Symbiotic microalgae do not increase susceptibility of zooxanthellate medusae (Cassiopea xamachana) to herbicides.

Herbicides are among the most detected pesticides in coastal environments. Herbicides may impact non-target organisms, but invertebrates that have a symbiotic relationship with microalgae (zooxanthellae) may be particularly susceptible. How zooxanthellae influence the response of organisms to herbicides, however, remains untested. We exposed zooxanthellate and azooxanthellate Cassiopea xamachana medusae to environmentally relevant concentrations of the herbicide atrazine (0 µg L - 1, 7 µg L - 1 and 27 µg L - 1) for 20 days. We hypothesised that atrazine would have adverse effects on the size, rate of bell contractions and, respiration of medusae, but that effects would be more severe in zooxanthellate than azooxanthellate medusae. We also predicted that photosynthetic efficiency, chlorophyll a (Chla) content and zooxanthellae density would decrease in zooxanthellate medusae exposed to atrazine. Both zooxanthellate and azooxanthellate medusae shrank, yet the size-specific respiration rates were not constant during the experiment. Photosynthetic efficiency of zooxanthellate medusae significantly decreased at 7 and 27 µgL-1 atrazine, but atrazine did not affect the Chla content or zooxanthellae density. Our results showed that even though atrazine inhibited photosynthesis, zooxanthellae were not expelled from the host. We conclude that the presence of zooxanthellae did not increase the susceptibility of C. xamachana medusae to atrazine.

Chemicals sorbed to environmental microplastics are toxic to early life stages of aquatic organisms.

Microplastics are ubiquitous in aquatic ecosystems, but little information is currently available on the dangers and risks to living organisms. In order to assess the ecotoxicity of environmental microplastics (MPs), samples were collected from the beaches of two islands in the Guadeloupe archipelago, Petit-Bourg (PB) located on the main island of Guadeloupe and Marie-Galante (MG) on the second island of the archipelago. These samples have a similar polymer composition with mainly polyethylene (PE) and polypropylene (PP). However, these two samples are very dissimilar with regard to their contamination profile and their toxicity. MPs from MG contain more lead, cadmium and organochlorine compounds while those from PB have higher levels of copper, zinc and hydrocarbons. The leachates of these two samples of MPs induced sublethal effects on the growth of sea urchins and on the pulsation frequency of jellyfish ephyrae but not on the development of zebrafish embryos. The toxic effects are much more marked for samples from the PB site than those from the MG site. This work demonstrates that MPs can contain high levels of potentially bioavailable toxic substances that may represent a significant ecotoxicological risk, particularly for the early life stages of aquatic animals.

Modulatory Activities of Plant Extracts on Jellyfish Cytotoxicity.

INTRODUCTION: The potential efficacy of selected plant extracts to counteract the dermal toxicity of jellyfish envenomation was investigated using an in vitro cell culture model. METHODS: We studied plant extracts from Carica papaya, Ananas comosus, and Bouvardia ternifolia, known for their antivenom properties, in pairwise combinations with tissue homogenates of the jellyfish Pelagia noctiluca, Phyllorhiza punctata, and Cassiopea andromeda, to evaluate modulations of jellyfish cytotoxic effects. L929 mouse fibroblasts were incubated with pairwise jellyfish/plant extract combinations and examined by MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide). RESULTS: C papaya and A comosus significantly lowered the cytotoxicity of P noctiluca and P punctata but induced a slight worsening of C andromeda cytotoxicity. Conversely, B ternifolia was protective against P punctata, ineffective against P noctiluca, and worsened C andromeda cytotoxicity. CONCLUSIONS: Data showed species-specific and contrasting effects of plant extracts, suggesting that those containing protease activities, namely A comosus and C papaya, are more effective in lowering the cytotoxicity of jellyfish venom containing toxic peptidic factors such as phospholipase A. However, all examined plants require further investigation in vivo to evaluate their ability to counteract jellyfish injury to the skin.

Polyps of the Jellyfish Aurelia aurita Are Unaffected by Chronic Exposure to a Combination of Pesticides.

