) tributaries (Fig. 35). Comparisons between spring-fed

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and surface-fed tributaries were made using five replicates. Mesohabitats were selected to maximize differences in stream velocity and depth (Cluer and Thorne 2013). Riffles had the fastest stream velocities, shallowest depths, few macrophytes, and largest substrate sizes. Pools had slow to zero stream velocities, deepest water, fewest macrophytes, abundant detritus, and smallest substrate sizes. Marginal vegetation had slow to zero stream velocities, shallow water, highest macrophytes, and medium substrate sizes.

Chironomidae were collected from three mesohabitats within a tributary using a

Brown vacuum sampler (sample area = 0.05 m2) for a duration of five minutes while disturbing the substrate with a garden rake. Macroinvertebrates were collected using a

500 m mesh collection bag (Brown et al. 1987). Chironomidae for analysis were chosen by randomly selecting 1/8 of a Petri plate holding the sample. Chironomidae were preserved in 80% ethanol and later processed in the laboratory. Processed

Chironomidae were identified to the lowest practical level using keys in Epler (2001) and

Ferrington et al. (2008). Subsamples were selected by random distribution of

Chironomidae in a Petri plate where genus-level identifications were performed for 1/8 of the total sample. Chironomidae identifications were confirmed by Brandy Bergthold of the Missouri Department of Natural Resources.

Chironomidae in tributaries and mesohabitats were compared using a split-plot design two-way analysis of variance (ANOVA) of subfamily densities and taxonomic richness where tributary and mesohabitats were factors (SAS, version 9.4). Additionally, three-way ANOVAs were used to assess taxonomic richness (TR), Shannon’s Diversity

Index (H’), biotic index (BI), and Stream Condition Index (SCI) when Chironomidae

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where either included or excluded from analyses of tributaries and mesohabitats. The

SCI index combines four metrics: total taxonomic richness (TR); Ephemeroptera,

Plecoptera, and Trichoptera richness (EPT); biotic index (BI); and Shannon’s Diversity

Index (H’). Each of these metrics in the SCI is standardized to scores of 1, 3, or 5 and then these scores are summed (Bowles et al. 2007). Taxonomic richness of

Chironomidae was also considered separately using two-way ANOVA comparing between tributaries and among mesohabitats. Heterogeneity of variance was tested using residual variance plots and normality using Shapiro Wilks test. Where statistical significance existed in the model ( = 0.05), pairwise comparisons were made using t- tests with Bonferroni adjustments for multiple comparisons.

Macroinvertebrate community differences were assessed in multidimensional space using nonmetric multidimensional scaling (NMS) where tributaries and mesohabitats were expected to form unique clusters. Smaller distances between points in

NMS space indicate greater similarity among communities. Taxa-by-site matrices were used in NMS to find the lowest stress in two- or three-dimensional plots that maximize distance between tributaries and among mesohabitats. A Sorensen distance measure was used to configure plots in multidimensional space using either two or three axes depending on three autopilot trials (PC-ORD, version 6) (Peck 2010). Plots were the result of random starting coordinates and 1000 runs. Plot significance was determined by randomizing data and comparing new configuration divergence to original configurations with 250 runs and 500 iterations to find stress. Stresses lower than 15 reflected plots representative of true multidimensional distances in ecological data. Significance in taxonomic composition was tested by a multi-response permutation procedure (MRPP) in

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which tributary types or mesohabitats were used as grouping variables to calculate standardized test statistics (T) and effect sizes (A) across all possible pairwise combinations (McCune and Grace 2002, Peck 2010).

Multiple linear regression was used to determine environmental relationships with the densities of the 10 densest chironomids. A stepwise method was used with  = 0.05

(SPSS, version 4.0) with 13 environmental variables: percentage cover of macrophytes, periphyton, filamentous algae, organics, and embeddedness, and stream temperature, dissolved oxygen, pH, substrate size, depth, width, velocity, and discharge.

Results

The Chironomidae collected included 840 individuals from 48 genera and three subfamilies: Chironominae, Orthocladinae, and Tanypodinae (Table 18). Chironominae were most prevalent followed by the Tanypodinae and Orthocladinae (Appendices I, J).

Spring-fed tributaries harbored 29 genera and surface-fed tributaries 39 genera. Total

Chironomidae richness in spring-fed tributary riffles, pools, and marginal vegetation mesohabitats was 13, 23, and 18 respectively. In surface-fed tributaries, total

Chironomidae richness was 20, 28, and 27 in riffles, pools, and marginal vegetation mesohabitats, respectively. Overall, pool mesohabitats were the most taxonomically rich in chironomids in both tributary types.

The addition of chironomid genera in analyses had little effect on metrics. Only taxonomic richness was increased with the addition genera of Chironomidae in the analysis where inclusion of Chironomidae increased taxonomic richness (p < 0.001) (Fig.

62) (Table 19). The remaining differences were between tributaries or mesohabitats.

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Community taxonomic richness was higher in surface-fed tributaries than in spring-fed tributaries (p < 0.001) (Fig. 63). Taxonomic richness of Chironomidae was higher in surface-fed tributaries than in spring-fed tributaries (p = 0.027) (Fig. 64) (Table 20).

Shannon’s Diversity Index was higher in surface-fed tributaries than in spring-fed tributaries (p < 0.001) (Fig. 65) (Table 19). The biotic index differed among mesohabitats where pools had higher values than riffles (p = 0.033) (Fig. 66).

Interpretation of the Stream Condition Index was unaltered with the inclusion of chironomid genera in the analysis (p = 0.843), but the SCI did differ between tributaries having higher values in surface-fed tributaries than in spring-fed tributaries (p = 0.033)

(Fig. 67).

In multivariate space, the addition of genera of chironomids in analyses maintained the distinction between tributary types and among mesohabitats based on macroinvertebrate community composition. Without chironomids, tributary types were separated widely in multidimensional space (p = 0.002) (Fig. 68) (Table 21). The addition of chironomid genera maintained these same relationships (p = 0.002) (Fig. 69).

Further, mesohabitat communities had similar relationships in multivariate space when chironomid genera were excluded (p < 0.001) (Fig. 70) or included (p < 0.001) (Fig. 71).

Each surface-fed mesohabitat significantly differed from each spring-fed mesohabitat in analyses with or without Chironomidae (Table 21). When Chironomidae were considered separately, the genera from pool mesohabitats were most similar across tributary types and showed the most overlap in multidimensional space (Fig. 72). The chironomid community alone could still discern unique communities present among mesohabitats (p < 0.001) (Table 22).

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Chironomidae subfamily densities differed between tributaries and among mesohabitats (Table 22). Chironominae were densest in surface-fed tributary riffles than in remaining mesohabitats (p = 0.040) (Fig. 73). Predatory Tanypodinae were denser in surface-fed tributaries than in spring-fed tributaries (p = 0.007) (Fig. 74); and

Tanypodinae were densest in pool mesohabitats (p = 0.026) (Fig. 75). Orthocladinae were densest in riffle mesohabitats (p < 0.001) (Fig. 76).

Evaluation of environmental relationships to chironomids was supported by using the ten most densely occurring genera which revealed a tendency for associations with fine sediments and higher nutrients. The densest taxa of Chironomidae included

Polypedilum, Thienemannimyia group, Cricotopus/Orthocladius group, Tanytarsus,

Ablabesmyia, Cladotanytarsus, Rheotanytarsus, Rheocricotopus, Parametriocnemus, and

Nilotanypus (Fig. 77) where Polypedilum accounted for 22% of all individuals in samples. Significant linear relationships were detected for six of the ten genera and habitat variables were related largely to the current, substrate, and nutrients (Table 24).

Discussion

Differences in generic richness and subfamily densities were associated with environmental characteristics in mesohabitats. Pools were the most chironomid diverse and dense habitats in this study, which is supported by previous work (Ferrington et al.

1995). Despite highest densities and richness values in pools, most chironomid genera were only collected from a single mesohabitat. Similarly, among 66 chironomid taxa collected from five mesohabitats in Kansas springs, 48 taxa only occurred in a single mesohabitat (Ferrington et al. 1995). Surface-fed tributaries had 18 unique chironomid

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taxa whereas only 8 taxa were unique to spring-fed tributaries in this study. Pools contained the most mesohabitat-specific taxa, and eight mesohabitat-specific genera were found in each tributary type pool. Unique genera of surface-fed tributary pools were

Djalmabatista, Paraphaenocladius, Parakiefferiellia, Peudochironomus, Saethira,

Stenochironomus, Strictochironomus, and Zavrelimyia. Unique genera of spring-fed tributary pools were Labrundinia, Micropsectra, Natarsia, Paralauterborniella,

Paratendipes, Procladius, Tribelos, and Xylotopus. Many of these genera are considered common and organically tolerant across freshwater habitats, suggesting that pools may host environmentally tolerant genera (Ferrington et al. 1995). Riffles contained the fewest mesohabitat-specific taxa. Surface-fed riffles included Monopelopia and

Sublettea. Monopelopia was previously documented to be widespread and Sublettea is known only from lotic waters (Ferrington et al. 2008), which supports collections from riffles in this study. In spring-fed riffles, only the widespread genus Dicrotendipes was mesohabitat specific, although it is known from lentic habitats in other studies

(Ferrington et al. 2008). Poor mobility and passive drift, in part, explains the lotic occurrence and minimal density of Dicrotendipes in this study.

Inclusion of chironomid genera in the SCI bioassessment was not informative and did not alter SCI scores or interpretation of water quality, which is likely due to the assessment structure. The SCI was designed to make quick and straightforward interpretations based on macroinvertebrate samples from riffle mesohabitats (Bowles et al. 2007). However, riffles are not always available or the habitat with the most macroinvertebrates. For instance, chironomids were most abundant and taxonomically rich from pool mesohabitats in this study. As such, use of Chironomidae in

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bioassessments should be more influential in larger streams and rivers with deeper water, lower oxygen, and few macroinvertebrate taxa. In such rivers and deep lakes

Chironomidae can thrive. For this reason, chironomid genera have been valuable in bioassessments when riffles are not available (Diggins and Stewart 1998, Mandaville

2002), especially in paleolimnology and limnology (Ashe et al. 1987). The difficulty with Chironomidae in bioassessments is the need for taxonomic resolution at the genus or species levels in order to properly interpret water quality and ecological implications

(Koperski 2009). For example, the most common genus in tributaries of the Current and

Jacks Fork rivers was Polypedilum, which is known for its global distribution (see Ashe et al. 1987). As such, Polypedilum at the genus level offers little ecological information.

In contrast, rarely occurring genera cannot send detectable signals to the SCI because the index relies upon high densities in order to impact the ranked scoring system (Bowles et al. 2007). Therefore, bioassessments at the genus level are likely to not be impacted by the inclusion of Chironomidae as is supported by this study. Previous research supported

Chironomidae alone in bioassessments as a suitable substitution for the entire macroinvertebrate community (Wilson and McGill 1977). This possibility was supported by preliminary patterns in multivariate ordinations of mesohabitats in this study. As such, these data reinforce the need for continued research of the entire macroinvertebrate community in the Current and Jacks Fork rivers in order to fully understand habitat dynamics.

The inclusion of genera of Chironomidae in the analyses did not change the interpretation of community relationships. Bioassessment interpretations based on metrics such as taxonomic richness, the biotic index, Shannon’s Diversity Index, or

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dominance were less precise than on multivariate analyses (Rabeni and Wang 2001).

When only genera of Chironomidae were assessed in impaired streams, these metrics did not lead to the same interpretations of water quality as did interpretations with the entire macroinvertebrate community (Rabeni and Wang 2001). Community relations were consistent across tributary types in this study despite inclusion or exclusion of genera of

Chironomidae, which suggests that either a gap exists in the metric designs or in our understanding of the ecological relationships rather than in the data. Current ecological knowledge associates most chironomids with organically enriched sites, yet taxa can be found in all freshwater habitats even where nutrients are low and oxygen is high

(Armitage et al. 1995). A range of environmental tolerances is found in the

Chironomidae that has successfully been used to assess differences between organically enriched sites (sewage effluent and agriculture) and sites without enrichment. Riffles contained lower diversity of chironomids than did pools; however, Orthocladinae C/O group and Rheocricotopus were strongly associated with the riffle mesohabitat. Rather than simply using bioindicators, it has been suggested that metrics reflecting mesohabitat/habitat similarity and temporal stability in structure may be better indicators of degradation (Koperski 2009). Although patterns are evident in this study, further work is necessary to elucidate differences among mesohabitats in the Current and Jacks Fork river tributaries.

