Effects of Earthworms on Post-Industrial Polluted Soil Remediation
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©2017 Nicholas J. Henshue ALL RIGHTS RESERVED EFFECTS OF EARTHWORMS ON POST-INDUSTRIAL POLLUTED SOIL REMEDIATION by NICHOLAS J. HENSHUE A dissertation submitted to the School of Graduate Studies Rutgers, The State University of New Jersey In partial fulfillment of the requirements For the degree of Doctor of Philosophy Graduate Program in Ecology and Evolution Written under the direction of Claus Holzapfel And approved by: _____________________________________ _____________________________________ _____________________________________ _____________________________________ New Brunswick, New Jersey October, 2017 ABSTRACT OF THE DISSERTATION Effects of Earthworms on Post-Industrial Polluted Soil Remediation by By NICHOLAS J HENSHUE Dissertation Director: Claus Holzapfel This dissertation consists of a literature review and three main chapters, all of which are central to assessing the impacts of earthworms in polluted metalliferous soils. Chapter One gives a background in earthworm ecology, then summarizes the questions asked for this study, and gives a brief literature review of the work that has been done prior to this research. Chapter Two describes a biological survey completed through three sampling seasons, which indicate species prevalence and population density in both non-polluted fields and forests, and metal-contaminated brownfields. This survey used a standardized mustard water extraction protocol over 135 sites in Pennsylvania and New Jersey, USA. Although there was a great deal of variation in numbers across sites, both polluted and non-polluted areas contained primarily 3 species– all of which are non-native. A large community pot experiment is the topic of Chapter Three, where two types of plants and four earthworm species were placed in three different soil controls of low, ii mid, and most metal pollution loading and grown together in five-gallon (19 L) buckets for 10 weeks. The primary goals of this experiment were to assess earthworm and plant communities and their potential interactions regarding growth and metal uptake in containers with and without earthworm communities. After all plants were harvested, they were analyzed for metal content including arsenic (As), cadmium (Cd), lead (Pb), copper (Cu), and zinc (Zn). Chapter Four details a population level experiment, examining individual plants in the pots. This design removes the competition coefficient from having two different plant species together in a community-structured experiment like the previous chapter. This design used 1-gallon tall tree sapling pots with the same soil treatments (low, mid, most pollution levels) as chapter three. This experiment, however, only used one type of plant per pot, and only two species of earthworm; both of which have been very common in all sites studied for the biological survey detailed in Chapter Two. After 60 days of growth, the plants, soil and earthworms were harvested and analyzed for metal content of arsenic, chromium, copper, lead, and zinc. iii ACKNOWLEDGEMENTS: I wish to wholeheartedly thank my committee members: The outstanding Claus Holzapfel, Frank Gallagher, Peter Groffman, and Mark Hodson for all of their mentorship and support. Thanks to the incredible department administrator Marsha Morin for making sure all the paperwork was perfect. Thank you to the wonderful scientists at the Cary Institute of Ecosystem Studies, especially Denise Schmidt and Milada Vomela. None of the analysis would have been possible without you two. Thanks to my fellow worm-wrangler and friend, Kathleen Farley, and a huge acknowledgement to those young researchers that helped along the way: aspiring scientists like Rita Matos, Omar Abukwaik, and Zain Abidin who collected thousands of earthworms from all over New Jersey and Kyle Froehlich, who helped me define exact characteristics of my study plants. A special thank you to the Rutgers Ecology and Evolution cohort of 2012 (or was it 2013?) for their love and support— When we keep each other up, we can all go higher. A huge amount of thanks and love to my family, Deb Henshue, Tom Henshue, and Steph Henshue, for the support; whether it be babysitting, moral, or financial, you were always there. And most importantly, my beloved wife, Jen. She has caught earthworms, given up her space, time, and husband for months at a time, and endured specimens in the refrigerator far more than any person should have. While I may be a Doctor, you will always have more patience. I love you forever. iv Table of Contents: Abstract ...................................................................................... ii Acknowledgements ........................................................................... iv Chapter 1: Introduction to the Ecology of Earthworms in the Northeastern United States ............................................ 1 I. Abstract ............................................................................................... 1 II. Introduction to Earthworm Ecology in North America ..................... 3 A. Anecic Earthworms ........................................................................ 3 B. Endogeic Earthworms .................................................................... 5 C. Epigeic Earthworms ....................................................................... 5 D. Epi-endogeic Earthworms .............................................................. 6 E. Native, Non-Native Earthworms ................................................... 7 III. Earthworms Affect Forests ................................................................... 9 IV. Earthworms in Contaminated Soil ...................................................... 11 V. Outline of Chapters and Synopsis of Issues ........................................ 14 VI. References ............................................................................................ 17 Chapter 2: Biological Survey of Polluted and Non-polluted Soils Across a Longitudinal Gradient ...................................... 23 I. Abstract ................................................................................................ 23 II. Introduction ......................................................................................... 24 III. Sampling Methods ............................................................................... 27 A. Hand Sorting ................................................................................. 28 B. Formalin ......................................................................................... 29 C. Electrical Octet Method ................................................................ 30 D. Pure Allyl Isothiocyanate ............................................................... 31 E. Mustard and Mustard Powder ....................................................... 32 F. Method Review and Conclusion ................................................... 33 IV. Sampling Locations .............................................................................. 34 v V. Methods ............................................................................................... 36 VI. Results .................................................................................................. 40 A. Temporal and Biomass Distribution ............................................. 41 B. Specimen Counts in Non-polluted and Polluted Sites ................. 42 C. Analysis of Species ......................................................................... 45 D. Analysis of Diversity Indices .......................................................... 46 E. Canonical Correspondence Analysis of Assessment Results ........ 48 VII. Discussion ............................................................................................ 52 VIII. Conclusion ........................................................................................... 54 IX. References ............................................................................................ 56 X. Appendix .............................................................................................. 63 A. Map of All Biosurvey Sampling Sites (Large) ................................ 63 B. Large CCA of Biplot Results ......................................................... 64 C. Complete Chart of Earthworm Biosurvey Results ........................ 65 Chapter 3: Earthworm Impacts on Phytoremediation in Metalliferous Soil Communities ..................................... 70 I. Abstract ................................................................................................ 70 II. Introduction ......................................................................................... 72 III. Site History ........................................................................................... 74 IV. Soil Characterization ........................................................................... 77 V. Plant Selection ..................................................................................... 78 VI. Earthworm Selection ........................................................................... 79 VII. Methods ............................................................................................... 80 VIII. Results .................................................................................................. 86 IX. Discussion ............................................................................................ 95 X. Conclusion ..........................................................................................