Pesticides are a major contaminant in coastal waters and can cause adverse effects in marine invertebrates such as jellyfish. Most studies have investigated short-term responses of organisms to unrealistically high concentrations of pesticides; however, chronic exposure to persistent low concentrations, which are more likely to occur in the environment, are rarely analyzed. We tested the response of polyps of the moon jellyfish Aurelia aurita to environmental concentrations of the herbicide atrazine and the insecticide chlorpyrifos, individually and in combination, over 9 wk. We hypothesized that exposure to individual pesticides would reduce rates of asexual reproduction and alter polyps' metabolite profiles, and that the results would be more severe when polyps were exposed to the combined pesticides. Polyps survived and reproduced (through budding) in all treatments, and no differences among treatments were observed. Proton nuclear magnetic resonance spectroscopy revealed no difference in profiles of polar metabolites of polyps exposed to the individual or combined pesticides. Our results suggest that A. aurita polyps are unaffected by chronic exposure to atrazine and chlorpyrifos at concentrations recommended as being protective by current Australian water quality guidelines. Environ Toxicol Chem 2020;39:1685-1692. © 2020 SETAC.

Detecting effects of herbicide runoff: The use of Cassiopea maremetens as a biomonitor to hexazinone.

Herbicides are an integral part of global agricultural activity but can be advected into local drainages that can discharge to coastal marine systems. Herbicide runoff can impact coastal marine organisms, including those associated with coral reefs and coastal mangrove forests. In this study, the symbiotic sedentary jellyfish Cassiopea maremetens were exposed to analytical grade hexazinone to determine their sensitivity and potential for recovery after exposure to a press herbicide event of 14 days followed by a recovery period of matching duration. Bell surface area, photosynthetic yield (i.e. effective quantum yield, EQY), statolith count and zooxanthellae density were analyzed. Most metrics demonstrated significant decreases when exposed to higher concentrations of hexazinone, while EQY was significantly decreased at exposure concentrations from 31 µg/L hexazinone and above. In contrast, zooxanthellae density (cells/mm2) increased in the highest concentrations compared to control animals. At the end of the exposure period the EC50 for bell surface area, EQY, and statolith count were 176 µg/L, 81.96 µg/L, and 304.3 µg/L, respectively. Jellyfish were able to recover to similar start values for all measured metrics at the end of the 14-day recovery period, with EQY showing recovery by Day 7 of the recovery period. This study demonstrated that statolith counts as an estimate of age were not affected by herbicides. We conclude that the depressed metrics from herbicide related impacts of C. maremetens are effective indicators of a relatively recent herbicide perturbation in that the recovery timeframe for these jellyfish is relatively short.

Microplastics ingestion in the ephyra stage of Aurelia sp. triggers acute and behavioral responses.

For the first time, we report a correspondence between microplastics (MP) ingestion and ecotoxicological effects in gelatinous zooplankton (Cnidarian jellyfish). The ephyra stage of the jellyfish Aurelia sp. was exposed to both environmental and high concentrations of fluorescent 1-4 µm polyethylene MP (0.01-10 mg/L). After 24 and 48 h, MP accumulation, acute (Immobility) and behavioral (Frequency pulsation) endpoints were investigated. MP were detected by confocal and tomographic investigations on gelatinous body and mouth, either attached on the surface or ingested. This interaction was responsible for impairing ephyrae survival and behavior at all tested concentrations after 24 h. Acute and behavioral effects were also related to mechanical disturbance, caused by MP, triggering a loss of radial symmetry. Contaminated ephyrae exposed to clean seawater showed full recovery after 72 h highlighting the organisms without the microspheres, attached on body jellyfish surface around the mouth and lappets. In conclusion, short-term exposure to MP affects ephyrae jellyfish health, impairing both their survival and behavior. Polyethylene MP temporarily affect both Immobility and Frequency of pulsation of Aurelia sp. jellyfish. This study provides a first step towards understanding and clarifying the potential impacts of MP contamination in gelatinous zooplankton.

EVALUATION OF EUTHANASIA OF MOON JELLYFISH (AURELIA AURITA) USING SIMPLE SALT SOLUTIONS.

Immersion euthanasia methods reported over the most recent decades for aquatic invertebrates use organic alcohols or halogenated hydrocarbons that can interfere with nuclear magnetic resonance (NMR) analysis. A rolling study design evaluated potassium chloride (KCl), magnesium chloride (MgCl2), and magnesium sulfate (MgSO4) as potential ion-based euthanasia methods for moon jellyfish (Aurelia aurita) destined for metabolomic analysis by NMR spectroscopy. Death was defined as the cessation of autonomous bell pulsing and response to external stimulus. MgCl2 applied at a dose of 142 g/L provided euthanasia within 32 sec of applications without the untoward effects observed with the other two salts. Euthanasia with KCl at the doses tested was associated with abnormal behavior and tissue degradation during dissection. MgSO4 at the doses tested resulted in abnormal behavior and failed to provide rapid euthanasia.