Multivariate exploratory techniques have been increasingly successful in early detection of environmental change. Further, these techniques have made use of

Chironomidae to detect subtle changes in macroinvertebrate community structure

(Wymer and Cook 2003). Whereas current assessments focus on organic pollution

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events, Chironomidae might be better suited to determine smaller-scale disturbance events pertaining to watershed land cover or channel form. Multivariate ordination plots of the Current and Jacks Fork river tributaries revealed no changes in community structure when genera of Chironomidae were included. However, this was likely a result of the range of tributaries selected, which included two reference streams and did not incorporate streams with varying degrees of degradation (see Bowles et al. 2007). These multivariate techniques can be improved upon when a better ecological understanding is available, which would enable a more precise biotic index for the Chironomidae.

Preliminary work on bioindicators using chironomids is beginning (Carew et al. 2007).

For example, Chironomus, Dicrotendipes, and Psectrotanypus varius (Fabricius) are common and a reflection of streams with little riparian cover and heavy agricultural pollution (Adriaenssens et al. 2004, Kleine and Trivinho-Strixino 2005).

Chironomidae were diverse and abundant members in the tributaries of the

Current and Jacks Fork rivers, which indicates this family is important to consider in all ecological studies of the region. Including chironomid genera significantly increased the total taxonomic richness in this study. Although chironomid richness values are similar across most regions in the U.S., in Ozark reference streams of this study 48 genera were collected in comparison to the previous list of 25 (Rabeni and Wang 2001). Densities of

Chironomidae made up as much of 73% of the samples, indicating their presence is not only prevalent but reliable for use in sampling methods. Additionally, niche theory supports the use of genera and species in characterizing ecosystem health. Thus,

Chironomidae might be suited to indices of habitat or substrate stability because of relationships with nutrients, as seen here, and poor mobility. The role of chironomids in

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nutrient cycling research, especially in pools where densities are highest, is worthy of investigation. Pools are also the richest sources of chironomid taxa; as such, these macroinvertebrates fill a critical role as connections between primary and secondary trophic levels. Both taxonomic richness and density of Chironomidae suggest that this largely ignored family should be at the forefront of taxonomic, systematic, and ecological research.

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160

Table 18. Chironomidae subfamilies and genera collected from Brown vacuum samples in the Current and Jacks Fork river tributaries, Missouri in July and September 2013.

Surface-fed Tributary Spring-fed Tributary Taxa Riffle Pool Veg Total Riffle Pool Veg Total Tanypodinae 35 65 40 140 6 27 25 58 Ablabesmyia 42 11 53 1 2 3 Djalmabatista 4 4 Labrundinia 2 2 2 6 1 1 Larsia 1 1 3 3 Monopelopia 1 1 Natarsia 1 1 Nilotanypus 14 7 21 Paramerina 1 2 3 Pentaneura 2 2 Procladius 4 2 6 3 3 Telopelopia 1 1 T group 18 10 12 40 6 13 15 34 Zavrelimyia 2 2 6 2 8 Orthocladinae 62 6 11 79 73 11 25 109 Corynoneura 3 2 2 7 1 1 2 C/O group 33 2 35 37 1 6 44 Heterotrissocladius 3 1 4 Parakiefferiellia 1 1 Parametriocnemus 7 1 3 11 2 1 6 9 Paraphaenocladius 1 1 Rheocricotpus 7 2 9 12 2 1 15 Synorthocladius 3 1 4 4 1 5 Thienemanniella 3 1 4 6 1 7 Tvetenia 1 1 8 4 12 Xylotopus 1 1 Chironominae 172 80 46 298 29 72 42 143 Chironomus 2 1 3 Cladotanytarsus 46 18 3 67 Cryptochironomus 1 5 6 12 Cryptotendipes 1 1 Dicrotendipes 1 1 Glyptotendipes 1 1 Micropsectra 2 2 2 6 3 3 Microtendipes 5 3 8 Pagastiella 3 1 4 Paracladopelma 3 1 4 Paralauterborniella 2 2 Paratanytarsus 3 1 4 8 Paratendipes 5 1 6 Phaenopsectra 1 2 2 5 4 1 5 Polypedilum 80 9 14 103 3 30 23 56 Pseudochironomus 2 2 Rheotanytarsus 4 2 7 13 23 4 27 Saetheria 1 1 Stempellinella 2 4 2 8 1 1 Stenochironomus 1 1 Strictochironomus 8 8 Sublettea 1 1 Tanytarsus 21 15 5 41 1 14 7 22 Tribelos 4 1 5 2 2 161

Table 19. Three-way analyses of variance testing among metrics or indices for macroinvertebrates collected from the Current or Jacks Fork rivers, Missouri in July 2013. Factors are genera of Chironomidae inclusion, tributary type, and mesohabitat. BI = biotic index, SCI = Stream Condition Index, H’ = Shannon’s diversity index, TR = taxonomic richness, Chiro = genera of Chironomidae inclusion.

Variable Effect NDF DDF F p BI Chiro 1 48 0.00 0.958 Tributary 1 48 0.65 0.426 Mesohabitat 2 48 3.68 0.033 * Chiro x Tributary x Mesohabitat 7 48 0.13 0.996 H' Chiro 1 48 1.85 0.180 Tributary 1 48 18.04 <0.001 *** Mesohabitat 2 48 2.66 0.081 Chiro x Tributary x Mesohabitat 7 48 0.46 0.857 SCI Chiro 1 48 0.04 0.843 Tributary 1 48 4.82 0.033 * Mesohabitat 2 48 2.07 0.137 Chiro x Tributary x Mesohabitat 7 48 0.62 0.737 TR Chiro 1 48 15.56 <0.001 *** Tributary 1 48 16.41 <0.001 *** Mesohabitat 2 48 1.64 0.205 Chiro x Tributary x Mesohabitat 7 48 0.37 0.915 * p ≤ 0.05 ** p ≤0.01 *** p ≤ 0.001

162

Table 20. Two-way analysis of variance comparing taxonomic richness of Chironomidae using factors of tributaries and mesohabitats for macroinvertebrates collected from the Current or Jacks Fork rivers, Missouri in July 2013. TR = taxonomic richness.

Variable Effect NDF DDF F p Chironomidae TR Tributary 1 24 5.56 0.027 * Mesohabitat 2 24 0.88 0.428 Tributary x Mesohabitat 2 24 0.31 0.738 * p ≤ 0.05 ** p ≤0.01 *** p ≤ 0.001

163

Table 21. Multi-response permutation procedure tests associated with nometric multidimensional scaling of macroinvertebrate communities from mesohabitats with genera of Chironomidae excluded or included in the Current and Jacks Fork rivers, Missouri in July 2013. Surface = surface-fed tributaries, Spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation.

Variable Source T A p Excluding genera Tributary -4.59 0.11 0.002 ** of Chironomidae Mesohabitat -11.66 0.07 <0.001 *** Surface R v. Surface P -5.34 0.05 <0.001 *** Surface R v. Surface V -0.02 0.00 0.411 Surface R v. Spring R -6.49 0.05 <0.001 *** Surface R v. Spring P -9.34 0.08 <0.001 *** Surface R v. Spring V -7.78 0.06 <0.001 *** Surface P v. Surface V -1.21 0.01 0.116 Surface P v. Spring R -9.86 0.09 <0.001 *** Surface P v. Spring P -4.22 0.04 0.002 ** Surface P v. Spring V -6.20 0.05 <0.001 *** Surface V v. Spring R -6.09 0.04 <0.001 *** Surface V. Spring P -5.38 0.04 <0.001 *** Surface V v. Spring V -4.31 0.03 0.001 ** Spring R v. Spring P -5.63 0.05 <0.001 *** Spring R v. Spring V -2.98 0.02 0.011 * Spring P v. Spring V -0.73 0.01 0.207 Including genera Tributary -4.58 0.11 0.002 ** of Chironomidae Mesohabitat -19.55 0.07 <0.001 *** Surface R v. Surface P -10.36 0.05 <0.001 *** Surface R v. Surface V -1.87 0.01 0.046 * Surface R v. Spring R -10.93 0.06 <0.001 *** Surface R v. Spring P -12.70 0.08 <0.001 *** Surface R v. Spring V -10.83 0.06 <0.001 *** Surface P v. Surface V -2.64 0.01 0.010 * Surface P v. Spring R -13.79 0.09 <0.001 *** Surface P v. Spring P -9.33 0.05 <0.001 *** Surface P v. Spring V -9.66 0.05 <0.001 *** Surface V v. Spring R -10.00 0.06 <0.001 *** Surface V. Spring P -8.73 0.05 <0.001 *** Surface V v. Spring V -6.90 0.04 <0.001 *** Spring R v. Spring P -7.53 0.05 <0.001 *** Spring R v. Spring V -4.39 0.03 <0.001 *** Spring P v. Spring V -0.99 0.01 0.156 * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001 164

Table 22. Multi-response permutation procedure tests associated with nonmetric multidimensional scaling of communities of Chironomidae in mesohabitats from the Current and Jacks Fork rivers, Missouri in July 2013. Surface = surface-fed tributaries, Spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation.

Variable Source T A p Genera of Chironomidae Mesohabitat -11.33 0.06 <0.001 *** Surface R v. Surface P -5.38 0.04 <0.001 *** Surface R v. Spring R -3.33 0.03 0.008 ** Surface R v. Spring P -2.92 0.02 0.008 ** Surface P v. Surface V -3.36 0.03 0.005 ** Surface P v. Spring R -11.85 0.12 <0.001 *** Surface P v. Spring P -4.49 0.04 0.001 ** Surface P v. Spring V -4.46 0.04 0.001 ** Surface V v. Spring R -8.89 0.08 <0.001 *** Surface V v. Spring P -3.02 0.02 0.008 ** Surface V v. Spring V -2.55 0.02 0.015 * Spring R v. Spring P -9.50 0.10 <0.001 *** Spring R v. Spring V -6.09 0.06 <0.001 *** * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001

165

Table 23. Two-way analyses of variance comparing subfamily densities of Chironomidae between tributaries and among mesohabitats from the Current and Jacks Fork rivers, Missouri in July and September 2013.

Variable Effect NDF DDF F p Chironominae Tributary 1 24 2.85 0.104 Mesohabitat 2 24 1.62 0.220 Tributary x Mesohabitat 2 24 3.69 0.040 * Orthocladinae Tributary 1 24 1.46 0.238 Mesohabitat 2 24 11.54 <0.001 *** Tributary x Mesohabitat 2 24 0.08 0.925 Tanypodinae Tributary 1 24 8.69 0.007 ** Mesohabitat 2 24 4.29 0.026 * Tributary x Mesohabitat 2 24 0.83 0.450 * p ≤ 0.05 ** p ≤0.01 *** p ≤ 0.001

166

Table 24. Multiple linear regressions of environmental variables in relation to densities of the 10 most common genera of Chironomidae in July and September 2013 from the Current and Jacks Fork rivers, Missouri. C/O = Cricotopus/Orthocladius, T = Thienemannimyia. Dashes indicate no variables were statistically significant.

Regression Taxon Variable F Coefficient R2 p Ablabesmyia Depth 9.18 3.54 0.3 0.001 *** Temperature 2.97 0.004 ** Organics 2.99 0.004 ** Width 2.38 0.020 * Constant -4.42 Cladotanytarsus ------C/O group Depth 6.82 2.93 0.27 0.004 ** Organics -2.91 0.005 ** Constant 1.14 Nilotanypus Temperature 7.65 3.25 0.15 0.002 ** Organics -2.38 0.020 * Constant -2.11 Parametriocnemus Velocity 5.21 2.74 0.11 0.008 ** Macrophytes 2.09 0.040 * Constant 0.23 Polypedilum ------Rheocricotopus ------Rheotanytarsus ------Tanytarsus Periphyton 6.75 2.44 0.14 0.020 * Embeddedness 2.14 0.040 * Constant -0.98 T group Width 11.3 3.36 0.11 0.001 ** Constant -0.18 * p ≤ 0.05 ** p ≤0.01 *** p ≤ 0.001

167

45 a 40 35 b 30 25 20 15 10

5 Taxonomic Richness (TR) Richness Taxonomic 0

Mesohabitats

Figure 62. Three-way analysis of variance comparing taxonomic richness of the macroinvertebrate community comparing analyses when genera either excluded or included from analyses in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of Chironomidae, in Chiro = including genera of Chironomidae. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

168

45 a 40 b 35 30 25 20 15 10 5

Taxonomic Richness (TR) Richness Taxonomic 0

Mesohabitats

Figure 63. Three-way analysis of variance comparing taxonomic richness of the macroinvertebrate community between tributaries in July and September 2013 from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of Chironomidae, in Chiro = including genera of Chironomidae. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

169

16 a 14

12 b

10

8

6

4

Taxonomic Richness (TR) Richness Taxonomic 2

0 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats

Figure 64. Two-way analysis of variance comparing taxonomic richness of Chironomidae between tributaries from the Current and Jacks Fork rivers, Missouri in July and September 2013. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Columns represent means of five replicates ± standard errors.

