CLIMATE CHANGE – IMPACTS ON AQUATIC COMMUNITIES AND POTENTIAL MITIGATION VIA ALGAE BIOFUELS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1010429
• Project Start Date: Oct 31, 2016
• Project End Date: Sep 30, 2021
• Program Code: [(N/A)]- (N/A)

Recipient Organization
NEW MEXICO STATE UNIVERSITY
1620 STANDLEY DR ACADEMIC RESH A RM 110
LAS CRUCES,NM 88003-1239

Performing Department
Fisheries & Wildlife

Non Technical Summary
Climate change has been having drastic impacts on natural communities and has led to the extinction of many species. One example of a species that might be impacted by climate change is the Chihuahuan Mud Turtle (Kinosternon hirtipes). Presently it exists in only five ponds in south-west Texas.One way to slow climate change might be the production of algae biofuels. However, algae crashes can cause up to 30% of loss of crop production.Here, I propose to study baic life-history parameters of Chihuahuan Mud Turtle (Kinosternon hirtipes) and determine its outlook for survial. Furthermore, I will study mechanisms that can be used to reduce the probability of algae crashes in algae production ponds by mitigating invasions by undesired competitors (other algae) and predators (rotifers, ciliates, amoebas). Productivity and stability of algae polycultures will be compared to monocultures.

Animal Health Component

70%

Research Effort Categories

Basic

30%

Applied

70%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
136	0210	1070	100%

Knowledge Area
136 - Conservation of Biological Diversity;

Subject Of Investigation
0210 - Water resources;

Field Of Science
1070 - Ecology;

Keywords

chihuahuan mud turtle (kinsternon hirtipes)

algae cultivation

algae polycultures

Goals / Objectives
My long-term goals are twofold:Understand how climate-change driven environmental parameters effect aquatic populations and how those changes impacts the community as a whole.Contributing to making algae biofuel become a sustainable alternative to fossil fuels.My specific objectives for the next five years are:Uncover basic life-history parameters of the Chihuahuan Mud Turtle (Kinosternon hirtipes).Study mechanisms that can be used to reduce the probability of algae crashes in cultivation production ponds by studying mitigations of competitor and predator invasions.Compare productivity of algae polycultures to monocultures.Train graduate and undergraduate students in research to become thoughtful scientists and natural resource managers.

Project Methods
Here, I outline methods to study Chihuahuan Mud Turtle (Kinosternon hirtipes) and conduct algae experiments in the laboratory.Life-history of the Chihuahuan Mud Turtle (Kinosternon hirtipes)The Chihuahuan Mud Turtle (Kinosternon hirtipes) occurs from extreme west Texas, southwards into Chihuahua, Mexico, and south and east on the Mexican Plateau to the Chapala, Zapotlán, San Juanico, Pátzcuaro, and Valle de México basins (Iverson 1985; Ernst and Lovich 2009). In the United States, K. hirtipes is known only from the Alamito Creek drainage in Presidio County, Texas (Ernst and Lovich 2009).Turtles will be collected by traditional trapping methods using double throat wire mesh traps 1 m long x 30 cm diameter and sardines as bait. Traps will be completely submerged and checked every half hour to 45 minutes to keep the turtles from drowning. Each turtle will be returned to the water within 10 minutes of being captured to reduce stress on the animal. Since each turtle was given a unique identifier in a previous study (Cagle 1939), we will continue to collect data on individuals including growth measurements and movement between ponds.30 radio transmitters (manufacturer Holohil PD-2, http://holohil.com/bd2.htm) will be placed on the known populations in Presidio County and replaced the next year. This is approximately 18% of the population. The mean carapace length and weight for 250 individuals estimates the appropriate transmitter size at 3.8 g, and the pulse set at 220 will make the batteries last close to 12 months (Walde personal communication). Turtles will be tracked using two receivers by Titley Scientific and two H-antennae. Seasonal movement, overwintering habits and nesting areas and behaviors will be reported from these data (Wilson 1998, Peterson and Stone 2000, Row and Blouin-Demers 2006). In addition, the measurement data taken will be added to the existing 10 years of data gathered from our previous research and reported along with the telemetry findings.Algae ExperimentsWe will culture algae in BG 11 media. For the experiments, we will inoculate algae cultures at a starting optical density at 750 nm (OD750) of 0.04 in glass test tubes and place them into a rotator drum into an environmental chamber. For growth, daily optical density measurements will occur on the test tubes as a whole, using a Beckman CoulterTM DU® 530 life science UV/Vis spectrophotometer (Brea, CA) to eliminate the need to take samples, thus not reducing volume and risking contamination. Care will be taken to vortex and clean outer surface of tubes before measurement at 750, 680, and 450 nm.For HPLC extraction, we will vortex tubes using Fisher Scientific Vortex Mixer prior to sample extraction to homogenize algae solution. 200 µL samples will be taken from each tube on different days of the experiment, filtered, and stored in Corning® Spin-X® centrifuge tubes with the 0.22 µm nylon filter (Corning, NY) removed before storing at -80?C freezer in a secure container. Eppendorf microcentrifuge model 5424 will be used at 10,000 RPM for five minutes to filter samples completely before HPLC measurements.

