A single‐step staining method to evaluate egg viability in zooplankton

Gorokhova, Elena

doi:10.4319/lom.2010.8.414

Cited by 9 publications

(7 citation statements)

References 38 publications

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“…Live females were individually placed in Eppendorf tubes, frozen at −80°C and stored for a few weeks before RNA analysis. For egg viability (EV%; percentage of viable eggs) analysis, all eggs from each well were transferred to a depression slide and stained with TO-PRO-1 iodide (Molecular Probes) [35] . The viable egg production rate (VEPR; viable eggs female −1 day −1 ) was calculated by multiplying EV% with EPR.…”

Section: Methodsmentioning

confidence: 99%

“…To determine EPR, all eggs were counted and related to the number of live females in the corresponding bottle. For EV% measurements, ∼50 eggs per bottle were analyzed with TO-PRO-1 iodide staining [35] . The remaining eggs and nauplii were preserved with acidic Lugol's solution for calculating DI, which incorporates survival and metamorphosis success in copepods: where k i is assigned stage value (in our study: 0 for egg, 1 for nauplii NI, and 2 for nauplii NII; no nauplii had developed beyond NII), n i number of copepods at that stage, and NS – total number of individuals [42] .…”

Section: Methodsmentioning

confidence: 99%

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Bloom-Forming Cyanobacteria Support Copepod Reproduction and Development in the Baltic Sea

et al. 2014

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It is commonly accepted that summer cyanobacterial blooms cannot be efficiently utilized by grazers due to low nutritional quality and production of toxins; however the evidence for such effects in situ is often contradictory. Using field and experimental observations on Baltic copepods and bloom-forming diazotrophic filamentous cyanobacteria, we show that cyanobacteria may in fact support zooplankton production during summer. To highlight this side of zooplankton-cyanobacteria interactions, we conducted: (1) a field survey investigating linkages between cyanobacteria, reproduction and growth indices in the copepod Acartia tonsa; (2) an experiment testing relationships between ingestion of the cyanobacterium Nodularia spumigena (measured by molecular diet analysis) and organismal responses (oxidative balance, reproduction and development) in the copepod A. bifilosa; and (3) an analysis of long term (1999–2009) data testing relationships between cyanobacteria and growth indices in nauplii of the copepods, Acartia spp. and Eurytemora affinis, in a coastal area of the northern Baltic proper. In the field survey, N. spumigena had positive effects on copepod egg production and egg viability, effectively increasing their viable egg production. By contrast, Aphanizomenon sp. showed a negative relationship with egg viability yet no significant effect on the viable egg production. In the experiment, ingestion of N. spumigena mixed with green algae Brachiomonas submarina had significant positive effects on copepod oxidative balance, egg viability and development of early nauplial stages, whereas egg production was negatively affected. Finally, the long term data analysis identified cyanobacteria as a significant positive predictor for the nauplial growth in Acartia spp. and E. affinis. Taken together, these results suggest that bloom forming diazotrophic cyanobacteria contribute to feeding and reproduction of zooplankton during summer and create a favorable growth environment for the copepod nauplii.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Bloom-Forming Cyanobacteria Support Copepod Reproduction and Development in the Baltic Sea

et al. 2014

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“…From this pool, ∼50 eggs bottle −1 were taken for egg viability (% viable eggs) analysis, and the remaining eggs and nauplii were preserved with acid Lugol's solution to estimate egg production and the development index. Egg viability was estimated by staining with TO-PRO-1 iodide (Molecular Probes) which only penetrates eggs with damaged plasma membrane (Gorokhova 2010 ). The development index was calculated according to Knuckey et al ( 2005 ):…”

Section: Methodsmentioning

confidence: 99%

Projected marine climate change: effects on copepod oxidative status and reproduction