170

a 3 b

2.5

2

1.5

1

0.5

0 Shannon’s Diversity Index (H’) Index Diversity Shannon’s

Mesohabitats

Figure 65. Three-way analysis of variance comparing Shannon’s Diversity Index values between tributaries in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of Chironomidae, in Chiro = including genera of Chironomidae. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

171

a 7 ab 6 b 5

4

3

2 Biotic Index (BI) Index Biotic 1

0

Mesohabitats

Figure 66. Three-way analysis of variance comparing biotic index values among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of Chironomidae, in Chiro = including genera of Chironomidae. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

172

18 a 16 b 14 12 10 8 6 4

2 Stream Condition Index (SCI) Index Condition Stream 0

Mesohabitats

Figure 67. Three-way analysis of variance comparing Stream Condition Index among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation, ex Chiro = excluding genera of Chironomidae, in Chiro = including genera of Chironomidae. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

173

NMS 2 NMS

Tributary Types Surface-fed Spring-fed + Centroid NMS 1

Figure 68. Nonmetric multidimensional scaling of surface-fed and spring-fed tributary macroinvertebrates excluding genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 4.13, randomization p = 0.040.

174

NMS 2 NMS

Tributary Types Surface-fed Spring-fed + Centroid NMS 1

Figure 69. Nonmetric multidimensional scaling of surface-fed and spring-fed tributary macroinvertebrates including genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 9.21, randomization p = 0.016.

175

Mesohabitats Surface NMS 2 NMS Riffles Veg Pools

Riffles Spring Veg Pools + Centroid NMS 1

Figure 70. Nonmetric multidimensional scaling of surface-fed and spring-fed mesohabitat macroinvertebrates excluding genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 16.46, randomization p = 0.004.

176

Mesohabitats Surface NMS 2 NMS Riffles Veg Pools

Riffles Spring Veg Pools + Centroid NMS 1

Figure 71. Nonmetric multidimensional scaling of surface-fed and spring-fed mesohabitat macroinvertebrates including genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 16.61, randomization p = 0.004.

177

Mesohabitats Surface

NMS 2 NMS Riffles Veg Pools

Riffles Spring Veg Pools + Centroid NMS 1

Figure 72. Nonmetric multidimensional scaling of surface-fed and spring-fed mesohabitat genera of Chironomidae in July 2013 from the Current and Jacks Fork rivers, Missouri. Stress = 14.7, randomization p = 0.004.

178

300 a

) 2 250

200

150 b b bc 100 b c

50 Density (individuals 0.05 m 0.05 (individuals Density

0 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats

Figure 73. Two-way analysis of variance comparing Chironominae densities among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences reflect a log transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

179

140 )

2 a 120

100

80

60 b

40

20 Density (individuals 0.05 m 0.05 (individuals Density

0 Surface R Surface P Surface V Spring R Spring P Spring V Mesohabitats

Figure 74. Two-way analysis of variance comparing Tanypodinae densities between tributaries in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences reflect a square root transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors.

180

140

a ) 2 120

100

80 b ab

60

40

20 Density (individuals 0.05 m 0.05 (individuals Density

0 Surface R Spring R Surface P Spring P Surface V Spring V Mesohabitats

Figure 75. Two-way analysis of variance comparing Tanypodinae densities among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences reflect a square root transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

181

180 a )

2 160

140

120

100

80 b 60

40 b

Density (individuals 0.05 m 0.05 (individuals Density 20

0 Surface R Spring R Surface P Spring P Surface V Spring V Mesohabitats

Figure 76. Two-way analysis of variance comparing Orthocladinae densities among mesohabitats in July and September 2013 Brown vacuum samples from the Current and Jacks Fork rivers, Missouri. Surface = surface-fed tributaries, spring = spring-fed tributaries, R = riffles, P = pools, V = marginal vegetation. Differences reflect a square root transformation. Pairwise comparisons with Bonferroni adjustments. Columns represent means of five replicates ± standard errors. Note the different arrangement of axis labels.

182

25

20

15

10 Percentage of Sample (%) Sample of Percentage 5

0

Figure 77. Most abundant Chironomidae collected in July and September 2013 from tributaries in the Current and Jacks Fork rivers, Missouri. T = Thienemannimyia, C/O = Cricotopus/Orthocladius.

183

Appendix A. Macroinvertebrates collected in slack-Surber samples during July 2011 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Platyhelminthes 0 0 0 0 0 1 0 0 0 1 0 1 7 0 1 0 3 0 14 Hydra 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 Oligochaeta 0 1 8 2 6 0 4 3 2 38 0 22 6 6 12 1 3 44 158 Hydracarina 8 14 25 6 35 29 3 9 24 67 58 77 35 43 21 0 12 9 475 Copepoda 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Sphaerium 0 0 0 0 0 0 0 2 0 0 1 0 0 0 1 0 3 6 13 Ferrissia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 9 10 Physa 0 0 0 0 0 0 0 0 0 3 0 0 1 0 0 1 0 1 6 Amnicola 26 1 1 233 64 11 90 2 115 416 53 69 51 0 0 0 5 17 1154 Elimia 16 1 0 2 3 2 7 0 0 3 0 0 923 13 2 0 60 50 1082 Helisoma 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 2 Orconectes virilis Hagen 0 0 1 0 0 1 3 0 3 0 0 1 0 1 0 0 0 4 14 Isopoda 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1

Asellus occidentalis Williams 0 0 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 184 Asellus 0 0 0 0 0 0 0 0 0 0 0 0 0 3 3 0 0 0 6

Gammaridae 0 0 0 188 1 0 0 0 0 52 0 0 1 0 0 0 0 0 242 Bactrurus 0 0 0 0 0 0 0 0 0 18 0 0 0 0 0 0 0 0 18 Gammarus 0 7 1 225 0 4 0 0 0 0 0 0 113 0 0 0 0 3 353 Allocragonyx 0 0 0 0 0 0 0 0 0 27 0 0 0 0 0 0 0 0 27 Entomobryidae 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 4 Podura aquatica Willem 0 0 0 0 0 3 0 0 0 1 0 0 0 0 0 0 0 0 4 Coenagrionidae 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Argia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Gomphidae 0 0 0 0 0 0 6 0 1 0 0 0 0 18 1 3 0 6 35 Hagenius brevistylus Selys 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 1 3 Gomphus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Stylogomphus 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0 2 Ephemeridae 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Ephemera 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Anthopotamus 0 0 0 0 0 0 0 0 0 0 0 0 0 5 5 0 5 6 21 Caenidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 0 3

Appendix A. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Caenis 4 15 3 0 15 7 6 1 2 0 16 77 0 21 15 4 4 28 218 Tricorythodes 50 142 153 0 101 93 16 15 78 14 99 263 0 32 14 0 21 43 1134 Ephemerella 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Serratella 3 1 4 2 1 0 0 0 1 0 0 3 0 1 0 0 7 2 25 Heptageniidae 0 0 0 0 11 8 115 3 8 21 9 34 0 69 31 1 13 39 362 Rhithrogena 0 0 0 0 0 0 18 52 5 0 20 16 0 0 0 0 8 0 119 Heptagenia 0 0 0 0 0 0 0 0 0 8 2 5 0 0 0 0 0 0 15 Leucrocuta 0 0 0 0 4 0 11 1 8 2 0 15 0 35 11 0 4 7 98 Nixe 0 1 0 0 8 2 0 0 0 1 0 10 0 0 0 0 0 0 22 Stenonema 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Stenonema femoratum 2 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 3 Maccaffertium 20 17 24 0 50 5 25 0 27 2 1 39 0 107 59 2 26 42 446 Maccaffertium bednaricki MacCafferty 1 0 0 0 0 0 42 0 3 0 0 2 0 0 0 0 1 0 49 Maccaffertium mediopunctatum McDunnough 1 0 1 0 0 0 9 0 4 2 0 3 0 8 1 1 1 3 34 Stenacron 0 0 0 0 1 2 1 0 0 0 0 0 0 11 26 0 0 7 48 Leptophlebiidae 0 0 0 0 1 0 0 0 0 1 0 0 0 0 0 3 0 0 5 Baetidae 0 1 1 100 5 1 6 1 0 4 17 11 2 5 0 0 9 1 164 185 Acentrella 1 2 6 0 1 3 2 4 3 0 11 4 0 0 0 1 0 0 38

Baetis 4 6 18 46 3 1 46 15 34 0 7 25 126 2 3 3 1 0 340 Procloeon 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Centroptilum 0 0 0 0 3 7 0 0 0 0 0 0 0 0 0 0 0 0 10 Plauditus 0 1 0 1 2 2 1 2 23 0 1 0 0 53 8 0 210 60 364 Isonychia 46 2 20 0 13 0 10 3 6 0 0 18 0 25 0 2 49 1 195 Perlidae 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 5 6 13 Leuctridae 2 3 6 0 19 11 12 0 4 0 0 8 0 0 0 0 0 1 66 Zealeuctra 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 0 5 Pteronarcys 0 0 0 0 1 0 1 0 3 0 0 1 0 1 0 0 0 1 8 Neoperla 0 0 0 0 0 0 0 0 1 0 1 1 0 2 1 0 3 0 9 Perlesta 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Perlesta shubuta Stark 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 Acroneuria 0 0 0 0 3 0 0 0 1 0 0 0 0 6 0 0 1 0 11

Appendix A. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Acroneuria mela 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Helicopsyche 0 0 0 0 2 0 0 0 0 0 2 0 0 29 1 0 19 22 75 Glossosoma 0 0 0 0 0 0 0 0 0 0 0 6 14 0 0 0 14 3 37 Glossosoma intermedium Klapalek 1 2 0 41 1 1 2 0 10 1 12 14 0 0 0 0 0 0 85 Protoptila 0 0 1 0 0 0 0 2 1 0 2 1 0 0 0 0 0 0 7 Gerridae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 2 Hydropsychidae 0 2 24 0 3 1 0 0 9 0 4 37 0 1 0 0 22 17 120 Hydropsyche 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 40 0 41 Ceratopsyche 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 2 Ceratopsyche morosa (Hagen) 2 2 6 0 0 0 0 1 1 0 0 0 0 0 0 1 28 0 41 Ceratopsyche slossonae (Banks) 1 1 84 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 87 Ceratopsyche piatrix (Ross) 0 10 16 4 3 2 0 2 1 0 4 1 0 0 0 0 0 0 43 Cheumatopsyche 115 28 184 1 21 0 0 8 20 0 12 58 0 153 59 0 45 60 764 Potomyia flava 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 1 0 3 Hydroptilidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Hydroptila 0 0 0 2 0 0 0 0 0 1 3 0 1 12 10 0 4 5 38 Orthotrichia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 6 0 6 Psychomyia flavida Hagen 1 7 11 0 0 0 0 3 1 0 2 1 0 1 0 0 7 0 34

186 Paduniella nearctica Flint 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1

Polycentropus 0 0 0 0 7 0 2 0 0 0 0 3 0 5 1 0 0 5 23

Chimarra 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 4 Lepidostomatidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Lepidostoma 0 0 1 2 2 5 4 1 8 4 11 14 181 4 10 0 3 4 254 Setodes 0 0 0 0 0 0 0 0 0 0 0 0 0 2 2 0 1 0 5 Brachycentridae 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Brachycentrus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 12 0 12 Micrasema 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Rhyacophila banksi 0 0 0 7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 7 Corydalidae 1 0 7 0 0 1 2 0 5 0 3 5 0 1 0 2 0 0 27 Corydalus 5 0 7 0 0 0 0 0 2 0 0 0 0 1 1 1 9 0 26 Nigronia 1 0 5 0 1 2 3 0 6 0 1 1 4 3 0 0 5 2 34

Appendix A. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Sialis 0 0 0 0 3 0 0 0 0 0 0 0 0 0 1 0 0 0 4 Heteroceridae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 2 Elmidae 0 0 1 0 0 0 0 7 0 4 18 3 7 2 3 0 0 0 45 Dineutus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Optioservus 79 48 87 6 51 61 44 97 64 15 194 305 165 16 11 0 14 2 1259 Stenelmis 1 5 7 1 0 1 75 2 12 0 11 13 0 69 57 1 25 59 339 Neoelmis 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Dubriaphia 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 1 0 3 Ordobrevia 0 0 0 0 0 0 0 0 0 0 0 0 0 5 0 0 0 0 5 Gonielmis 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 4 0 5 Ancyronyx 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Microcylleopus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 2 Rhizelmis 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Psephenidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Psephenus 72 0 2 0 0 0 27 0 25 0 2 0 0 0 1 5 1 15 150 Ectopria 0 0 0 0 0 0 0 0 0 0 0 0 0 5 1 0 1 19 26 Sperchopsis 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1

