Skip to main content

Advertisement

SpringerLink
Log in

Highlighting the promise of qPCR-based environmental monitoring: response of the ribosomal RNA:DNA ratio of calanoid copepods to toxic cyanobacteria

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

Calanoid copepods are integral to aquatic food webs and may drive the bioaccumulation of toxins and heavy metals, spread of infectious diseases, and occurrence of toxic cyanobacterial harmful algal blooms (HABs) in freshwater aquatic systems. However, interrelationships between copepod and cyanobacterial population dynamics and ecophysiology remain unclear. Insights into these relationships are important to aquatic resource management, as they may help guide mitigation efforts. We developed a calanoid copepod qPCR assay to investigate how copepod abundance and physiological status relate to the abundance of cyanobacteria and the concentration of total microcystin in a HAB-prone freshwater multi-use eutrophic lake. Through in silico and in vitro validation of primers and analyses of time series, we demonstrate that our assay can be used as a reliable tool for environmental monitoring. Importantly, copepod RNA:DNA ratios on and shortly after the day when microcystin concentration was at its highest within the lake were not significantly lower (or higher) than before or after this period, suggesting that copepods may have been tolerant of microcystin levels observed and capable of perpetuating bloom events by consuming competitors of toxic cyanobacteria.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

The data used in this study will be publicly accessible in the Sci-Hub repository upon publication of the study.

References

  • Anderson MJ (2001) A new method for non-parametric multivariate analysis of variance. Austral Ecol 26:32–46

    Google Scholar 

  • Aw TG, Rose JB (2012) Detection of pathogens in water: from phylochips to qPCR to pyrosequencing. Curr Opin Biotechnol 23:422–430

    Article  CAS  Google Scholar 

  • Banerji A, Bagley MJ, Elk M, Pilgrim E, Martinson J, Santo Domingo JW (2018) Spatial and temporal dynamics of a freshwater eukaryotic plankton community revealed via 18S rRNA gene metabarcoding. Hydrobiologia 818:71–86

    Article  CAS  Google Scholar 

  • Banerji A, Bagley MJ, Shoemaker JA, Tettenhorst DR, Nietch CT, Allen HJ, Santo Domingo JW (2019) Evaluating putative ecological drivers of microcystin spatiotemporal dynamics using metabarcoding and environmental data. Harmful Algae 86:84–95

    Article  CAS  Google Scholar 

  • Bayly IE (1963) Parasitic castration of a freshwater calanoid copepod by a cestode cysticercoid stage. Crustaceana 5:75–80

    Article  Google Scholar 

  • Benoît HP, Johannsson OE, Warner DM, Sprules WG, Rudstam LG (2002) Assessing the impact of a recent predatory invader: the population dynamics, vertical distribution, and potential prey of Cercopagis pengoi in Lake Ontario. Limnol Oceanogr 47:626–635

    Article  Google Scholar 

  • Berry JP, Jaja-Chimedza A, Dávalos-Lind L, Lind O (2012) Apparent bioaccumulation of cylindrospermopsin and paralytic shellfish toxins by finfish in Lake Catemaco (Veracruz, Mexico). Food Addit Contam Part A 29:314–321

    Article  CAS  Google Scholar 

  • Boon PI, Bunn SE, Green JD, Shiel RJ (1994) Consumption of cyanobacteria by freshwater zooplankton: implications for the success of ‘top-down' control of cyanobacterial blooms in Australia. Aust J Mar Freshw Res 45:875–887

    Article  Google Scholar 

  • Burns CW, Schallenberg M (2001) Calanoid copepods versus cladocerans: consumer effects on protozoa in lakes of different trophic status. Limnol Oceanogr 46:1558–1565

    Article  CAS  Google Scholar 

  • Calliari D, Andersen CM, Thor P, Gorokhova E, Tiselius P (2006) Salinity modulates the energy balance and reproductive success of co-occurring copepods Acartia tonsa and A. clausi in different ways. Mar Ecol Prog Ser 312:177–188

