The FLAMES medium: a new, soft-water culture and bioassay medium for Cladocera

Celis-Salgado, Martha Patricia; Cairns, Allegra; Kim, Natalie; Yan, Norman D.

doi:10.1080/03680770.2008.11902123

Cited by 26 publications

(37 citation statements)

References 21 publications

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“…We cultured Daphnia at 12 °C on a 16L:8D photoperiod in FLAMES zooplankton medium (Celis‐Salgado et al . ) with regular feeding of vitamin‐supplemented Cryptomonas ozolinii . The genotypes were cultured in the laboratory for over one year before the experiments detailed here were conducted.…”

Section: Methodsmentioning

confidence: 99%

Divergence in DNA photorepair efficiency among genotypes from contrasting UV radiation environments in nature

et al. 2015

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Populations of organisms routinely face abiotic selection pressures, and a central goal of evolutionary biology is to understand the mechanistic underpinnings of adaptive phenotypes. Ultraviolet radiation (UVR) is one of earth's most pervasive environmental stressors, potentially damaging DNA in any organism exposed to solar radiation. We explored mechanisms underlying differential survival following UVR exposure in genotypes of the water flea Daphnia melanica derived from natural ponds of differing UVR intensity. The UVR tolerance of a D. melanica genotype from a high-UVR habitat depended on the presence of visible and UV-A light wavelengths necessary for photoenzymatic repair of DNA damage, a repair pathway widely shared across the tree of life. We then measured the acquisition and repair of cyclobutane pyrimidine dimers, the primary form of UVR-caused DNA damage, in D. melanica DNA following experimental UVR exposure. We demonstrate that genotypes from high-UVR habitats repair DNA damage faster than genotypes from low-UVR habitats in the presence of visible and UV-A radiation necessary for photoenzymatic repair, but not in dark treatments. Because differences in repair rate only occurred in the presence of visible and UV-A radiation, we conclude that differing rates of DNA repair, and therefore differential UVR tolerance, are a consequence of variation in photoenzymatic repair efficiency. We then rule out a simple gene expression hypothesis for the molecular basis of differing repair efficiency, as expression of the CPD photolyase gene photorepair did not differ among D. melanica lineages, in both the presence and absence of UVR.

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Section: Methodsmentioning

confidence: 99%

Divergence in DNA photorepair efficiency among genotypes from contrasting UV radiation environments in nature

et al. 2015

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“…First, we compared Bythotrephes survival and reproduction in FLAMES soft-water culture medium, a fully-defined medium for Cladocera that was devised inhouse (Celis-Salgado et al 2008), to water from the source population lake. The major advantage of using an artificial culture medium as opposed to lake water is that the risk of confounding effects arising from biological and chemical variations in culture conditions is minimized.…”

Section: Methods For Rearing the Invasive Zooplankter Bythotrephes Inmentioning

confidence: 99%

“…To determine if we could successfully culture Bythotrephes in a fully defined, softwater medium, a total of 39 F 1 offspring <24 h old were randomly assigned and placed individually into 250 mL of one of the three treatment media: FLAMES medium, 20-µm filtered lake water (FLW), or 80-µm FLW from Peninsula Lake (n = 13 treatment -1 ). FLAMES (CelisSalgado et al 2008) was designed with its chemistry based on two nearby Ontario Ministry of the Environment lakes that support diverse and stable zooplankton assemblages (Yan et al 2008). Lake water was collected every second day in opaque 20-L plastic carboys and filtered in situ through 20-or 80-µm Nitex mesh.…”