187 Diptera 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Empididae 1 3 2 0 0 0 0 0 1 0 3 0 0 0 0 0 3 1 14 Hemerodromia 0 5 6 1 1 1 1 3 4 2 10 1 2 11 34 0 21 10 113 Clinocera 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 Chelifera 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 1 0 3 Simulium 6 1 10 40 0 0 1 5 5 0 1 6 4 1 0 3 49 4 136 Ceratopogonidae 0 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 Bezzia 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 2 Chironomidae 16 180 134 63 119 136 11 34 46 615 161 134 130 93 37 14 214 78 2215 Antocha 0 2 1 0 0 0 0 0 0 0 1 0 2 4 4 0 2 0 16 Hexatoma 2 0 0 0 0 0 0 0 0 0 2 0 0 3 0 0 0 0 7 Tabanus 0 0 0 0 0 0 1 0 0 0 0 0 0 2 0 1 3 3 10 Tipulidae 0 1 0 0 0 0 1 0 0 0 5 1 0 0 1 0 0 0 9 Ochthera 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Atherix 0 1 3 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 6 Tanyderidae 0 0 2 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 4 Dixa 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1

Appendix B. Macroinvertebrates collected in slack-Surber samples during January 2012 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Platyhelminthes 16 0 22 0 11 1 1 63 7 0 0 5 12 0 0 0 0 3 141 Brachyobdellida 0 0 0 0 0 0 0 0 0 0 0 6 0 0 0 0 3 0 9 Oligochaeta 0 5 3 8 31 10 4 29 4 34 0 16 7 14 14 11 21 44 255 Hydracarina 5 22 22 22 37 58 24 19 10 80 61 14 28 9 10 1 4 3 429 Corbicula 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 3 Sphaerium 0 1 2 0 0 0 0 3 0 1 3 4 0 0 0 0 5 14 33 Ferrissia 0 0 0 0 1 0 0 0 0 0 0 0 0 2 1 1 0 10 15 Physa 0 0 1 1 2 3 0 1 0 1 0 0 0 0 0 0 0 0 9 Hydrobia 0 1 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 1 4 Amnicola 0 0 0 41 1 3 1 17 0 48 5 4 460 1 0 0 0 0 581 Elimia 109 9 0 4 17 129 1 15 4 89 43 33 13 0 1 5 79 47 598 Orconectes virilis Hagen 1 0 0 0 1 1 1 1 0 3 1 2 0 3 3 2 6 2 27 Hyallela azteca 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 0 0 1 3 Asellus 0 0 0 0 0 0 0 0 0 0 0 0 4 3 1 1 0 3 12

Gammaridae 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 3 5 188 Gammarus 0 1 1 97 1 2 0 0 0 35 0 0 355 0 0 0 0 1 493

Gammarus pseudolimnaeus Fabricius 0 1 0 458 0 6 0 0 0 446 0 0 0 0 0 0 0 0 911 Entomobryidae 1 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 3 Coenagrionidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 4 Argia 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 2 6 9 Calopteryx 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Gomphidae 9 0 0 0 1 0 2 1 0 0 0 2 0 1 0 0 0 16 32 Ophiogomphus 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 2 Baetisca 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Ephemera 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Anthopotamus 0 0 0 0 0 0 1 1 1 0 6 0 0 4 9 0 8 8 38 Caenis 0 0 0 0 0 0 0 0 0 0 0 0 0 29 8 6 1 28 72 Tricorythodes 3 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 0 5 Ephemerella 2 147 93 148 0 0 215 32 35 0 165 81 0 212 185 0 189 61 1565 Ephemerella excrucians Walsh 0 3 2 0 0 0 1 1 2 0 2 0 0 0 0 0 0 0 11 Ephemerella subvaria (Hendrickson) 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 0 0 2

Appendix B. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Eurylophella 8 9 1 0 24 1 18 7 9 0 62 52 1 363 214 3 38 247 1057 Serratella 1 0 4 0 5 0 1 2 0 2 1 1 0 0 0 0 0 0 17 Heptageniidae 0 0 0 2 15 0 83 0 1 4 1 6 0 4 5 0 0 17 138 Rhithrogena 0 0 0 0 1 0 19 16 5 0 6 8 0 0 0 0 76 4 135 Stenonema femoratum (Say) 0 0 0 0 0 0 0 1 0 1 0 0 0 5 14 2 0 0 23 Maccaffertium 23 0 59 0 380 1 25 48 3 0 50 72 0 48 29 11 24 183 956 Maccaffertium bednaricki MacCafferty 0 0 0 0 12 0 6 3 1 0 4 3 0 0 0 0 0 6 35 Maccaffertium mediopunctatum McDunnough 16 1 0 0 1 0 38 0 0 0 0 0 0 2 0 0 1 4 63 Stenacron 2 0 0 0 12 5 0 0 0 0 0 2 0 24 10 2 0 7 64 Leptophlebiidae 0 0 0 0 0 0 0 0 0 0 0 0 0 1 3 1 3 0 8 Leptophlebia 0 0 0 0 0 0 0 0 0 0 0 3 0 0 2 5 2 3 15 Paraleptophlebia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 1 6 7 17 Baetidae 0 0 0 0 0 0 1 1 0 2 0 0 0 0 0 0 0 0 4

Acentrella 2 1 1 0 0 0 6 0 2 0 0 0 0 0 0 1 0 0 13 189 Baetis 8 29 50 229 3 0 5 11 7 0 5 4 102 0 0 0 2 1 456

Centroptilum 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Isonychia 85 0 22 0 122 0 16 18 0 0 17 17 0 1 0 1 11 17 327 Plecoptera 0 0 0 0 0 0 4 0 0 0 0 0 0 2 2 0 1 1 10 Perlidae 0 0 0 0 5 0 0 0 0 0 0 2 0 0 0 0 0 5 12 Capniidae 0 0 0 0 0 0 4 0 0 0 0 0 0 0 0 1 0 1 6 Allocapnia 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 7 0 2 10 Leuctridae 0 0 0 6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 6 Prostoia 47 0 5 0 0 0 1 0 0 0 0 2 0 0 0 0 0 0 55 Pteronarcys 2 14 0 0 4 0 32 6 5 0 101 4 0 0 0 0 3 3 174 Taeniopterygidae 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 3 Taeniopteryx 0 0 0 0 0 0 8 0 22 0 0 0 0 0 0 0 0 0 30 Strophopteryx fasciata (Burmeister) 0 1 0 0 0 0 10 0 5 0 0 0 0 0 0 0 0 0 16 Chloroperlidae 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Haploperla brevis (Banks) 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1

Appendix B. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Neoperla 0 0 0 0 0 0 5 0 3 0 0 0 0 0 0 1 5 5 19 Attaneuria ruralis (Hagen) 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Helopicus nalatus (Frison) 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Perlinella 0 0 0 0 0 0 13 0 0 0 0 0 0 4 7 0 0 1 25 Perlinella ephyre (Newman) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 1 3 Perlesta 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Perlesta browni Stark 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 Agnetina 0 0 0 0 2 0 4 0 0 0 1 0 0 0 0 0 0 0 7 Agnetina capitata Pictet 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 2 Agnetina flavescens (Walsh) 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Paragnetina 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Perlodidae 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 Acroneuria internata (Walker) 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Isoperla dicala 0 2 3 0 5 0 0 7 1 0 9 6 0 0 0 0 0 0 33 Helicopsyche 0 0 0 0 0 0 0 0 0 4 8 0 0 0 0 0 2 0 14 Helicopsyche borealis (Hagen) 0 0 0 0 32 17 0 12 5 10 1 27 0 0 0 0 0 0 104 Glossosoma intermedium (Klapalek) 0 1 0 17 3 0 0 13 1 0 18 3 23 0 0 0 2 0 81

Agapetus 1 0 0 0 0 0 0 3 0 0 5 1 0 0 0 0 0 0 10 190 Protoptila 0 2 0 0 1 0 0 10 3 0 20 6 0 0 0 0 0 0 42

Neoplea striola (Fieber) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Hydropsyche scalaris Hagen 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Ceratopsyche 0 1 6 5 3 0 0 15 1 4 14 5 0 0 0 0 0 0 54 Ceratopsyche morosa (Hagen) 3 0 8 0 2 0 7 1 0 0 7 3 0 0 0 0 1 1 33 Ceratopsyche slossonae (Banks) 1 13 250 1 15 0 0 3 0 1 3 4 0 0 0 0 0 0 291 Ceratopsyche piatrix (Ross) 0 24 51 68 25 5 2 76 3 30 106 25 0 0 0 0 0 0 415 Cheumatopsyche 129 62 323 1 192 0 37 36 12 0 66 38 0 1 5 0 1 42 945 Hydroptila 0 1 0 1 0 3 35 1 1 0 1 0 0 2 0 0 4 4 53 Psycomyia flavida Hagen 0 82 68 0 25 39 10 40 13 16 85 43 0 4 7 0 27 25 484 Paduniella nearctica Flint 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 Polycentropus 2 0 0 0 0 2 0 0 0 0 1 0 0 3 15 0 0 11 34 Chimarra 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 0 2

Appendix B. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Lepidostoma 0 0 0 8 1 0 6 15 2 14 43 9 252 1 0 0 0 2 353 Nectopsyche 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Setodes oxapius (Ross) 0 0 0 0 0 0 0 0 5 0 0 0 0 0 0 0 0 0 5 Brachycentrus lateralis (Say) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 3 Pycnopsyche 0 0 0 0 1 1 0 1 0 0 0 0 0 0 0 1 0 0 4 Rhyacophila banksi Ross 0 0 0 5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 Neophylax 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 Corydalus 6 0 2 0 0 0 2 0 1 0 0 0 0 0 0 0 0 0 11 Nigronia 1 0 0 3 0 0 0 0 0 1 1 0 3 0 0 0 0 0 9 Sialis 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 2 Coleoptera 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Elmidae 26 162 203 1 141 1 4 166 40 11 323 71 0 0 2 1 0 6 1158 Optioservus 65 324 226 18 120 5 29 364 330 63 780 188 168 12 9 1 54 36 2792 Stenelmis 3 0 0 0 0 0 2 0 3 0 1 0 0 3 7 1 3 10 33 Dubriaphia 0 0 0 0 0 0 0 0 0 0 0 1 0 2 0 0 0 23 26 Ordobrevia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Microcylleopus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1

Psephenus herricki DeKay 37 0 0 0 0 0 8 0 12 0 1 2 0 2 1 9 4 22 98 191 Ectopria 0 0 0 0 0 0 0 0 0 0 1 0 0 2 4 0 5 19 31

Diptera 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Hemerodromia 3 8 4 0 3 0 7 1 12 0 1 1 0 0 0 0 3 5 48 Clinocera 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 Chelifera 0 4 1 0 0 0 7 0 0 0 0 0 0 0 0 0 0 0 12 Roederiodes 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Prosimulium 2 0 0 0 0 0 0 0 0 0 0 0 0 0 1 41 1 0 45 Simulium 34 2 15 2 3 0 1 0 0 0 0 0 0 0 0 2 0 0 59 Chironomidae 69 86 178 18 141 263 181 111 25 34 85 68 6 235 118 13 14 57 1702 Antocha 0 4 10 0 18 0 7 11 0 12 5 3 5 12 5 0 1 6 99 Hexatoma 0 0 0 0 0 0 27 0 4 0 0 0 0 0 0 0 2 0 33 Tabanus 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 0 2 0 4 Tipulidae 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1

Appendix B. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Tipula 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 2 Atherix 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Tanyderidae 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 Dixidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 2

192

Appendix C. Macroinvertebrates collected in upstream movement samplers during July 2011 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Hydracarina 0 0 0 0 0 0 0 2 1 0 0 0 0 0 0 0 0 0 3 Amnicola 1 0 0 11 0 2 2 0 1 1 0 6 6 0 0 0 0 0 30 Elimia 1 1 0 3 0 0 0 0 0 0 1 0 0 0 0 0 0 0 6 Gammarus 0 0 0 3 0 0 0 2 0 0 0 0 0 0 0 0 0 0 5 Gammarus pseudolimnaeus Fabricius 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 2 Allocragonyx 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 2 Entomobryidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Sminthuridae 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Caenis 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 3 Tricorythodes 0 1 0 0 3 0 1 3 2 0 4 1 0 1 0 0 0 0 16 Serratella 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Rhithrogena 0 0 0 1 1 0 1 3 0 0 5 2 0 0 0 0 0 0 13 Leucrocuta 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Maccaffertium 0 0 0 0 0 0 0 0 0 0 2 0 0 1 0 0 0 0 3