    Article  Google Scholar 

  • Cannon MV, Hester J, Shalkhauser A, Chan ER, Logue K, Small ST, Serre D (2016) In silico assessment of primers for eDNA studies using PrimerTree and application to characterize the biodiversity surrounding the Cuyahoga River. Sci Rep 6:22908

    Article  CAS  Google Scholar 

  • Carter JCH (1969) Life cycles of Limnocalanus macrurus and Senecella calanoides, and seasonal abundance and vertical distributions of various planktonic copepods, in Parry Sound, Georgian Bay. J Fish Res Board Can 26:2543–2560

    Article  Google Scholar 

  • Flinkman J, Vuorinen I, Christiansen M (1994) Calanoid copepod eggs survive passage through fish digestive tracts. ICES J Mar Sci 51:127–129

    Article  Google Scholar 

  • Gopalan G, Culver DA, Wu L, Trauben BK (1998) Effects of recent ecosystem changes on the recruitment of young-of-the-year fish in western Lake Erie. Can J Fish Aquat Sci 55:2572–2579

    Article  Google Scholar 

  • Hansson L-A, Gustafsson S, Rengefors K, Bomark L (2007) Cyanobacterial chemical warfare affects zooplankton community composition. Freshw Biol 52:1290–1301

    Article  CAS  Google Scholar 

  • Holmborn T, Gorokhova E (2008) Relationships between RNA content and egg production rate in Acartia bifilosa (Copepoda, Calanoida) of different spatial and temporal origin. Mar Biol 153:483–491

    Article  CAS  Google Scholar 

  • Jamieson CD (2005) Coexistence of two similar copepod species, Eudiaptomus gracilis and E. graciloides: the role of differential predator avoidance. Hydrobiologia 542:191–202

    Article  Google Scholar 

  • Jonsson A, Nielsen TG, Hrubenja I, Maar M, Petersen JK (2009) Eating your competitor: functional triangle between turbulence, copepod escape behavior and predation from mussels. Mar Ecol Prog Ser 376:143–151

    Article  CAS  Google Scholar 

  • Jungbluth MJ, Goetze E, Lenz PH (2013) Measuring copepod naupliar abundance in a subtropical bay using quantitative PCR. Mar Biol 160:3125–3141

    Article  Google Scholar 

  • Kadiene EU, Bialais C, Ouddane B, Hwang J-S, Souissi S (2017) Differences in lethal response between male and female calanoid copepods and life cycle traits to cadmium toxicity. Ecotoxicology 26:1227–1239

    Article  CAS  Google Scholar 

  • Kadiene EU, Ouddane B, Hwang J-S, Souissi S (2019) Bioaccumulation of metals in calanoid copepods by oral intake. Sci Rep 9:9492

    Article  Google Scholar 

  • Kang Y, Koch F, Gobler CJ (2015) The interactive roles of nutrient loading and zooplankton grazing in facilitating the expansion of harmful algal blooms caused by the pelagophyte, Aureoumbra lagunensis, to the Indian River Lagoon, FL, USA. Harmful Algae 49:162–173

    Article  CAS  Google Scholar 

  • Kapoor V, Elk M, Li X, Impellitteri CA, Santo Domingo JW (2016) Effects of Cr (III) and Cr (VI) on nitrification inhibition as determined by SOUR, function-specific gene expression and 16S rRNA sequence analysis of wastewater nitrifying enrichments. Chemosphere 147:361–367

    Article  CAS  Google Scholar 

  • Kapoor V, Pitkänen T, Ryu H, Elk M, Wendell D, Santo Domingo JW (2015) Distribution of human-specific bacteroidales and fecal indicator bacteria in an urban watershed impacted by sewage pollution, determined using RNA- and DNA-based quantitative PCR assays. Appl Environ Microbiol 81:91–99

    Article  Google Scholar 

  • Kiørboe T, Saiz E, Viitasalo M (1996) Prey switching behaviour in the planktonic copepod Acartia tonsa. Mar Ecol Prog Ser 143:65–75