An investigation on the impact of substrate type, temperature, and iodine on moon jellyfish production.

Jellyfish are a popular public aquarium species, however, their collection from natural populations is undesirable due to impact on species abundance and bycatch. Thus, a sustainable supply of jellyfish bred in-house would be highly desirable. Here we describe an investigation into developing a sustainable moon jellyfish, Aurelia aurita, breeding program by determining the impact of substrate type on reattachment of polyps and the influence of iodine and temperature on strobilation and ephyra production. To test whether reattachment and growth of moon jellyfish polyps are influenced by substrate type polyps were provided with anthropogenic and natural substrates after being dislodged in experimental aquaria. Polyps selectively re-attached to plastics rather than natural materials. However, polyp growth was similar on all tested substrates. We tested whether cooling and addition of iodine affected strobilation. A period of cooling of around 10 °C while also introducing soluble iodine to the polyps enhanced strobilation rate. This method produced ephyra at a reliable rate in captivity negating the need for collection of wild individuals providing a supply of individuals for exhibit and for conservation research within a public aquarium. These results demonstrate that plastics should be adopted as an easier to colonize substrate and the use of cooling with iodine addition can enhance sustainable breeding protocols of moon jellyfish and may be relevant to the production of comparable jellyfish species.

Metabolic and oxidative stress responses of the jellyfish Cassiopea to pollution in the Gulf of Aqaba, Jordan.

Physiological responses of jellyfish to pollution are virtually overlooked. We measured the activity of two glycolytic enzymes (pyruvate kinase (PK) and lactate dehydrogenase (LDH)), lipid peroxidation (LPO), protein and chlorophyll a content in the jellyfish Cassiopea sp. from polluted and reference sites along the Gulf of Aqaba, Jordan. In jellyfish from polluted sites, low PK/LDH ratios and high LDH activity clarify their reliance on anaerobic metabolism. PK and LDH were positively correlated in the jellyfish. While medusae from polluted sites showed no signs of oxidative stress damage, protein content was significantly lower. This might suggest protein utilization for energy production needed for maintenance. Unchanged LPO in polluted sites indicates the ability of jellyfish to keep reactive oxygen species under control. Overall these results suggest that the jellyfish seems to tolerate the current levels of pollution at the studied sites and they might be anaerobically poised to live at such habitats.

Effect of tea saponin on ephyrae and polyps of the moon jellyfish Aurelia sp.1.

The moon jellyfish (Aurelia sp.1) is thought to be a nuisance for the sea cucumber aquaculture, which commonly occur in the sea cucumber (Apostichopus japonicus) culture ponds of the Yellow Sea, China. To develop an appropriate method to control Aurelia sp.1 blooms, the toxic effects of tea saponin on Aurelia sp.1 ephyrae and polyps were tested in laboratory experiments. Our results revealed that tea saponin caused significant morphological changes, behavioral abnormality and mortality in Aurelia sp.1 ephyrae and polyps in 24 h and 48 h exposure experiments. The 24 h and 48 h median lethal concentrations (LC50) values of tea saponin for Aurelia sp.1 ephyrae were 1.9 and 1.1 mg L-1 respectively, while the LC50 value for Aurelia sp.1 polyps was 0.4 mg L-1 after 24h and 48 h of exposure to tea saponin. Comparison with literature results of tea saponin on A. japonicus indicates that the resistance of A. japonicus to tea saponin is 12-18 times greater than that of Aurelia sp.1 ephyrae. Therefore, the appropriate tea saponin dosage for the control of Aurelia sp.1 should be paid enough attention in order to minimize possible damage for sea cucumber. We suggest that the recommended level of tea saponin to eradicate Aurelia sp.1 ephyrae and polyps in sea cucumber culture ponds be lower than 1.35 mg L-1.

Herbicide effects on the growth and photosynthetic efficiency of Cassiopea maremetens.