193 Maccaffertium bednaricki MacCafferty 0 0 0 0 0 0 1 0 2 0 0 0 0 0 0 0 0 0 3 Stenacron 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 0 0 0 3

Baetidae 0 0 0 2 2 0 0 0 0 0 2 0 1 1 0 0 2 1 11 Acentrella 0 0 1 0 1 0 0 0 0 0 0 3 0 0 0 0 1 0 6 Baetis 1 1 2 8 0 0 1 2 1 0 0 1 6 0 0 0 0 0 23 Plauditus 0 2 0 0 1 1 2 6 6 0 2 2 0 0 0 0 1 1 24 Isonychia 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 2 Perlesta decipiens (Walsh) 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Trichoptera 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 2 Glossosoma intermedium (Klapalek) 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 2 Rhagovelia 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Hebrus 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Ceratopsyche 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Ceratopsyche morosa (Hagen) 0 0 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 2

Appendix C. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Ceratopsyche slossonae (Banks) 0 0 4 0 1 0 0 1 0 0 1 0 0 0 0 0 0 0 7 Ceratopsyche piatrix (Ross) 2 4 5 1 5 1 0 4 0 0 5 0 0 0 0 0 0 0 27 Cheumatopsyche 0 0 0 0 1 0 0 0 4 0 2 4 0 0 0 0 0 0 11 Psychomyia flavida Hagen 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Lepidostoma 0 0 0 2 1 0 0 1 0 0 7 10 4 0 0 0 0 0 25 Nectopsyche 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 Triaenodes 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Corydalidae 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 Corydalus 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Nigronia 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Dineutus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 Optioservus 1 1 2 0 2 1 0 2 2 0 0 1 0 0 0 0 0 0 12 Stenelmis 0 2 1 0 1 0 0 0 0 0 1 2 0 1 0 0 0 0 8 Psephenus herricki (DeKay) 0 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0 0 0 2 Simulium 0 1 3 0 1 0 0 0 2 0 1 1 0 0 0 0 0 0 9 Ceratopogonidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 6 6 Atrichopogon 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 194 Chironomidae 0 5 3 4 9 2 0 8 8 3 40 1 5 1 0 0 4 4 97

Appendix D. Macroinvertebrates collected in upstream movement samplers during January 2012 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Hydracarina 0 0 0 0 1 24 1 1 3 3 0 0 0 0 0 0 0 0 33 Physa 0 0 0 0 2 1 0 0 0 0 0 0 0 0 0 0 0 0 3 Amnicola 0 0 0 4 0 0 0 0 0 0 0 0 1 0 0 0 0 0 5 Elimia 0 2 1 0 1 1 0 1 3 1 4 5 0 4 7 0 0 2 32 Orconectes virilis Hagen 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Gammarus pseudolimnaeus Fabricius 0 0 1 10 0 0 0 4 0 0 0 0 0 0 0 0 0 0 15 Anthopotamus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Ephemerella 0 0 1 0 0 0 0 3 0 0 1 2 0 1 1 0 1 0 10 Ephemerella excrucians Walsh 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Eurylophella 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 2 Maccaffertium mediopunctatum McDunnough 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Baetis 0 0 0 6 0 0 0 3 0 0 0 0 1 0 0 0 0 0 10 Isonychia 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Prostoia 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Taeniopterygidae 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1

Strophopteryx fasciata (Burmeister) 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 195 Perlinella drymo (Newman) 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1

Helicopsyche borealis (Hagen) 0 2 0 0 1 0 0 1 0 0 0 10 0 0 0 0 0 0 14 Glossosoma intermedium (Klapalek) 0 0 0 2 0 0 0 0 0 0 0 2 0 0 0 0 0 0 4 Protoptila 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 2 Ceratopsyche morosa (Hagen) 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 2 0 3 Ceratopsyche slossonae (Banks) 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Ceratopsyche piatrix (Ross) 0 0 0 11 0 0 0 8 0 1 0 1 1 1 0 0 0 0 23 Hydroptila 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Psychomyia flavida Hagen 0 0 0 0 0 0 0 3 0 0 0 6 0 0 0 0 0 0 9 Lepidostoma 0 0 0 0 0 8 0 0 2 0 0 1 4 0 0 0 0 0 15 Pycnopsyche 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Neophylax 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2

Appendix D. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Corisella 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Lutrochus luteus LeConte 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Optioservus 0 1 2 0 0 0 1 3 1 0 0 2 2 0 0 0 0 0 12 Ectopria 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Chironomidae 5 2 0 0 0 0 2 0 0 0 0 7 0 3 0 1 0 0 20 Antocha 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1

Dixidae 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1

196

Appendix E. Macroinvertebrates collected in drift samplers during July 2011 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Nematomorpha 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 Oligochaeta 0 1 0 1 0 0 0 0 0 2 0 0 2 0 0 3 0 0 9 Hydracarina 0 3 1 2 4 12 2 13 5 22 11 5 8 3 2 1 2 4 100 Ferrissia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 0 3 Physa 1 0 0 1 0 2 0 0 0 0 0 0 1 0 0 2 0 1 8 Hydrobia 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 2 Amnicola 3 0 0 117 1 2 15 0 5 28 1 3 236 0 0 0 0 0 411 Elimia 7 0 0 1 0 0 12 2 4 2 2 6 61 0 1 2 1 10 111 Helisoma 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Orconectes virilis Hagen 1 1 1 1 0 2 0 0 1 0 0 1 0 0 0 0 0 0 8 Asellus occidentalis Williams 0 2 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 Hyallela azteca (Saussure) 3 0 0 0 0 3 0 1 1 2 0 1 0 2 2 2 1 1 19 Gammaridae 0 0 0 6 0 0 0 0 0 0 0 0 12 0 0 0 0 0 18 Gammarus 1 4 0 99 0 1 0 1 0 0 1 0 0 0 0 0 0 0 107 Gammarus pseudolimnaeus Fabricius 0 0 0 0 0 0 0 0 0 54 0 0 0 0 0 0 0 0 54

197 Allocragonyx 0 0 0 0 0 0 0 0 0 0 0 0 87 0 0 0 0 0 87 Coenagrionidae 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1

Argia 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 2 Calopterygidae 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 2 Hetaerina 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 0 2 Macromiidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 2 Gomphidae 2 1 0 0 0 0 2 0 2 0 0 0 0 11 7 2 36 41 104 Hagenius brevistylus Selys 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 2 0 0 3 Ophiogomphus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Gomphus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 2 Stylogomphus 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 Anthopotamus 0 0 0 0 0 0 1 0 1 0 1 0 0 0 0 1 0 0 4 Caenidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 19 8 29 Caenis 12 6 8 0 3 2 2 2 2 0 5 5 0 0 0 0 0 0 47 Tricorythodes 62 75 46 0 6 11 28 30 48 2 16 21 0 2 2 17 4 5 375 Ephemerella 0 0 0 0 0 0 0 0 1 0 0 1 0 5 2 1 28 8 46

Appendix E. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Eurylophella 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Eurylophella aestiva (McDunnough) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Serratella 9 3 1 1 3 4 3 3 0 0 2 3 0 0 0 0 1 1 34 Heptageniidae 0 1 0 0 0 0 79 8 14 1 7 9 0 0 0 0 1 0 120 Rhithrogena 0 3 0 0 79 26 29 126 58 0 51 70 0 2 2 0 8 9 463 Heptagenia 0 0 0 0 0 0 0 0 0 39 0 0 0 0 0 0 0 0 39 Leucrocuta 1 0 0 0 0 0 29 3 19 0 19 19 0 1 15 1 21 28 156 Nixe 0 0 0 0 0 2 0 0 0 0 8 0 0 5 0 0 1 5 21 Stenonema 0 0 0 0 0 0 0 0 0 0 0 0 0 2 1 0 1 3 7 Stenonema femoratum (Say) 45 0 3 0 2 0 3 2 2 0 0 0 0 0 0 1 0 0 58 Maccaffertium 8 6 1 0 4 2 36 2 18 0 4 12 0 0 0 7 0 1 101 Maccaffertium bednaricki MacCafferty 0 0 0 0 0 0 19 0 4 0 0 0 0 3 5 6 13 14 64 Maccaffertium mediopunctatum McDunnough 2 0 0 0 0 0 26 0 2 0 1 0 0 0 0 0 0 2 33 Stenacron 0 2 0 0 1 1 340 4 84 0 13 9 0 0 1 2 0 2 459 Leptophlebiidae 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 Choroterpes 7 0 0 0 2 0 0 2 0 1 1 2 0 0 0 8 0 0 23 Baetidae 0 2 1 25 0 0 0 27 32 5 11 8 0 1 0 1 2 0 115

Paracloeodes 0 0 0 0 0 0 0 0 0 0 0 0 10 128 5 6 183 11 343 198 Acentrella 2 16 7 0 10 5 7 17 14 0 12 12 0 0 0 0 0 1 103

Baetis 9 44 14 787 32 8 22 38 29 23 20 56 0 1 0 4 1 20 1108 Baetis flavistriga McDunnough 0 0 0 0 0 0 0 0 0 0 0 0 204 0 0 6 7 3 220 Procloeon 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 1 0 0 3 Centroptilum 0 0 0 1 1 0 0 0 0 6 0 0 0 0 0 0 0 0 8 Plauditus 1 26 7 0 42 45 9 55 59 0 46 32 0 67 26 10 330 379 1134 Isonychia 192 0 0 0 10 5 3 1 8 0 11 15 0 0 0 0 0 0 245 Pteronarcys 0 0 0 0 1 1 0 0 1 0 1 2 0 0 0 0 0 0 6 Isoperla mohri Frison 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 2 Neoperla 0 0 0 0 0 0 6 0 8 0 4 2 0 0 0 0 0 0 20 Neoperla osage Stark and Lentz 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 Perlinella ephyre (Newman) 0 0 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 Perlesta 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 3 3 9

Appendix E. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Perlesta shubuta Stark 1 0 0 0 0 1 0 5 4 0 0 2 0 0 0 0 0 0 13 Perlesta decipiens (Walsh) 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Acroneuria 0 0 0 0 0 2 1 0 0 0 0 0 0 0 0 0 2 0 5 Petrophila 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Helicopsyche 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 4 0 0 5 Glossosoma 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 2 Glossosoma intermedium (Klapalek) 0 0 0 4 0 0 0 1 0 0 0 1 0 0 0 0 0 0 6 Neoplea 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Gerridae 0 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 3 Rhagovelia 1 0 2 0 0 0 7 0 0 0 0 0 0 0 0 0 0 0 10 Microvelia 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Hydropsychidae 0 5 1 0 4 2 0 3 0 0 2 2 0 0 0 0 0 0 19 Hydropsyche 0 0 0 0 0 1 0 1 0 0 0 0 0 0 1 5 6 5 19 Ceratopsyche 0 0 0 0 0 0 2 4 2 0 1 0 0 0 0 0 1 2 12 Ceratopsyche morosa (Hagen) 5 3 0 0 5 1 1 6 8 0 6 7 0 1 2 1 2 1 49 Ceratopsyche slossonae (Banks) 11 76 39 0 21 19 0 16 7 0 15 10 0 0 0 0 0 0 214 Ceratopsyche piatrix (Ross) 0 47 9 0 37 26 0 62 29 4 20 19 0 0 0 0 0 0 253

199 Cheumatopsyche 69 15 8 0 2 5 2 24 18 0 28 17 0 0 0 0 6 1 195 Potomyia flava (Hagen) 0 0 0 0 0 0 0 0 0 0 2 0 0 2 3 4 19 27 57

Hydroptila 0 0 0 11 0 0 1 5 0 0 1 0 0 1 0 0 1 0 20 Orthotrichia 0 0 0 0 0 0 0 0 0 0 0 0 1 2 1 2 1 1 8 Ithytrichia clavata Morton 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Oxyethira 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Psychomyia flavida Hagen 1 0 2 0 3 0 0 3 3 0 2 1 0 2 0 0 0 0 17 Lype diversa (Banks) 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Polycentropus 2 2 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 6 Chimarra 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Lepidostomatidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 0 0 5 Lepidostoma 0 3 3 12 12 6 7 98 30 0 46 65 0 0 0 0 0 0 282 Leptoceridae 0 0 0 0 0 0 1 0 0 0 0 0 310 3 7 0 4 9 334 Nectopsyche 0 0 0 0 0 1 1 7 2 0 1 0 0 0 0 1 0 1 14