Progress 10/01/20 to 09/30/21

Outputs
Target Audience:Endangered species managers, algae producers, scientific community. Changes/Problems:Unfortunately, our follow-up grant on increasing productivity of algae cultures was not funded and I will drop that aspect from my research focus. What opportunities for training and professional development has the project provided?In 2021, the project trained three graduate students in proposal and manuscript writing, presentation skills, and research. The project also trained 8 undergraduate students in research, creation of posters for presentations, and report writing. How have the results been disseminated to communities of interest?Via 5 peer-reviewed publications, 1 technical report,and 5 presentations. What do you plan to do during the next reporting period to accomplish the goals?For this coming year, I have a new proposal and the goals / specific objectives above will be revised. I will continue the success of native amphibians and reptiles in response to various selection pressures. I will also sample playa lakes of NM to test connectiveness and population dynamics of macroinvertebrates compared to populations in Kansas, North Dakota, Colorado, and east Texas. Results will be supplemented with experiments conducted in the field and the laboratory. I will also continue research on life-history parameters of endangered microsnail Pecos assiminea.

Impacts
What was accomplished under these goals? I graduated a non-thesis MSc student: Quijada, Bryanna.2021. Habitat preference of three New Mexican lizard species. NMSU, non-thesis Master's, 20 pp. I advised 3 graduate students and 8 undergraduate students in research projects. Together with students, I published 5 peer-reviewed papers and gave 3 presentations at Professional Meetings, and 2 presentations at local meetings. Regarding the specific objectives above, all questions have been answered and published in peer-reviewed journals and dozens of students have been trained over the last few years.

Publications

• Type: Journal Articles Status: Published Year Published: 2021 Citation: Canton, M.C., F.O. Holguin, C.C. Gard and W.J. Boeing. 2021. Allelochemical effect of gramine under temperature stress and impact on fat transesterification. Chemistry and Ecology 37: 481-492.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Guo, Z., W.J. Boeing, E. Borgomeo, Y. Xu and Y. Weng. 2021. Linking reservoir ecosystems research to the sustainable development goals. Science of the Total Environment 781: 146769.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Guo, Z., W.J. Boeing, Y. Xu, C. Yan, M. Faghihinia and D. Liu. 2021. Revisiting seasonal dynamics of total nitrogen in reservoirs with a systematic framework for mining data from existing publications. Water Research 201: 117380.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Oyebamiji, O.O., A.A. Corcoran, E. Navarro-Perez, M.O. Ilori, O.O. Amund, F.O. Holguin and W.J. Boeing. 2021. Lead tolerance and bioremoval by four strains of green algae from Nigerian fish ponds. Algal Research 58: 102403.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Guo, Z., W.J. Boeing, Y. Xu, E. Borgomeo, S.A. Mason and Y.G. Zhu. 2021. Global meta-analysis of microplastic contamination in reservoirs with a novel framework. Water Research 207: 117828.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Varela, M., L. Samaniego, W.J. Boeing. 2021. Diet analysis of native and invasive anurans in Sierra County, New Mexico. AZ/NM Joint Meeting of the Wildlife Society and American Fisheries Society, Virtual Conference.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Oyebamiji, O.O., M. Ilori, F. Holguin, W. Boeing. 2021. Lead tolerance and bioremoval by strains of green algae from a highly polluted Nigerian wastewater stream. Algal Biomass, Biofuels & Bioproducts, Virtual Conference.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Yarbrough, R., A. Foutch, O. Shackleton, C. Sidhe, W.J. Boeing. 2021. Cryptic micro-snail Assiminea pecos - survey methods and habitat. 66th Annual New Mexico Water Conference, Virtual Conference (poster).