    Article  Google Scholar 

  • Koski M, Schmidt K, Engström-Öst J, Viitasalo M, Jónasdóttir S, Repka S, Sivonen K (2002) Calanoid copepods feed and produce eggs in the presence of toxic cyanobacteria Nodularia spumigena. Limnol Oceanogr 47:878–885

    Article  Google Scholar 

  • LeBlanc F, Belliveau V, Watson E, Coomber C, Simard N, DiBacco C, Bernier R, Gagné N (2020) Environmental DNA (eDNA) detection of marine aquatic invasive species (AIS) in Eastern Canada using a targeted species-specific qPCR approach. Manag Biol Invasions 11:201–217

    Article  Google Scholar 

  • Liebig R, Vanderploeg HA (1995) Vulnerability of Dreissena polymorpha larvae to predation by Great Lakes calanoid copepods: the importance of the bivalve shell. J Great Lakes Res 21:353–358

    Article  Google Scholar 

  • Magnusson K, Tiselius P (2010) The importance of uptake from food for the bioaccumulation of PCB and PBDE in the marine planktonic copepod Acartia clausi. Aquat Toxicol 98:374–380

    Article  CAS  Google Scholar 

  • McArdle BH, Anderson MJ (2001) Fitting multivariate models to community data: a comment on distance-based redundancy analysis. Ecology 82:290–297

    Article  Google Scholar 

  • Meerhoff M, Iglesias C, de Mello FT, Clemente JM, Jensen E, Lauridsen TL, Jeppesen E (2007) Effects of habitat complexity on community structure and predator avoidance behaviour of littoral zooplankton in temperate versus subtropical shallow lakes. Freshw Biol 52:1009–1021

    Article  Google Scholar 

  • Miralto A, Barone G, Romano G, Poulet SA, Ianora A, Russo GL, Buttino I, Mazzarella G, Laabir M, Cabrini M, Giacobbe MG (1999) The insidious effect of diatoms on copepod reproduction. Nature 402:173–176

    Article  CAS  Google Scholar 

  • Mitra A, Flynn KJ (2006) Promotion of harmful algal blooms by zooplankton predatory activity. Biol Lett 2:194–197

    Article  Google Scholar 

  • Ning J, Li C, Yang G, Wan A, Sun S (2013) Use of RNA:DNA ratios to evaluate the condition and growth of the copepod Calanus sinicus in the Southern Yellow Sea. Deep Sea Res Part II Top Stud Oceanogr 97:109–116

    Article  CAS  Google Scholar 

  • Paerl HW, Gardner WS, Havens KE, Joyner AR, McCarthy MJ, Newell SE, Qin B, Scott JT (2016) Mitigating cyanobacterial harmful algal blooms in aquatic ecosystems impacted by climate change and anthropogenic nutrients. Harmful Algae 54:213–222

    Article  Google Scholar 

  • Piasecki W, Goodwin AE, Eiras JC, Nowak BF (2004) Importance of Copepoda in freshwater aquaculture. Zool Stud 43:193–205

    Google Scholar 

  • Rasch EM, Wyngaard GA (2006) Genome sizes of cyclopoid copepods (Crustacea): evidence of evolutionary constraint. Biol J Linn Soc 87:625–635

    Article  Google Scholar 

  • Reichwaldt ES, Song H, Ghadouani A (2013) Effects of the distribution of a toxic Microcystis bloom on the small scale patchiness of zooplankton. PLoS One 8:e66674

    Article  CAS  Google Scholar 

  • Ryu H, Henson M, Elk M, Toledo-Hernandez C, Griffith J, Blackwood D, Noble R, Gourmelon M, Glassmeyer S, Santo Domingo JW (2013) Development of quantitative PCR assays targeting the 16S rRNA genes of Enterococcus spp. and their application to the identification of Enterococcus species in environmental samples. Appl Environ Microbiol 79:196–204

    Article  CAS  Google Scholar 

  • Saiz E, Calbet A, Fara A, Berdalet E (1998) RNA content of copepods as a tool for determining adult growth rates in the field. Limnol Oceanogr 43:465–470