Section: Materials and Proceduresmentioning

confidence: 99%

Methods for rearing the invasive zooplankter Bythotrephes in the laboratory

Kim

Yan

2010

Limnology & Ocean Methods

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The invasive spiny water flea's (Bythotrephes longimanus) current North American distribution encompasses the Laurentian Great Lakes as well as a number of inland lakes, particularly on the Canadian Shield. In the past, poor survival in the laboratory has precluded controlled long-term studies on Bythotrephes. Here we investigated field collection techniques and choices of culture media, temperature, and diet that led to the successful maintenance of Bythotrephes from birth to reproduction. Gravid parthenogenic females were collected from invaded lakes. Resulting offspring were reared in source lake water filtered through 20 or 80 µm, or a fully defined artificial culture medium, FLAMES. Individuals raised in FLAMES produced significantly larger broods than those in lake water, indicating that it is an appropriate culture medium. We next conducted a 96-h temperature bioassay on juvenile Bythotrephes. Survival was comparable at 16°C, 20°C, and 24°C but decreased after 48 h at 28°C, and most animals died after 24 h at 32°C. We also reared Bythotrephes at 16°C, 19°C, 22°C, and 25°C. Corresponding intrinsic rates of natural increase (r) for animals maintained to first brood release were 0.02, 0.05, 0.06, and 0.03 d -1 , suggesting that Bythotrephes should be reared at ~22°C to benefit from maximum population increases. Feeding trials confirmed that young Bythotrephes prefer small, slow-moving prey. Finally, we devised a protocol for rearing Bythotrephes that yielded 100% survival to reproduction and r = 0.10 d -1 (for animals maintained to first brood release). Given these results, it is now possible to conduct long-term laboratory studies on this invader.

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“…The animals die below 0.5 mg/L Ca 2+ and their reproductive ability decreases below 1.5 mg/L Ca 2+ (Ashforth and Yan 2008). The 2.5 mg/L Ca 2+ concentration was used to replicate the soft-water FLAMES (Field Laboratory for the Assessment of Multiple Ecological Stressors) medium (Celis-Salgado et al 2008), a medium designed to reflect the chemistry of two Dorset-area, Canadian Shield lakes, Red Chalk and Blue Chalk, which support stable, multispecies assemblages of Daphnia. The highest concentration of 5.0 mg/L Ca 2+ was chosen as a saturation point for native daphniids (Cairns and Yan 2009).…”

Section: Exposure Conditionsmentioning

confidence: 99%

Synergistic interactions of biotic and abiotic environmental stressors on gene expression

Altshuler

McLeod

Colbourne

et al. 2015

Genome

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Understanding the response of organisms to multiple stressors is critical for predicting if populations can adapt to rapid environmental change. Natural and anthropogenic stressors often interact, complicating general predictions. In this study, we examined the interactive and cumulative effects of two common environmental stressors, lowered calcium concentration, an anthropogenic stressor, and predator presence, a natural stressor, on the water flea Daphnia pulex. We analyzed expression changes of five genes involved in calcium homeostasis - cuticle proteins (Cutie, Icp2), calbindin (Calb), and calcium pump and channel (Serca and Ip3R) - using real-time quantitative PCR (RT-qPCR) in a full factorial experiment. We observed strong synergistic interactions between low calcium concentration and predator presence. While the Ip3R gene was not affected by the stressors, the other four genes were affected in their transcriptional levels by the combination of the stressors. Transcriptional patterns of genes that code for cuticle proteins (Cutie and Icp2) and a sarcoplasmic calcium pump (Serca) only responded to the combination of stressors, changing their relative expression levels in a synergistic response, while a calcium-binding protein (Calb) responded to low calcium stress and the combination of both stressors. The expression pattern of these genes (Cutie, Icp2, and Serca) were nonlinear, yet they were dose dependent across the calcium gradient. Multiple stressors can have complex, often unexpected effects on ecosystems. This study demonstrates that the dominant interaction for the set of tested genes appears to be synergism. We argue that gene expression patterns can be used to understand and predict the type of interaction expected when organisms are exposed simultaneously to natural and anthropogenic stressors.

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The FLAMES medium: a new, soft-water culture and bioassay medium for Cladocera

Cited by 26 publications

References 21 publications

Divergence in DNA photorepair efficiency among genotypes from contrasting UV radiation environments in nature

Divergence in DNA photorepair efficiency among genotypes from contrasting UV radiation environments in nature

Methods for rearing the invasive zooplankter Bythotrephes in the laboratory

Synergistic interactions of biotic and abiotic environmental stressors on gene expression

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