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:Endangered species managers, algae producers, scientific community. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project trained four graduate students in proposal and manuscript writing, presentation skills, and research. The project also trained one undergraduate student in research and report writing. How have the results been disseminated to communities of interest?Via one peer-reviewed publication, one technical report, one popular report, and 3 presentations. What do you plan to do during the next reporting period to accomplish the goals?The next reporting period will focus on dynamics of large invertebrates (branchiopods) in playa lakes. Additionally, we will research survey methods of an endangered snail, and start a reptile project at the Jornada LTER.

Impacts
What was accomplished under these goals? 1. A MSc student finished her thesis looking at how frog populations respond to biotic and abiotic factors. 2. A PhD student finished his dissertation studying how algae turf scrubbers (ATS) could be efficient for production of algae biomass as well as aquaculture products (e.g., shrimp). We published 1 peer-reviewed paper and he gave three presentations.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Siville, B.* and W.J. Boeing#. 2020. Optimization of algal turf scrubber (ATS) technology through targeted harvest rate. Bioresource Technology Reports 9: 100360.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Hatch M.C., Abadi F., Boeing W.J., Sabela L., Porter M.D., Cowley D.E. 2020. Sustainability management of short-lived freshwater fish in human-altered ecosystems should focus on adult survival. PloS ONE 15: e0232872
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Siville, B., W.J. Boeing (2020). Algae Consortia and Microbiomes- Improving Culture Health and Performance through Diversity: Patch Mosaic Paradigms in Algal Turf Scrubber Substrate. Oral presentation, Algae Biomass Summit, online.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Siville, B., W.J. Boeing (2020). Patch Mosaic Paradigms in Algal Turf Scrubber Substrate. Poster, Algae Biomass Summit, online.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Siville, B., W.J. Boeing (2020). Patch Mosaic Paradigms in Algal Turf Scrubber Substrate. Poster, 65th New Mexico Water Conference, online.
• Type: Theses/Dissertations Status: Published Year Published: 2020 Citation: Siville, Bert. 2020. ATS Technology: Function and optimization in small scale aquaculture. NMSU Dissertation, 99pp.
• Type: Theses/Dissertations Status: Published Year Published: 2020 Citation: Samaniego, Lauren. 2020. Synthesizing biotic and abiotic factors responsible for the success of native and alien ranids on a ranch in southern New Mexico. NMSU Thesis, 72 pp.

Progress 10/01/18 to 09/30/19

Outputs
Target Audience:Thetarget audiences are scientists, managers, decision-makers, and the general community. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?There have been seven papers published in peer-reviewed journals, a book chapter, three popular articles, and gave nine presentations at professional conferences. What do you plan to do during the next reporting period to accomplish the goals?The focus will be on publishing additional findings on aquatic foodwebs and usefulness of algae fpr the future of energy generation, environmental mitigation, and sustainability on human food consumption

Impacts
What was accomplished under these goals? In terms of specific objectives: 1. Research hasuncovered basic life-history paramters of Kinosternon hirtipes and two papers have been publishedon nesting behavior and predation behaviors. 2. Four papers were publishedin 2019 on the usefulness of algae (biodetoxification of heavy metals), increased stability through allelopathic interactions, and investigated algal turf scrubber technology as an alternative method for growing algae.

Publications

Progress 10/01/17 to 09/30/18

Outputs
Target Audience:My target audiences are scientists, managers, decision makers, and the general community. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?In 2019, I directly supervised 5 graduate students and 5 undergraduate students. How have the results been disseminated to communities of interest?We published seven papers in peer-reviewed journal, a book chapter, three popular articles, and gave nine presentations at professional conferences. What do you plan to do during the next reporting period to accomplish the goals?The focus will be on publishing additional findings on aquatic foodwebs and usefulness of algae fpr the future of energy generation, environmental mitigation, and sustainability on human food consumption.