Appendix E. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Setodes 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 5 1 3 13 Oecetis 8 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 8 Triaenodes 0 0 0 0 0 1 0 0 1 0 0 0 0 0 0 3 2 1 8 Brachycentrus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 2 Brachycentrus lateralis (Say) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Micrasema 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Rhyacophila banksi Ross 0 0 0 8 0 0 0 0 0 0 0 0 0 0 0 0 0 0 8 Corydalidae 5 0 0 0 0 0 7 1 2 0 7 0 0 0 0 0 0 0 22 Corydalus 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 9 0 0 12 Nigronia 0 0 1 0 1 1 0 0 0 0 0 1 0 0 0 7 0 0 11 Sialis 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 2 Heteroceridae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Corisella 0 0 0 0 12 4 0 0 1 0 2 1 0 0 0 0 0 0 20 Enochrus 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 2 Berosus 0 2 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 4 Oreodytes 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 2 Laccodytes 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 2

Laccornis 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 2 200 Neoporus 0 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 2

Elmidae 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 2 Dineutus 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 1 1 4 Gyrinus 1 0 8 0 0 0 0 0 0 0 0 0 0 3 0 0 1 2 15 Oulimnius 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Macronychus 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Cleptelmis 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Optioservus 16 42 35 10 23 39 11 122 62 9 16 18 0 0 0 0 1 0 404 Stenelmis 8 10 5 0 3 3 52 32 48 0 8 16 16 1 0 27 6 8 243 Dubriaphia 0 0 0 0 0 2 0 0 2 0 0 1 0 4 3 11 16 11 50 Ordobrevia 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 1 4 3 10 Psephenidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 2 Psephenus herricki (DeKay) 10 0 1 0 0 0 1 0 0 0 1 0 0 0 0 0 0 0 13

Appendix E. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Ectopria 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 17 0 0 18 Sperchopsis 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 7 0 1 8 Empididae 1 1 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 Hemerodromia 0 1 0 0 0 0 1 1 0 0 3 0 0 0 0 0 1 1 8 Clinocera 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 2 5 Chelifera 0 0 0 1 0 0 0 1 0 1 0 0 1 0 0 0 0 0 4 Simuliidae 0 0 0 0 0 0 0 0 0 0 0 0 5 0 0 0 0 0 5 Simulium 3 34 38 17 17 3 10 74 15 1 12 15 0 0 0 2 0 0 241 Ceratopogonidae 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 14 3 2 21 Chironomidae 14 171 126 109 109 80 17 262 113 347 204 56 0 0 0 0 1 0 1609 Antocha 0 2 1 0 0 0 0 0 0 0 0 0 399 93 43 172 183 81 974 Hexatoma 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 3 Tabanus 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Tipulidae 0 2 0 0 2 0 0 1 0 0 0 0 0 0 0 0 0 0 5 Atherix 0 1 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 3

201 Dixidae 1 4 1 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 8 Dixa 0 0 0 0 0 0 0 0 0 0 0 0 0 7 3 0 0 0 10

Appendix F. Macroinvertebrates collected in drift samplers during January 2012 from the Current River, Missouri. T = tributary, UCR = upstream of tributary in the Current River, DCR = downstream of tributary in the Current River. Values are sums of original data from three subsamples.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Platyhelminthes 1 0 0 1 0 0 0 1 1 0 0 1 0 0 0 0 0 0 5 Nematomorpha 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 1 0 4 Oligochaeta 0 0 0 0 0 0 2 3 1 0 0 0 1 0 0 2 0 0 9 Hydracarina 2 2 3 4 26 6 2 3 0 7 4 0 22 0 0 0 0 2 83 Sphaerium 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Ferrissia 2 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 4 Physa 0 0 0 7 1 0 0 1 0 1 0 0 3 0 0 0 0 0 13 Hydrobia 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Amnicola 0 0 0 90 0 0 1 3 0 0 1 0 208 0 0 0 0 0 303 Elimia 2 0 1 0 3 16 4 17 8 10 11 48 10 2 4 0 12 6 154 Helisoma 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 2 Orconectes virilis Hagen 1 0 0 0 0 1 0 0 0 0 0 0 0 0 0 3 0 0 5 Hyallela azteca (Saussure) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Asellus 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 Gammaridae 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 202 Gammarus 0 1 0 1 0 0 0 0 0 2 0 0 163 0 0 0 0 0 167

Gammarus pseudolimnaeus Fabricius 0 1 2 66 0 0 0 1 1 34 2 0 79 0 0 0 0 0 186

Entomobryidae 5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 Poduridae 5 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 6 Coenagrionidae 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 2 Argia 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Calopteryx 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 Hetaerina 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 2 Gomphidae 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 2 Hagenius 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 2 Stylogomphus 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Baetisca 1 0 0 0 0 0 0 0 0 0 0 0 0 4 0 0 1 0 6 Ephemera 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Anthopotamus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Caenis 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 2 Tricorythodes 1 11 3 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 16

Appendix F. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Ephemerella 0 117 50 161 6 0 106 30 28 1 148 86 0 405 321 0 46 10 1515 Ephemerella excrucians Walsh 0 4 3 29 0 0 0 8 4 3 3 2 0 0 0 0 0 0 56 Ephemerella subvaria (Hendrickson) 0 1 1 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 4 Eurylophella 1 1 2 0 3 0 15 1 2 0 7 4 0 36 0 0 9 10 91 Eurylophella aestiva (McDunnough) 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Heptageniidae 0 0 0 1 0 0 0 1 3 0 0 0 0 1 0 0 0 0 6 Rhithrogena 0 0 0 0 0 0 2 32 4 0 7 20 0 6 0 0 2 1 74 Stenonema femoratum (Say) 4 1 0 0 0 0 1 0 1 0 0 0 0 0 0 2 0 0 9 Maccaffertium 10 2 20 0 9 0 12 6 3 1 6 10 0 1 0 1 2 7 90 Maccaffertium bednaricki MacCafferty 0 0 0 0 0 0 6 5 7 0 0 1 0 0 0 0 0 0 19 Maccaffertium mediopunctatum McDunnough 18 0 0 0 0 0 19 0 12 0 0 0 0 0 0 0 0 0 49 Stenacron 0 0 1 0 6 0 1 11 8 0 8 19 0 3 0 0 4 2 63 Leptophlebia 2 7 33 0 3 0 29 9 76 0 9 3 0 3 0 4 42 13 233 Paraleptophlebia 0 0 0 0 0 0 0 0 1 0 1 1 0 5 0 0 11 3 22 Acentrella 3 0 1 0 1 0 4 2 0 0 1 3 0 0 0 1 0 0 16 Baetis 9 146 153 224 26 0 1 30 13 12 12 28 61 0 0 0 3 0 718 Plauditus 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 2

203 Isonychia 7 5 17 0 4 0 13 12 4 0 18 19 0 9 3 1 5 7 124

Plecoptera 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 2

Capniidae 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 2 0 0 3 Allocapnia 6 0 1 0 0 0 29 0 17 0 0 2 0 1 0 4 0 0 60 Allocapnia granulata (Claassen) 13 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 13 Nemouridae 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Shipsa rotunda (Claassen) 0 0 0 0 0 0 0 0 0 0 1 0 0 3 0 0 0 0 4 Prostoia 47 2 4 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 54 Prostoia completa (Walker) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 Pteronarcys 0 3 1 0 1 0 6 2 1 0 4 4 0 0 0 0 0 0 22 Taeniopteryx 0 0 0 0 0 0 6 1 4 0 2 0 0 0 0 0 0 0 13 Taeniopteryx lita Frison 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Taeniopteryx metequi Ricker and Ross 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Strophopteryx fasciata (Burmeister) 0 1 0 0 0 0 21 0 4 0 1 3 0 0 0 0 0 0 30

Appendix F. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Haploperla brevis (Banks) 0 0 0 0 0 0 0 0 1 0 1 1 0 0 0 0 0 0 3 Neoperla 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 0 2 Perlinella drymo (Newman) 1 0 1 0 0 0 11 4 23 0 0 0 0 0 0 0 0 0 40 Perlinella ephyre (Newman) 0 1 0 0 0 0 3 0 2 0 0 0 0 0 0 0 0 1 7 Agnetina 0 0 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 3 Agnetina flavescens (Walsh) 0 0 0 0 0 0 1 0 2 0 0 0 0 0 0 0 0 0 3 Paragnetina 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Acroneuria internata (Walker) 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Isoperla dicala Frison 0 3 1 0 2 0 1 12 2 0 4 2 0 0 0 0 1 0 28 Lepidoptera 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 Petrophila 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Helicopsyche 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 3 Helicopsyche borealis (Hagen) 0 0 0 0 1 0 0 11 1 0 3 2 0 0 0 0 0 0 18 Glossosoma intermedium (Klapalek) 0 0 0 1 0 0 0 10 2 0 1 0 15 0 0 0 0 0 29 Agapetus 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Protoptila 0 0 0 0 0 0 0 10 1 0 0 1 0 0 0 0 0 0 12 Hydropsychidae 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1

204 Hydropsyche scalaris Hagen 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 3 Ceratopsyche 0 0 0 0 0 0 0 2 0 0 0 5 0 0 0 0 0 0 7

Ceratopsyche morosa (Hagen) 1 0 2 0 0 0 1 4 2 0 1 3 0 0 0 0 4 0 18 Ceratopsyche slossonae (Banks) 1 30 32 0 3 0 0 9 0 0 2 4 0 0 0 0 0 0 81 Ceratopsyche piatrix (Ross) 0 58 23 63 11 1 0 164 9 14 23 25 0 0 0 0 0 0 391 Cheumatopsyche 18 16 33 0 3 0 6 16 2 0 3 5 0 0 0 0 1 1 104 Hydroptila 0 0 0 0 0 0 32 0 7 0 0 0 0 0 0 0 9 1 49 Psychomyia flavida Hagen 1 27 37 0 8 0 0 19 4 0 13 29 0 1 0 0 2 11 152 Polycentropus 0 0 0 0 1 0 0 0 0 0 0 2 0 0 0 0 0 0 3 Chimarra 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Lepidostoma 0 0 0 13 0 0 2 18 5 4 2 12 207 0 0 0 0 1 264 Nectopsyche 0 0 0 0 1 0 0 1 0 0 0 0 0 0 0 0 0 0 2 Setodes oxapius (Ross) 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Oecetis 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1

Appendix F. Continued.

Big Creek Pulltite Spring Sinking Creek Round Spring Blue Spring Rocky Creek Taxa T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR T UCR DCR Total Brachycentrus 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Brachycentrus lateralis (Say) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 1 3 Pycnopsyche 3 0 15 0 2 0 0 4 0 0 0 1 0 0 0 1 1 0 27 Rhyacophila banksi Ross 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Neophylax 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 Nigronia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Sialis 0 0 0 0 0 0 0 0 1 0 1 1 0 0 0 0 0 0 3 Hydrophilus 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Hydroporinae 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 3 Elmidae 3 0 2 0 1 0 0 10 0 0 6 8 0 0 0 0 0 0 30 Gyrinus 1 7 2 0 1 0 0 0 0 0 2 0 0 1 0 0 0 0 14 Optioservus 10 59 32 3 11 0 11 167 33 3 56 104 55 0 0 0 0 0 544 Stenelmis 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 2 Dubriaphia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Psephenus herricki (DeKay) 0 0 0 0 0 0 2 0 1 0 1 0 0 0 0 0 0 3 7 Ectopria 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Hemerodromia 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 Clinocera 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 2 205 Prosimulium 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 0 0 5

Simulium 13 6 10 0 0 0 0 0 1 3 0 0 0 0 0 0 1 0 34

Chironomidae 126 72 106 21 39 0 102 40 25 7 40 19 8 35 5 23 32 64 764 Antocha 0 7 4 0 2 0 1 0 0 0 1 0 1 0 0 0 2 0 18 Hexatoma 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 2 Tipula 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 Pilaria 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 Atherix 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 2 Dixidae 0 0 0 0 0 0 1 3 0 0 0 0 0 0 0 0 0 0 4

Appendix G. Macroinvertebrates collected July 2013 from surface-fed tributary mesohabitats in the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation. Values are sums of original data from three subsamples.