Herbicides from agricultural run-off have been measured in coastal systems of the Great Barrier Reef over many years. Non-target herbicide exposure, especially photosystem II herbicides has the potential to affect seagrasses and other marine species. The symbiotic benthic jellyfish Cassiopea maremetens is present in tropical/sub-tropical estuarine and marine environments. Jellyfish (n = 8 per treatment) were exposed to four separate concentrations of agricultural formulations of diuron or hexazinone to determine their sensitivity and potential for recovery to pulsed herbicide exposure. Jellyfish growth, symbiont photosynthetic activity and zooxanthellae density were analysed for herbicide-induced changes for 7 days followed by a 7 day recovery period. Both the jellyfish and endosymbiont were more sensitive to diuron than hexazinone. The 7-day EC50 for jellyfish growth was 0.35 µg L-1 for Diuron and 17.5 µg L-1 for Hexazinone respectively. Diuron exposure caused a significant decrease (p < 0.05) in jellyfish growth at 0.1 µg L-1, a level that is below the regional Great Barrier Reef guideline value. Jellyfish recovery was rapid with growth rates similar to control animals following removal from herbicide exposure. Both diuron and hexazinone caused significant decreases in photosynthetic efficiency (effective quantum yield) in all treatment concentrations (0.1 µg L-1 and above) and this effect continued in the post-exposure period. As this species is frequently found in near-shore environments, they may be particularly vulnerable to herbicide run-off.

To Pee, or Not to Pee: A Review on Envenomation and Treatment in European Jellyfish Species.

There is a growing cause for concern on envenoming European species because of jellyfish blooms, climate change and globalization displacing species. Treatment of envenomation involves the prevention of further nematocyst release and relieving local and systemic symptoms. Many anecdotal treatments are available but species-specific first aid response is essential for effective treatment. However, species identification is difficult in most cases. There is evidence that oral analgesics, seawater, baking soda slurry and 42-45 °C hot water are effective against nematocyst inhibition and giving pain relief. The application of topical vinegar for 30 s is effective on stings of specific species. Treatments, which produce osmotic or pressure changes can exacerbate the initial sting and aggravate symptoms, common among many anecdotal treatments. Most available therapies are based on weak evidence and thus it is strongly recommended that randomized clinical trials are undertaken. We recommend a vital increase in directed research on the effect of environmental factors on envenoming mechanisms and to establish a species-specific treatment. Adequate signage on jellyfish stings and standardized first aid protocols with emphasis on protective equipment and avoidance of jellyfish to minimize cases should be implemented in areas at risk.

The use of ephyrae of a scyphozoan jellyfish, Aurelia aurita, in the aquatic toxicological assessment of Macondo oils from the Deepwater Horizon incident.

Ephyrae of the scyphozoan jellyfish, Aurelia aurita, were evaluated in 96-hr acute toxicity tests for lethal response to Macondo crude oils from the Deepwater Horizon (DWH) incident in the Gulf of Mexico (GOM), Corexit 9500, and oil-dispersant mixtures. Water accommodated fractions (WAFs) of weathered and unweathered Macondo crude oils were not acutely toxic to ephyrae (LC50s > 100% WAF). The total PAHs (TPAHs), measured as the sum of 46 PAHs, averaged 21.1and 152 µg TPAH/L for WAFs of weathered and unweathered oil, respectively. Mortality was significantly (p = <0.0001) higher in the three highest exposure concentrations (184-736 µg TPAH/L) of chemically dispersed WAFs (CEWAF) compared to controls. Dispersant only tests resulted in a mean LC50 of 32.3 µL/L, which is in the range of previously published LC50s for marine zooplankton. Changes in appearance and muscle contractions were observed in organisms exposed to CEWAF dilutions of 12.5 and 25%, as early as 24 h post-exposure. Based on the results of these tests, crude oil alone did not cause significant acute toxicity; however, the presence of chemical dispersant resulted in substantial mortality and physical and behavioral abnormalities either due to an increase in hydrocarbons or droplet exposure.

Reduced salinity increases susceptibility of zooxanthellate jellyfish to herbicide toxicity during a simulated rainfall event.