Impacts
What was accomplished under these goals? In terms of specific objectives: 1. We uncovered basic life-history paramters of Kinosternon hirtipes and published 2 papers on mesting behavior and predation behaviors. 2. We published 4 papers in 2019 on usefullness of algae (biodetoxification of heavy metals), increased stability through allelopathic interactions, and investigated algal turf scrubber technology as an alternative method for growing algae.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Siville, B. and W.J. Boeing. 2020. Optimization of algal turf scrubber (ATS) technology through targeted harvest rate. Bioresource Technology Reports 9: 100360.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Campos, H., W.J. Boeing, H.L. Throop. 2019. Decaying woodrat (Neotoma spp.) middens increase soil resources and accelerate decomposition of contemporary litter. Journal of Arid Environments 170: 104007.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Canton, M.C., F.O. Holguin, W.J. Boeing. 2019. Alkaloid gramine to control algal invaders: Algae inhibition and gramine persistence. Bioresource Technology Reports 7: 100304.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Oyebamiji, O.O., W.J. Boeing, F.O. Holguin, O. Ilori, O. Amund. 2019. Green microalgae cultured in textile wastewater for biomass generation and biodetoxification of heavy metals and chromogenic substances. Bioresource Technology Reports 7: 100247.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Corcoran, A.A., M. Seger, R. Niu, N. Nirmalakhandan, P.J. Lammers, F.O. Holguin, W.J. Boeing. 2019. Evidence of induced allelopathy in an isolate of Coelastrella following co-culture with Chlorella sorokiniana. Algal Research 41: 101535.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Platt, S.G., J.L. Smith, T.R. Rainwater, W.J. Boeing. 2019. Notes on the predation of rough-footed mud turtles (Kinosternon hirtipes) in West Texas, USA. Western North American Naturalist 79: 130-134.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Smith, J.L., S.G. Platt, R.L. Martin, W.J. Boeing. 2018. Kinosternon hirtipes (rough-footed mud turtle) nesting behavior and descriptions of nest sites. Herpetological Review 49: 109-110.
• Type: Book Chapters Status: Published Year Published: 2019 Citation: Boeing, W.J. 2019. Defensive Avoidance. In: Choe, J.C. (Ed.), Encyclopedia of Animal Behavior, 4: 292-297.
• Type: Other Status: Published Year Published: 2019 Citation: Boeing, W.J. 2019. GALS, a personal account. AAUW NM Fall Roadrunner.
• Type: Other Status: Published Year Published: 2019 Citation: Boeing, W.J. 2019. Exploring the Gila Wilderness with Girls on outdoor Adventures for Leadership and Science. USDA Forest Service News.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Samaniego, L., W.J. Boeing. 2019. Occupancy of native leopard frogs in response to biotic and abiotic variables. Chihuahuan Desert Conference, El Paso, TX, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Oyebamiji, O., W.J. Boeing, F.O. Holguin, P. Cooke, O. Amund, M. Ilori. 2018. Antibiotic stress and light intensity: two major factors involved in the accumulation of carotenoid pigments in green microalgae isolates. Bioresource Technology for Bioenergy, Bioproducts, and Environmental Sustainability, Sitges, Spain.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Oyebamiji, O., W.J. Boeing, F.O. Holguin, O. Amund, M. Ilori. 2018. Green microalgae species cultured in textile wastewater for biomass generation and biodetoxification of heavy metals and chromogenic substances. Bioresource Technology for Bioenergy, Bioproducts and Environmental Sustainability, Sitges, Spain.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Galarza, J., E. Delgado, S. Munson-McGee, W.J. Boeing, D.J. Valles-Rosales. 2018. Glandless cottonseed meal as a protein source for aquaculture. American Society of Agricultural and Biological Engineers, Las Cruces, NM, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Varela, M., L.A. Samaniego, W.J. Boeing. 2019. Diet analysis of native and non-native ranid species in Sierra County, New Mexico. Chihuahuan Desert Conference, El Paso, TX, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Boeing, W.J., K. Swaim, N. Macanowicz, S. Hylander, C. Koebsch. 2019. From bacteria to fish - what spins the aquatic food web in desert sinkholes? Chihuahuan Desert Conference, El Paso, TX, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Samaniego, L., W.J. Boeing. 2019. Occupancy of American Bullfrog (Lithobates catesbeianus) and ecosystem impacts. AZ/NM Joint Meeting of the Wildlife Society and American Fisheries Society, Albuquerque, NM, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Siville, B., W.J. Boeing. 2019. Remediation of aquatic ecosystems with algae turf scrubbers (ATS) - Optimization. AZ/NM Joint Meeting of the Wildlife Society and American Fisheries Society, Albuquerque, NM, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Flynn, L.M., D. Doublet, L. Kremer, T. Rossettie, W.J. Boeing. 2019. Challenges and rewards of starting a teenage mentorship program in a university STEM department. AZ/NM Joint Meeting of the Wildlife Society and American Fisheries Society, Albuquerque, NM, USA.