Ashley Creek Big Creek Sinking Creek Bay Creek Rocky Creek Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Platyhelminthes 4 0 0 4 0 1 0 0 0 2 1 2 0 0 2 16 Oligochaeta 1 96 47 1 34 9.5 1 12 6 25 14 15 9 9 51 331 Ferrissia 0 0 0 0 0 0 0 0 0 1 1 3 0 0 0 5 Heliosoma 0 0 0 1 1 0 0 0 0 0 0 0 0 0 1 3 Physa 2 0 0 0 1 0 0 0 0 0 0 2 0 0 0 5 Elimia 7 15 0 6 6 30 12 0 0 9 1 6 1 2 0 95 Amnicola 31 78 50 3 11 4 6 0 0 25 3 10 2 0 0 223 Fossaria 0 1 0 0 0 0 0 0 0 0 1 1 0 0 0 3 Sphaerium 0 1 0 0 1 1 0 0 0 0 0 0 0 7 0 10 Hydracarina 30 12 9 75 7 1 66 5 50 19 1 16 33 12 3 339 Copepoda 0 2 3 0 2 0 0 0 0 0 2 0 3 3 2 17 Ostracoda 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 2 Isopoda 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Asellus 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 2 206 Lirceus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Orconectes 0 1 2 1 1 3 0 0 0 1 1 1 1 0 0 12 Amphipoda 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Gammaridae 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 2 Stygobromus 0 0 0 1 0 0 0 0 0 0 0 7 0 0 0 8 Gammarus pseudolimnaeus Fabricius 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0 2 Hyallela azteca (Saussure) 0 2 0 1 0 7 0 2 0 0 2 0 2 12 75 103 Collembola 0 1 1 3 0 0 0 0 0 0 0 3 0 0 0 8 Podura aquatica Willem 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 Entomobryidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 2 Aniosoptera 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 2 Macromiidae 0 0 0 0 1 0 0 1 0 0 1 0 1 0 0 4 Macromia 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 2 Gomphidae 0 8 1 0 2 1 1 2 10 2 2 5 0 1 0 35

Appendix G. Continued.

Ashley Creek Big Creek Sinking Creek Bay Creek Rocky Creek Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Hagenius brevistylus Selys 0 2 0 0 0 0 0 1 0 0 0 0 0 0 0 3 Stylogomphus 1 2 0 0 0 1 1 5 1 5 5 4 0 0 0 25 Ophiogomphus 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Epiaeshna 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 Remartina 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Somatochlora 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 2 Zygoptera 0 1 1 0 0 0 0 0 1 0 0 0 0 0 0 3 Coenagrionidae 0 0 3 0 1 0 0 0 0 0 0 0 3 0 23 30 Calopterygidae 0 0 0 0 0 0 0 0 0 1 0 4 0 0 0 5 Hetaerina 0 0 0 0 0 1 0 2 0 0 0 0 0 0 0 3 Argia 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Plecoptera 1 0 0 0 0 1 1 0 1 0 0 0 0 0 0 4 Capniidae 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Allocapnia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

Leuctra 31 1 2 0 0 0 1 45 5 59 105 121 1 0 0 371 207 Perlidae 0 0 0 0 0 0 4 0 1 1 0 0 3 0 0 9 Neoperla 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Neoperla falayah Stark and Lentz 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Acroneuria evoluta Klapalek 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 2 Acroneuria internata (Walker) 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Perlinella 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Perlodidae 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 Baetisca 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Ephemeridae 3 4 0 0 1 0 0 0 0 0 0 0 0 0 0 8 Ephemera 0 3 0 0 0 0 0 4 0 0 0 0 0 0 0 7 Hexagenia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Rhithrogena 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0 2 Isonychia 84 0 4 13 0 0 1 0 10 20 0 28 3 0 0 163

Appendix G. Continued.

Ashley Creek Big Creek Sinking Creek Bay Creek Rocky Creek Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Isonychia bicolor (Walker) 0 0 0 0 0 0 0 0 6 0 0 0 0 0 0 6 Caenis 24 40 16 23 32 21 1 50 10 7 20 5 18 85 11 363 Tricorythodes 143 32 40 0 0 0 7 15 2 0 0 0 1 0 0 240 Ephemerella 5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 Heptageniidae 4 0 0 3 0 0 5 0 1 0 0 0 0 1 0 14 Stenacron 0 1 0 0 0 0 0 2 0 0 2 2 3 0 4 14 Maccaffertium 40 0 10 11 3 2 11 3 35 108 18 61 8 1 7 318 Macaffertium mediopunctatum McDunnough 8 0 0 0 0 0 2 0 5 0 0 0 2 0 0 17 Stenonema femoratum (Say) 2 1 1 0 0 1 0 0 0 0 2 0 1 1 17 26 Stenonema pulchellum (Walsh) 0 0 0 0 0 0 12 0 28 0 0 0 0 0 0 40 Leucrocuta 0 0 0 0 0 0 13 0 12 0 0 0 0 0 0 25 Leptophlebiidae 0 0 0 0 1 1 0 2 0 0 6 2 27 36 0 75 Leptophlebia 0 0 0 1 1 0 0 2 0 0 0 0 8 41 2 55 Paraleptophlebia 0 0 0 0 0 0 0 0 0 0 7 2 4 19 2 34 Serratella 7 0 0 0 0 0 1 0 5 0 0 0 0 0 0 13 208 Baetidae 50 0 13 8 1 4 1 0 18 2 0 0 3 3 4 107

Baetis 20 0 4 33 0 1 4 0 27 45 0 2 6 0 0 142 Centroptilum 0 0 0 0 0 1 0 0 0 0 1 0 0 0 2 4 Procloeon 0 2 0 0 0 0 0 13 0 0 0 0 0 5 0 20 Acentrella 15 1 1 12 0 5 1 0 1 2 0 1 6 0 0 45 Plauditus 0 0 0 0 0 0 0 0 2 1 0 0 0 0 0 3 Rhagovelia 0 0 1 0 0 1 1 0 0 0 0 1 0 0 0 4 Mesovelia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 3 Ranatra 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Saldidae 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Corydalidae 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 Sialis 0 0 0 0 0 0 0 0 0 0 1 0 0 0 3 4 Corydalus 0 0 0 0 0 0 0 0 0 0 0 0 4 0 0 4

Appendix G. Continued.

Ashley Creek Big Creek Sinking Creek Bay Creek Rocky Creek Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Nigronia 2 0 0 0 0 0 0 0 2 8 1 8 1 0 0 22 Trichoptera 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Helicopsyche 1 1 1 1 0 0 0 0 0 1 0 1 0 0 0 6 Helicopsyche borealis (Hagen) 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 Pycnospsyche 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Psychomyia flavida Hagen 3 0 0 2 0 0 0 0 0 0 0 0 0 0 0 5 Wormaldia 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Chimarra 30 0 0 14 0 0 1 0 9 3 0 1 15 0 0 73 Hydropsychidae 10 0 1 54 0 2 1 0 0 0 0 0 0 0 0 68 Cheumatopsyche 27 0 0 66 0 2 1 0 4 113 1 19 2 0 0 235 Ceratopsyche 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Ceratopsyche piatrix (Ross) 15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 15 Ceratopsyche morosa (Hagen) 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Hydroptilidae 0 1 3 0 0 0 1 0 0 0 0 0 0 0 0 5 Hydroptila 0 1 0 1 0 1 0 1 0 0 0 0 1 2 0 7

209 Oxyethira 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 2

Polycentropus 0 0 0 0 0 1 0 10 0 9 19 5 0 0 4 48

Cyrnellus fraternus (Banks) 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Glossosomatidae 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Agapetus 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Protoptila 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 2 Glossosoma intermedium 1 0 1 0 0 0 4 0 2 1 0 0 0 0 0 9 Lepidostoma 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 Oecetis 0 2 0 0 0 0 6 0 0 0 0 0 1 1 0 10 Nectopsyche 0 2 0 0 0 0 0 0 0 0 0 0 0 0 0 2 Triaenodes 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Mystacides sepulchralis (Walker) 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 3 Lepidoptera 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 2

Appendix G. Continued.

Ashley Creek Big Creek Sinking Creek Bay Creek Rocky Creek Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Psephenidae 6 0 2 0 0 0 0 0 0 0 0 0 0 0 0 8 Psephenus herricki (DeKay) 16 4 8 4 1 0 6 2 20 57 15 33 18 1 0 185 Ectopria 0 1 0 0 0 0 0 1 1 1 2 2 2 1 2 13 Hydrophilidae 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 Enochrus 0 6 0 0 0 0 0 0 0 0 0 0 0 0 0 6 Helichus 0 0 0 0 0 1 0 0 0 0 0 2 0 0 0 3 Tropisternus 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 Elmidae 18 11 6 1 0 0 28 1 24 5 5 2 5 0 15 121 Dubiraphia 0 0 1 2 17 0 0 16 0 0 1 1 0 8 21 67 Stenelmis 2 3 0 9 1 2 98 43 84 9 8 9 15 4 1 288 Optioservus 38 3 3 187 0 0 19 3 28 1 1 0 2 2 0 287 Gonielmis 0 1 3 0 12 7 0 0 1 0 0 0 0 0 0 24 Xenelmis 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 2 Stratiomys 0 0 3 0 0 0 0 0 0 0 0 0 0 0 0 3 Hydroporinae 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 210 Neoporus 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1

Diptera 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 2 Tipulidae 0 28 3 0 0 0 0 0 17 0 0 0 0 0 0 48 Antocha 1 0 1 1 0 0 1 0 0 0 0 0 0 0 0 4 Tipula 0 0 2 3 0 6 0 0 0 0 0 0 0 0 0 11 Hexatoma 0 0 2 0 0 6 31 0 113 0 0 0 13 5 0 170 Ceratopogonidae 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 2 Atrichopogon 0 2 0 0 0 1 0 0 0 0 1 12 4 0 1 21 Bezzia 1 11 2 2 1 4 2 0 0 0 0 0 4 15 2 44 Probezzia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 4 Stilobezzia 0 0 0 3 1 0 0 0 0 0 0 0 0 0 0 4 Mallochohelea 0 6 6 0 8 3 0 1 3 0 0 0 2 9 0 38 Culicoides 0 0 0 0 2 4 0 0 0 0 0 0 1 1 0 8

Appendix G. Continued.

Ashley Creek Big Creek Sinking Creek Bay Creek Rocky Creek Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Alluaudomyia 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 Empididae 2 0 1 0 0 0 2 0 1 0 0 0 0 0 0 6 Hemerodromia 7 3 5 8 1 2 13 5 17 3 5 1 2 0 0 72 Tanyderidae 0 0 0 0 2 0 0 0 0 0 0 0 1 0 0 3 Simuliidae 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 Simulium 6 0 0 3 0 0 0 0 20 4 1 23 9 0 0 66 Odontomyia 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Tabanus 0 0 0 0 0 0 0 0 0 0 0 0 0 3 1 4 Chironomidae 464 385 261 701 371 150 457 366 164 86 145 86 269 263 185 4353 Sciomyzidae 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Nemotelus 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 Pericoma 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1

211

Appendix H. Macroinvertebrates collected July 2013 from spring-fed tributary mesohabitats in the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation. Values are sums of original data from three subsamples.

Pulltite Spring Round Spring Alley Spring Blue Spring Cave Spring Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Platyhelminthes 5 2 1 2 1 5 13 0 3 4 6 12 7 2 2 65 Oligochaeta 3 84 19 51 33 226 23 29 35 6 53.5 19 42 37 117 778 Gordius 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Hirudinea 0 0 0 0 0 3 0 0 0 1 1 0 0 0 0 5 Ferrissia 0 0 0 0 0 0 0 0 0 0 0 0 0 1 2 3 Heliosoma 0 0 0 0 0 0 0 0 0 2 0 0 0 1 1 4 Physa 1 1 3 0 2 7 0 1 2 0 3 3 0 0 7 30 Elimia 4 0 135 22 0 0 0 0 0 15 62 8 31 2 10 289 Amnicola 193 205 28 69 0 3 99 2 23 433 223 587 36 0 4 1905 Fossaria 0 1 1 0 0 0 0 0 0 0 2 0 1 0 0 5 Corbicula 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Sphaerium 0 30 2 0 12 1 1 2 4 0 9 0 0 4 9 74 Hydracarina 14 4 25 105 0 1 17 4 18 47 30 32 6 2 1 306 Copepoda 0 0 0 0 0 2 0 0 0 0 3 0 0 1 1 7

Ostracoda 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0 2 212 Isopoda 0 0 0 0 0 0 0 0 0 0 0 0 0 5 2 7

Asellus 0 0 0 1 43 12 1 0 0 0 0 0 0 32 1 90 Lirceus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Orconectes 0 0 0 0 0 2 0 2 0 0 0 0 0 0 1 5 Gammarus 83 205 175 96 65 12 1 3 0 39 9 14 20 161 21 904 Gammarus pseudolimnaeus Fabricius 78 34 49 59 37 43 0 0 0 37 1 24 15 87 31 495 Gammarus fasciatus Say 0 0 0 0 0 0 2 1 3 0 0 0 0 0 0 6 Gammarus lacustrus Sars 0 0 0 0 0 0 0 0 0 0 0 0 3 0 0 3 Gammarus minus Say 0 0 0 0 0 0 0 0 0 0 1 2 0 0 0 3 Hyallela azteca (Saussure) 0 0 0 0 0 6 3 18 19 0 2 0 0 0 3 51 Collembola 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Hagenius brevistylus Selys 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Gomphus 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 2

Appendix H. Continued.