Accurately predicting how marine biota are likely to respond to changing ocean conditions requires accurate simulation of interacting stressors, exposure regimes and recovery periods. Jellyfish populations have increased in some parts of the world and, despite few direct empirical tests, are hypothesised to be increasing because they are robust to a range of environmental stressors. Here, we investigated the effects of contaminated runoff on a zooxanthellate jellyfish by exposing juvenile Cassiopea sp. medusae to a photosystem II (PSII) herbicide, atrazine and reduced salinity conditions that occur following rainfall. Four levels of atrazine (0ngL(-1), 10ngL(-1), 2µgL(-1), 20µgL(-1)) and three levels of salinity (35 ppt, 25 ppt, 17 ppt) were varied, mimicking the timeline of light, moderate and heavy rainfall events. Normal conditions were then slowly re-established over four days to mimic the recovery of the ecosystem post-rain and the experiment continued for a further 7 days to observe potential recovery of the medusae. Pulse-amplitude modulated (PAM) chlorophyll fluorescence, growth and bell contraction rates of medusae were measured. Medusae exposed to the combination of high atrazine and lowest salinity died. After 3 days of exposure, bell contraction rates were reduced by 88% and medusae were 16% smaller in the lowest salinity treatments. By Day 5 of the experiment, all medusae that survived the initial pulse event began to recover quickly. Although atrazine decreased YII under normal salinity conditions, YII was further reduced when medusae were exposed to both low salinity and atrazine simultaneously. Atrazine breakdown products were more concentrated in jellyfish tissues than atrazine at the end of the experiment, suggesting that although bioaccumulation occurred, atrazine was metabolised. Our results suggest that reduced salinity may increase the susceptibility of medusae to herbicide exposure during heavy rainfall events.

Jellyfish model for ototoxicity.

OBJECTIVE: Pharmacologic ototoxicity is well described in the medical literature, yet efficient screening models are lacking. Aurelia aurita ephyrae, transparent jellyfish with identifiable hair cells, could be an effective model. Structural changes readily manifest behaviorally, and hair cells are easily stained and observed. We treated ephyrae with various gentamicin concentrations, evaluated its motility, and quantified its hair cell loss. STUDY DESIGN: Baseline pulsing per minute (P), swimming (S), and orientation (O) values were recorded from cultured ephyrae. Ephyrae were transferred into test tubes containing artificial seawater (ASW), gentamicin, or penicillin. P, S, and O were scored at 0, 24, and 48 hours. Ephyrae were formalin fixed, phalloidin stained, and imaged with confocal microscopy, and hair cells were then counted. RESULTS: P was impaired by gentamicin in a dose-dependent fashion, whereas ASW controls maintained baseline P, S, and O values. Impairment of S and O occurred with 3.5 mmol/L gentamicin at 24 hours. For six experiments each using 40 ephyrae, at 24 hours, average P was reduced from 75.2 in ASW to 28.8, 12.3, and 1.9 for 1, 2, and 3.5 mmol/L gentamicin, respectively (p < 0.05 for all cases). Hair cell loss at 24 and 48 hours was significant (32% and 48% reduction compared with control, p < 0.05) and correlated with motility deficits. Deficits from penicillin exposure were not statistically significant. CONCLUSION: The ephyra model demonstrated functional and histologic gentamicin-mediated impairments, showing promise as a screening tool for ototoxic agents. The changes in ephyra motility after gentamicin exposure correlated significantly with hair cell loss.

Ephyra jellyfish as a new model for ecotoxicological bioassays.

The aim of this study was a preliminary investigation on the possibility of using the ephyra of Scyphozoan jellyfish Aurelia aurita (Linnaeus, 1758), the common moon jellyfish, as an innovative model organism in marine ecotoxicology. A series of sequential experiments have been carried out in laboratory in order to investigate the influence of different culturing and methodological parameters (temperature, photoperiod, ephyrae density and age) on behavioural end-points (% of Frequency of Pulsations) and standardize a testing protocol. After that, the organisms have been exposed to two well known reference toxic compounds (Cadmium Nitrate and SDS) in order to analyse the acute and behavioural responses during static exposure. Results of this work indicate that the proposed behavioural end-point, frequency of pulsations (Fp), is an easily measurable one and can be used coupled with an acute one (immobilization) and that ephyrae of jellyfish are very promising model organisms for ecotoxicological investigation.

Sea water acidification affects osmotic swelling, regulatory volume decrease and discharge in nematocytes of the jellyfish Pelagia noctiluca.