    Article  CAS  Google Scholar 

  • Schaus MH, Vanni MJ (2000) Effects of gizzard shad on phytoplankton and nutrient dynamics: role of sediment feeding and fish size. Ecology 81:1701–1719

    Article  Google Scholar 

  • Smith AS, Alexander Jr JE (2008) Potential effects of the freshwater jellyfish Craspedacusta sowerbii on zooplankton community abundance. J Plankton Res 30:1323–1327

    Article  Google Scholar 

  • Sommer U, Sommer F (2006) Cladocerans versus copepods: the cause of contrasting top–down controls on freshwater and marine phytoplankton. Oecologia 147:183–194

    Article  Google Scholar 

  • Speekmann CL, Nunez BS, Buskey EJ (2007) Measuring RNA:DNA ratios in individual Acartia tonsa (Copepoda). Mar Biol 151:759–766

    Article  CAS  Google Scholar 

  • Stemberger RS, Evans MS (1984) Rotifer seasonal succession and copepod predation in Lake Michigan. J Gt. Lakes Res 10:417–428

    Article  Google Scholar 

  • Wagner M, Durbin E, Buckley L (1998) RNA:DNA ratios as indicators of nutritional condition in the copepod Calanus finmarchicus. Mar Ecol Prog Ser 162:173–181

    Article  CAS  Google Scholar 

  • White MM, McLaren IA (2000) Copepod development rates in relation to genome size and 18S rDNA copy number. Genome 43:750–755

    Article  CAS  Google Scholar 

  • Wyngaard GA, McLaren IA, White MM, Sévigny JM (1995) Unusually high numbers of ribosomal RNA genes in copepods (Arthropoda: Crustacea) and their relationship to genome size. Genome 38:97–104

    Article  CAS  Google Scholar 

  • Ye J, Coulouris G, Zaretskaya I, Cutcutache I, Rozen S, Madden T (2012) Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinform 13:134

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We would like to thank Megan Berberich for technical assistance with laboratory procedures; Joel Allen, Mia Vijanderan, Dana Macke, Kit Daniels, and Armah de la Cruz for their role in collecting, transporting, and processing the lake water samples used in this study; Jade Young (Army Corps of Engineers) for sharing 2016 calanoid densities for Lake Harsha; Chris Nietch and Stefania Guglielmi for helping to validate discrete sonde water quality measurements; and John Olszewski and Samuel Hayes for critically reviewing an earlier draft of this manuscript.

Funding

The U.S. Environmental Protection Agency, through its Office of Research and Development, partially funded and participated in the research described herein. Any opinions expressed in this paper are those of the authors and do not necessarily reflect the views of the agency; therefore, no official endorsement should be inferred. Any mention of trade names or commercial products does not constitute endorsement or recommendation for use.

Author information

Authors and Affiliations

  1. Environmental Protection Agency, Office of Research and Development, Cincinnati, OH, USA

    Aabir Banerji, Jody A. Shoemaker, Dan R. Tettenhorst, Mark Bagley & Jorge W. Santo Domingo

  2. Ramboll Consulting, Arlington, VA, USA

    Ruta Deshpande

  3. Biology Department, University of La Verne, La Verne, CA, USA

    Michael Elk

Authors
  1. Aabir Banerji
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ruta Deshpande
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael Elk
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jody A. Shoemaker
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dan R. Tettenhorst
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mark Bagley
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jorge W. Santo Domingo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jorge W. Santo Domingo.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Consent to publish

The authors hereby attest that this paper is their original unpublished work and is not currently under review elsewhere and hereby consent to its publication within Ecotoxicology.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Banerji, A., Deshpande, R., Elk, M. et al. Highlighting the promise of qPCR-based environmental monitoring: response of the ribosomal RNA:DNA ratio of calanoid copepods to toxic cyanobacteria. Ecotoxicology 30, 411–420 (2021). https://doi.org/10.1007/s10646-021-02366-w

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-021-02366-w

Keywords

Search

Navigation