Pulltite Spring Round Spring Alley Spring Blue Spring Cave Spring Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Zygoptera 0 0 0 0 0 0 0 0 0 0 0 0 0 0 23 23 Calopterygidae 0 0 0 0 0 1 0 0 0 0 0 0 3 4 10 18 Leuctra 0 0 0 0 0 0 35 0 14 0 3 4 0 0 0 56 Perlodidae 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Ephemera 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Caenis 0 0 0 0 0 2 1 0 0 0 3 0 0 8 5 19 Tricorythodes 0 0 0 6 0 0 10 30 9 0 0 0 39 16 13 123 Heptageniidae 0 0 0 2 0 0 1 0 0 0 0 0 1 0 0 4 Stenacron 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Maccaffertium 0 0 0 0 0 0 0 0 0 0 0 0 5 1 0 6 Mecaffertium mediopunctatum McDunnough 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Stenonema femoratum (Say) 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Leucrocuta 0 0 0 4 0 0 1 0 0 0 0 0 0 0 0 5 Leptophlebia 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 2 Paraleptophlebia 0 0 0 0 0 0 0 4 1 0 0 0 0 0 0 5 Eurylophella 0 0 0 0 0 0 4 0 1 0 0 0 0 0 0 5

213 Serratella 0 0 0 0 0 0 6 0 0 0 1 0 8 0 0 15

Baetidae 0 0 0 5 0 0 10 0 1 46 1 21 3 0 3 90

Baetis 43 6 9 10 0 1 3 0 3 114 3 107 0 0 2 301 Centroptilum 0 0 0 1 0 12 0 0 2 0 0 1 0 1 0 17 Procloeon 0 0 0 1 0 1 0 0 0 0 1 1 0 0 0 4 Acentrella 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Plauditus 0 0 0 0 0 1 5 0 0 0 0 0 0 0 0 6 Acerpenna 0 0 0 0 0 0 0 0 0 0 0 0 2 2 14 18 Gerridae 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0 3 Trepobates 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Microvelia 0 0 1 0 0 0 0 0 0 0 0 1 0 0 1 3 Corixidae 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 2

Appendix H. Continued.

Pulltite Spring Round Spring Alley Spring Blue Spring Cave Spring Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Corydalidae 0 0 0 0 0 0 0 0 0 0 2 1 0 0 0 3 Sialis 0 1 0 0 0 0 0 1 0 0 2 0 0 2 0 6 Nigronia 0 1 1 1 0 0 4 0 0 0 0 0 0 0 0 7 Trichoptera 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Helicopsyche 0 0 0 2 0 0 1 0 0 0 0 0 2 0 2 7 Helicopsyche borealis (Hagen) 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Rhyacophilidae 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 Rhyacophila banksi Ross 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Lepidostoma 15 8 0 0 0 0 0 0 0 358 89 144 0 0 0 614 Psychomyia flavida Hagen 0 0 0 0 0 0 0 0 0 0 0 0 7 0 1 8 Cheumatopsyche 0 0 0 0 0 0 16 0 4 0 0 0 18 0 0 38 Ceratopsyche piatrix (Ross) 0 0 0 1 0 0 1 0 0 0 0 0 0 0 0 2 Ceratopsyche morosa (Hagen) 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 Hydroptilidae 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 2 Hydroptila 3 1 1 15 0 3 1 0 2 1 1 2 8 0 4 42 Polycentropus 0 0 0 0 0 0 5 3 0 0 0 0 0 0 1 9 214 Glossosomatidae 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1

Agapetus 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Protoptila 0 0 0 0 0 0 5 0 0 0 0 0 2 0 0 7 Glossosoma intermedium (Klapalek) 56 2 1 0 0 0 26 0 0 54 0 0 0 0 0 139 Lepidostoma 0 4 41 4 0 0 24 6 18 0 0 0 5 2 0 104 Brachycentrus lateralis (Say) 0 0 0 0 0 0 0 0 0 0 0 0 21 0 0 21 Micrasema 0 0 1 0 0 0 0 0 0 0 0 0 1 0 0 2 Leptoceridae 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 2 Nectopsyche 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 Triaenodes 0 0 0 0 0 2 0 1 0 0 0 0 2 0 1 6 Pycnopsyche 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 Psephenus herricki (DeKay) 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1

Appendix H. Continued.

Pulltite Spring Round Spring Alley Spring Blue Spring Cave Spring Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Peltodytes 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Helichus 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 Berosus 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Tropisternus 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Hydrobius 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Cymbiodyta 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Hydrobiomorpha 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Elmidae 0 0 0 2 0 0 1 0 0 0 0 1 1 0 0 5 Dubiraphia 0 0 0 0 0 0 0 0 0 0 0 0 0 5 3 8 Stenelmis 0 0 0 0 0 0 0 0 0 0 0 0 4 0 0 4 Optioservus 8 3 6 25 0 0 10 1 0 120 90 88 8 1 0 360 Heterelmis 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 Gonielmis 0 0 0 0 0 0 0 0 0 0 0 0 18 2 0 20 Dytiscidae 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Neoporus 0 0 0 0 0 6 0 0 0 0 0 0 0 0 0 6 Hydroporus 0 0 0 0 0 21 0 0 0 0 0 0 0 0 0 21

215 Enochrus 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1

Diptera 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1

Tipulidae 0 0 0 0 0 4 0 0 0 0 0 0 1 0 0 5 Antocha 0 0 0 0 0 0 0 0 0 6 0 1 0 0 0 7 Tipula 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Agabus 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 Cryptolabis 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Limnophila 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 Ceratopogonidae 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 Atrichopogon 0 0 0 0 0 6 0 0 0 0 1 0 0 0 2 9 Bezzia 0 0 1 0 0 8 1 2 0 2 1 1 0 1 3 20 Probezzia 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Appendix H. Continued.

Pulltite Spring Round Spring Alley Spring Blue Spring Cave Spring Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Stilobezzia 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 Mallochohelea 0 2 1 0 0 3 1 7 1 0 1 0 1 3 9 29 Culicoides 0 0 0 0 0 1 0 0 0 0 1 0 0 0 14 16 Alluaudomyia 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Hemerodromia 0 0 0 12 0 0 0 0 1 0 1 0 10 6 5 35 Chelifera 3 1 1 8 0 0 14 1 9 1 1 0 2 2 0 43 Clinocera 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 Dixa 0 0 1 0 0 0 1 0 0 0 2 0 0 0 0 4 Simulium 35 2 16 0 0 1 46 3 12 4 0 4 152 1 10 286 Gymnopais 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 Muscidae 0 0 1 0 0 0 2 0 0 2 1 2 0 0 0 8 Tabanus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 6 6 Chrysops 0 0 0 0 0 0 0 0 1 0 1 0 0 1 0 3 Anopheles 0 0 0 0 0 0 0 0 0 0 10 0 0 0 0 10 Chironomidae 42 307 123 491 103 90 154 305 386 50 246 258 306 167 185 3213 Sciomyzidae 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 216 Pericoma 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

Psychoda 0 0 0 0 0 4 0 0 0 0 0 0 0 0 0 4 Axymyiidae 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Pelecorhynchidae 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 Glutops 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 Phoridae 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1

Appendix I. Chironomidae collected in July and September 2013 from surface-fed tributary mesohabitats in the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation. Values are sums of original data from three subsamples.

Ashley Creek Bay Creek Big Creek Rocky Creek Sinking Creek Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Chironomidae 1 1 5 1 1 2 1 12 Tanypodinae 1 1 1 3 Ablabesmyia 2 7 8 4 9 7 16 53 Djalmabatista 3 1 4 Labrundinia 1 1 1 1 1 1 6 Larsia 1 1 Monopelopia 1 1 Natarsia Nilotanypus 6 5 1 6 1 1 1 21 Paramerina 0 Pentaneura 2 2 Procladius 1 1 3 1 6 Telopelopia 1 1 Thienemannimyia group 2 2 1 2 3 11 3 4 5 4 3 40 217 Zavrelimyia 2 2

Orthocladinae 3 1 2 6 Corynoneura 1 1 1 3 1 7 C/O group 4 1 1 21 1 3 4 35 Heterotrissocladius Parakiefferiellia 1 1 Parametriocnemus 3 2 2 1 1 1 1 11 Paraphaenocladius 1 1

Appendix I. Continued.

Ashley Creek Bay Creek Big Creek Rocky Creek Sinking Creek Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Rheocricotpus 4 2 1 1 1 9 Synorthocladius 3 1 4 Thienemanniella 1 1 1 1 4 Tvetenia 1 1 Xylotopus Chironominae 1 5 1 2 9 Chironomus 2 1 3 Cladotanytarsus 2 6 2 44 9 1 3 67 Cryptochironomus Cryptotendipes Dicrotendipes Glyptotendipes 1 1 Micropsectra 2 1 2 1 6 Microtendipes 1 5 2 8

218 Pagastiella 3 1 4

Paracladopelma 1 2 1 4

Paralauterborniella Paratanytarsus 1 1 1 1 1 1 2 8 Paratendipes Phaenopsectra 1 1 1 2 5 Polypedilum 18 2 1 1 4 1 5 18 4 40 6 3 103 Pseudochironomus 1 1 2 Rheotanytarsus 3 1 1 2 2 4 13

Appendix I. Continued.

Ashley Creek Bay Creek Big Creek Rocky Creek Sinking Creek Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Saetheria 1 1 Stempellinella 3 2 2 1 8 Stenochironomus 1 1 Strictochironomus 7 1 8 Sublettea 1 1 Tanytarsus 5 1 17 1 2 3 8 3 1 41 Tribelos 2 1 2 5

219

Appendix J. Chironomidae collected in July and September 2013 from spring-fed tributary mesohabitats in the Current and Jacks Fork rivers, Missouri. Veg = marginal vegetation. Values are sums of original data from three subsamples.

Alley Spring Blue Spring Cave Spring Pulltite Spring Round Spring Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Chironomidae 1 1 Tanypodinae 1 1 2 Ablabesmyia 1 1 1 3 Djalmabatista Labrundinia 1 1 Larsia 0 3 3 Monopelopia Natarsia 1 1 Nilotanypus Paramerina 1 2 3 Pentaneura Procladius 2 1 3 Telopelopia 220 Thienemannimyia group 2 2 2 6 9 1 3 3 4 1 1 34

Zavrelimyia 3 2 1 1 1 8 Orthocladinae 3 2 2 1 1 1 10 Corynoneura 1 1 2 C/O group 3 2 3 10 1 3 1 2 19 44 Heterotrissocladius 3 1 4 Parakiefferiellia Parametriocnemus 1 2 2 1 1 2 9 Paraphaenocladius

Appendix J. Continued.

Alley Spring Blue Spring Cave Spring Pulltite Spring Round Spring Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Rheocricotpus 2 10 1 1 1 15 Synorthocladius 1 4 5 Thienemanniella 2 4 1 7 Tvetenia 2 4 4 2 12 Xylotopus 1 1 Chironominae 1 1 1 2 5 Chironomus Cladotanytarsus Cryptochironomus 5 3 2 1 1 12 Cryptotendipes 1 1 Dicrotendipes 1 1 Glyptotendipes Micropsectra 2 1 3

Microtendipes 221 Pagastiella

Paracladopelma Paralauterborniella 2 2 Paratanytarsus Paratendipes 3 1 1 1 6 Phaenopsectra 2 1 2 5 Polypedilum 1 4 12 9 1 1 2 14 8 2 2 56 Pseudochironomus Rheotanytarsus 6 17 4 27

Appendix J. Continued.

Alley Spring Blue Spring Cave Spring Pulltite Spring Round Spring Taxa Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Riffle Pool Veg Total Saetheria Stempellinella 1 1 Stenochironomus Strictochironomus Sublettea Tanytarsus 5 2 4 2 1 4 1 1 1 1 22

Tribelos 1 1 2

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VITA

Rachel Heth graduated from Carthage High-School, Missouri in 2004. She began in the field of biology at Missouri Southern State University and earned a bachelor’s degree in 2008. During her time at Missouri Southern she had the opportunity to engage in an independent study and two National Science Foundation internships. It was her independent study on Plecoptera in Jones Creek (Missouri) with Robert Heth and her internship on the Ephemeroptera of lenthic habitats in Montana with Robert Newell that further confirmed her desire to focus in aquatic ecology. Rachel earned a master’s degree in biology from Missouri State University (MSU) in 2010 under the supervision of limnologist John Havel. At MSU she had the opportunity to work with the National Park

Service Heartland Inventory and Monitoring group in both the field and laboratory working on fish and macroinvertebrate collections. Experience with the Heartland team and further encouragement and guidance from David Bowles were critical steps in her decision to pursue a Ph.D. Rachel joined Robert Sites’ lab in the entomology department at the University of Missouri and completed her doctoral degree in 2015.

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