BACKGROUND: Increased acidification/PCO2 of sea water is a threat to the environment and affects the homeostasis of marine animals. In this study, the effect of sea water pH changes on the osmotic phase (OP), regulatory volume decrease (RVD) and discharge of the jellyfish Pelagia noctiluca (Cnidaria, Scyphozoa) nematocytes, collected from the Strait of Messina (Italy), was assessed. METHODS: Isolated nematocytes, suspended in artificial sea water (ASW) with pH 7.65, 6.5 and 4.5, were exposed to hyposmotic ASW of the same pH values and their osmotic response and RVD measured optically in a special flow through chamber. Nematocyte discharge was analyzed in situ in ASW at all three pH values. RESULTS: At normal pH (7.65), nematocytes subjected to hyposmotic shock first expanded osmotically and then regulated their cell volume within 15 min. Exposure to hyposmotic ASW pH 6.5 and 4.5 compromised the OP and reduced or totally abrogated the ensuing RVD, respectively. Acidic pH also significantly reduced the nematocyte discharge response. CONCLUSION: Data indicate that the homeostasis and function of Cnidarians may be altered by environmental changes such as sea water acidification, thereby validating their use as novel bioindicators for the quality of the marine environment.

Indomethacin induction of metamorphosis from the asexual stage to sexual stage in the moon jellyfish, Aurelia aurita.

We found while screening a chemical library that indomethacin, an inhibitor of prostaglandin biosynthesis, induced strobilation (metamorphosis from the asexual to sexual stage) in the moon jellyfish, Aurelia aurita. Indomethacin initiated strobilation in a dose-dependent manner, but was not involved in the progression of strobilation. Pharmacological experiments suggested that indomethacin could induce strobilation independently of prostaglandin biosynthesis.

Nematocytes' activation in Pelagia noctiluca (Cnidaria, Scyphozoa) oral arms.

Nematocytes' discharge is triggered to perform both defense and predation strategies in cnidarians and occurs under chemico-physical stimulation. In this study, different compounds such as amino acids and proteins (mucin, albumin, poly-L: -lysine, trypsin), sugars and N-acetylate sugars (N-acetyl neuraminic acid, N-acetyl galactosamine, sucrose, glucose, agarose and trehalose), nucleotides (ATP and cAMP), were tested as chemosensitizers of nematocyte discharge in the oral arms of the scyphozoan Pelagia noctiluca, particularly abundant in the Strait of Messina (Italy). Excised oral arms were submitted to a combined chemico-physical stimulation by treatment with different compounds followed by mechanical stimulation by a non-vibrating test probe. Discharge induced by a chemico-physical stimulation was more significant than that obtained after mechanical stimulation alone. A chemosensitizing mechanism, with a dose-dependent effect, was observed after treatment with sugars, amino compounds such as glutathione, nucleotides and mucin, according to that already seen in sea anemones. Such findings suggest that, though Anthozoa and Scyphozoa exhibit different divergence times during the evolutionary process, the discharge activation exhibits common features, probably derived from their last common ancestor.

Multicellular redox regulation in an early-evolving animal treated with glutathione.

Redox signaling has emerged as a unifying theme in many seemingly disparate disciplines. Such signaling has been widely studied in bacteria and eukaryotic organelles and is often mediated by reactive oxygen species (ROS). In this context, reduced glutathione (GSH) acts as an important intracellular antioxidant, diminishing ROS and potentially affecting redox signaling. Complementing this cell-level perspective, colonial hydroids can be a useful model for understanding organism-level redox signaling. These simple, early-evolving animals consist of feeding polyps connected by tubelike stolons. Colonies treated exogenously with GSH or reduced glutathione ethyl ester (GEE) were expected to show a morphological change to sheetlike growth typical of low levels of ROS. Contrary to expectations, diminished stolon branching and polyp initiation was observed. Such runnerlike growth is associated with higher levels of ROS, and surprisingly, such higher levels were found in GSH- and GEE-treated colonies. Further investigations show that GSH triggered a feeding response in hydroid polyps, increasing oxygen uptake but at the same time relaxing mitochondrion-rich contractile regions at the base of polyps. Diminished gastrovascular flow and increased emissions of mitochondrial ROS also correlated with the observed runnerlike growth. In contrast to cell-level, "bottom-up" views of redox signaling, here the phenotype may arise from a "top-down" interaction of mitochondrion-rich regions and organism-level physiology. Such multicellular redox regulation may commonly occur in other animals as well.