Progress 10/31/16 to 09/30/17

Outputs
Target Audience:My target audience is other scientists as well as managers and decision makers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Currently, I am supervising 2 PhD students studentsand1 undergraduate students in research project in my laboratory. How have the results been disseminated to communities of interest?We continue to publish results in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?We will conduct experiments looking at various harvest rates of filamentous algae on Algae Turf Scrubbers and implement turf scrubbers into a wastewater stream and study nutrient and metal uptake. Several experiments regarding valuable products and benefits from various pelagic algae have already been conducted are ongoing.

Impacts
What was accomplished under these goals? We were able to further our understanding towards my long-term goals as follows: 1. Using data from a decade-long mark-recapture effort, we discovered that the US population of the rough-footed mud turtle (Kinosternon hirtipes) consists of only 134 +- 52 individuals at this point and is highly imperiled. In 1986, 9 aquatic habitats existed for this turtle species. And while one habitat was lost due to draught conditions, 5 were drained by landowners, leaving only 3 habitats. Thus, while climate change does impact this aquatic turtle population, the direct threat of human impact far outweights climate issues at this point. Furthermore, we report various ecological (population is skewed towards males and males are larger in size) and habitat (turtles thrive in pond with abundance of underwater logs and they brumate in close proximity to aquatic habitat) information for K. hirtipes to aid in conservation efforts for the future. 2. To make algae biofuel a viable enterprise, the research has to focus on (a) increasing productivity, (b) finding benefits other than biofuel for algae cultivation, and (c) easier harvesting methods. We have successfully installed algae turf scrubber (ATS) in aquaria in the laboratory, that might be able to aid will all three issues mentioned above. ATS support filiamentous algae growing on a substrate that also have the ability to filter the water flowing across them. Currently, we are focusing our efforts on the following research questions: 1) Can we increase algae biomass using algae turf scrubbers (ATS) by optimizing harvest rate? 2) Can ATS be used to effectively eliminate nutrients and metals from wastewater and irrigation water (that is being returned to the river)? 3) Are there any other valuable co-products or benefits (e.g., antibiotic properties)that can be harvested from various algae strains? 4) What life-history stage is most important to protect to guarantee survival of endangered turtle Kinosternon hirtipes.

Publications

• Type: Journal Articles Status: Published Year Published: 2017 Citation: Lammers, P.J., M. Huesemann, W. Boeing, D.B. Anderson, R.G. Arnold, X. Bai, K. Balachandran, M. Bohle, Y. Brhanavan, L. Brown, J. Brown, J.K. Brown, S. Chisholm, C.M. Downes, S. Fulbright, Y. Ge, J.E. Holladay, A. Khopkar, A. Koushik, P. Laur, B.L. Marrone, J.B. Mott, N. Nirmalakhandan, K.L. Ogden, R.L. Parsons, J. Polle, R.D. Ryan, T. Samocha, M. Seger, T. Selvaratnam, R. Sui, A. Thomasson, A. Unc, W. Van Voorhies, P. Waller, Y. Yao, J. Olivares. 2017. Review of the cultivation program within the National Alliance for Advanced Biofuels and Bioproducts. Algal Research 22: 166-186.
• Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Smith, J.L., S.G. Platt, W.J. Boeing. KINOSTERNON HIRT/PBS (Rough-footed Mud Turtle). Oveerwintering Habitats. Herpetological Reviews
• Type: Journal Articles Status: Submitted Year Published: 2018 Citation: Koebsch, C. and W.J. Boeing. Bacterial beta-glucosidase activity across large environmental gradients in desert sinkholes is dominated by abiotic factors. Aquatic Microbial Ecology
• Type: Other Status: Other Year Published: 2017 Citation: Boeing, W.J. 2017. From Bacteria to Fish: What spins the Aquatic Food Web in Desert Sinkholes? Invited Seminar. University of Ningbo, China.