Vehmaa

Hogfors

Gorokhova

et al. 2013

Ecology and Evolution

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Zooplankton are an important link between primary producers and fish. Therefore, it is crucial to address their responses when predicting effects of climate change on pelagic ecosystems. For realistic community-level predictions, several biotic and abiotic climate-related variables should be examined in combination. We studied the combined effects of ocean acidification and global warming predicted for year 2100 with toxic cyanobacteria on the calanoid copepod, Acartia bifilosa. Acidification together with higher temperature reduced copepod antioxidant capacity. Higher temperature also decreased egg viability, nauplii development, and oxidative status. Exposure to cyanobacteria and its toxin had a negative effect on egg production but, a positive effect on oxidative status and egg viability, giving no net effects on viable egg production. Additionally, nauplii development was enhanced by the presence of cyanobacteria, which partially alleviated the otherwise negative effects of increased temperature and decreased pH on the copepod recruitment. The interactive effects of temperature, acidification, and cyanobacteria on copepods highlight the importance of testing combined effects of climate-related factors when predicting biological responses.

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“…The fluorophore SYTOX® Green (Invitrogen, Carlsbad, CA) has been used to label dead bacteria (Schumann et al 2003) and eukaryotic algae (Steinberg et al 2011a). Other membranepermeable labels (such as TO-PRO®-1; Invitrogen) have been shown effective for labeling microinvertebrates, specifically, copepod eggs (Gorokhova 2010). Another approach must be used concurrently on the sample to detect the total number of cells.…”

Section: Cell Structural Integritymentioning

confidence: 99%

Comparing a microbial biocide and chlorine as zebra mussel control strategies in an Irish drinking water treatment plant

Meehan¹,

Lucy²,

Gruber³

et al. 2013

MBI

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To minimize the dispersal of aquatic nuisance species through shipping, ballast water can be treated to kill, remove, or inactivate organisms. Ultraviolet (UV) radiation is used in some ballast water management systems to address this goal. Because UV treatment renders cells nonviable (by sterilizing them, preventing reproduction) and does not necessarily kill organisms instantaneously, the efficacy of UV treatment has been verified historically by regrowth assays in which microorganisms are cultured (and thus, reproduce) under optimal growth conditions. Although regrowth assays are definitive, they are time consuming-lasting for days or weeks-and, importantly, are applicable only to organisms amenable to culturing. Furthermore, these estimates of cell concentrations are often accompanied by large error estimates. In this paper, several rapid alternatives to regrowth assays are described and evaluated. An ideal approach would shorten or simplify the analysis burden and, potentially, could be used for shipboard testing to determine compliance with national and international ballast water standards. Complicating this task is the requirement that compliance with the ballast water standards will be determined by quantifying the number of living organisms in ships' ballast water, and while organisms may be living following UV treatment, they may not be viable (i.e., they may not reproduce). To address this dichotomy , alternatives to regrowth assays were categorized based upon the complexity of the analysis and the means used to determine the status of microorganisms (either as viable or living): 1. Instantaneous growth and cell replication, 2. Cell activity and metabolic rates, 3. Cell structural integrity, and 4. Biomolecule presence and status. With the suite of approaches currently available, it is not possible to determine the viability of organisms rapidly, that is, within minutes of collecting a ballast water sample. Measurements of the photosystem integrity via variable fluorescence and the presence of adenosine triphosphate (ATP) are currently the most promising for rapidly estimating concentrations of living cells in compliance testing of ballast water discharges; however, extensive validation is required to verify the applicability of these approaches for the complexity of real-world samples.

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A single‐step staining method to evaluate egg viability in zooplankton

Cited by 9 publications

References 38 publications

Bloom-Forming Cyanobacteria Support Copepod Reproduction and Development in the Baltic Sea

Bloom-Forming Cyanobacteria Support Copepod Reproduction and Development in the Baltic Sea

Projected marine climate change: effects on copepod oxidative status and reproduction

Comparing a microbial biocide and chlorine as zebra mussel control strategies in an Irish drinking water treatment plant

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