69 research outputs found

    The effects of daily cyclic hypoxia on the ecophysiology of the Atlantic ditch shrimp, Palaemon varians

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    Oxygen partial pressure (pO2) is not always constant in aquatic environments and can vary on different timescales, from hours to weeks. In many coastal environments, such as estuaries, lagoons or marshes, pO2 level scan vary on a daily base, resulting in daily cyclic hypoxia. By monitoring temperature and pO2 in the Lymington salt marshes (UK), I was able to quantify diel and seasonal pO2 variability in this coastal habitat:diel oscillations in pO2 were measured in winter, spring and summer, and the greatest pO2 oscillations were recorded in summer, when pO2 could fluctuate from ~42 kPa to ~ 1 kPa every 12 hours, causing diel cyclichypoxia. Even if cyclic hypoxia is common in numerous coastal areas around the world and affects many species, this phenomenon is less studied in comparison to acute or chronic hypoxia. The aim of this thesis was to characterize the short-term effects and the long-term consequences of daily cyclic hypoxia on the physiology of an important decapod crustacean, Palaemon varians. This species was found in the Lymington salt marshes (UK) and, in the laboratory, was subjected to a cyclic hypoxic regime that mimicked conditions measured in the field during summer.In the laboratory, a short 8-hour exposure to hypoxia (pO2 < critical oxygen pressure, pcrit) induced behavioural and metabolic changes and suppressed feeding and ammoniacal excretion. Long-term exposure to diel cyclic hypoxia induced changes in the transcriptome of the animals, prompted an acceleration of the moult cycle (validated at transcriptional and phenotypic level) and eventually resulted in morphological changes to the gills, which increased lamellar surface area. Further, long-term exposure to cyclic hypoxia impaired animal’s growth (in terms of body weight and length), reduced ammoniacal excretion and negatively influenced reproduction by reducing egg yolk content. Interestingly, long-term acclimation to cyclic hypoxia increased thermal tolerance and copper tolerance in comparison to control animals, probably as a consequence of the morphological changes to the gills induced by cyclic hypoxia. Overall, results underline that a short hypoxic exposure repeated daily was able to induce in P. varians alterations at multiple levels of biological organisation. In particular, the observed long-term consequences(i.e. growth reduction, reduced ammoniacal excretion and impaired reproduction) might have important ecological implications for the species and for its ecosystem

    Acclimation to cyclic hypoxia improves thermal tolerance and copper survival in the caridean shrimp Palaemon varians

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    In response to the continuous variation of environmental parameters, species must be able to adjust their physiology to overcome stressful conditions, a process known as acclimatization. Numerous laboratory studies have been conducted to understand and describe the mechanisms of acclimation to one environmental stressor (e.g. cyclic hypoxia), but currently our understanding of how acclimation to one stressor can change tolerance to a subsequent stressor is limited. Here, in two different experiments, we used the shrimp Palaemon varians to test how, following 28-days acclimation to cyclic hypoxia (mimicking a cyclic hypoxic regime currently found in its natural habitat), critical thermal maximum (CT max) and sensitivity to copper (Cu 2+) exposure (30 mgL −1) changed in comparison to shrimp acclimated to normoxic conditions and then exposed to thermal stress or Cu 2+. Acclimation to cyclic hypoxia improved both CT max (~1 °C higher than controls) and survival to acute Cu 2+ exposure (~30% higher than controls) and induced significant gene expression changes (i.e. up-regulation of heat shock protein 70 – HSP70, hypoxia inducible factor – HIF, phosphoenolpyruvate carboxykinase – PEPCK, glucose 6-P transporter – G6Pt, metallothionein – Mt, and down-regulation of hemocyanin – Hem) in animals acclimated to cyclic hypoxia. Our results demonstrate how acclimation to cyclic hypoxia improved tolerance to subsequent stressors, highlighting the complexity of predicting organismal performance in variable (i.e. where multiple parameters can simultaneously change during the day) environments. </p

    Mixtures of environmental pharmaceuticals in marine organisms: Mechanistic evidence of carbamazepine and valsartan effects on Mytilus galloprovincialis

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    Unravelling the adverse outcomes of pharmaceuticals mixture represents a research priority to characterize the risk for marine ecosystems. The present study investigated, for the first time, the interactions between two of the most largely detected pharmaceuticals in marine species: carbamazepine (CBZ) and valsartan (VAL), elucidating mechanisms that can modulate bioaccumulation, excretion and the onset of toxicity. Mytilus galloprovincialis were exposed to environmental levels of CBZ and VAL dosed alone or in combination: measurement of drug bioaccumulation was integrated with changes in the whole transcriptome and responsiveness of various biochemical and cellular biomarkers. Interactive and competing mechanisms between tested drugs were revealed by the much higher CBZ accumulation in mussels exposed to this compound alone, while an opposite trend was observed for VAL. A complex network of responses was observed as variations of gene expression, functional effects on neurotransmission, cell cycle, immune responses and redox homeostasis. The elaboration of results through a quantitative Weight of Evidence model summarized a greater biological reactivity of CBZ compared to VAL and antagonistic interactions between these compounds, resulting in a reduced effect of the antiepileptic when combined with valsartan. Overall, new perspectives are highlighted for a more comprehensive risk assessment of environmental mixtures of pharmaceuticals

    Silencing two main isoforms of crustacean hyperglycemic hormone (CHH) induces compensatory expression of two CHH-like transcripts in the red swamp crayfish Procambarus clarkii

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    RNA interference has frequently been applied to modulate gene function in organisms. With the aim of creating new autocidal methods based on neuro-endocrine disruptors for invasive populations of Procambarus clarkii, we silenced the Crustacean Hyperglycemic Hormone (CHH) by injecting the corresponding dsRNA. CHH is a pleiotropic hormone that primarily regulates the mobilization of energy reserves and plays a pivotal role in stress responses. Here, we describe two experiments aimed at testing whether CHH silencing significantly alters important physiological aspects. The first experiment investigates the effects of CHH silencing at the glycemic and transcriptomic level in the eyestalk. The second experiment explores the long-term effects of CHH silencing and the effects on mortality and moulting rates. Osmotic deficits and mortality were recorded in specimens injected with CHH dsRNA, whilst controls were injected with GFP dsRNA. After 20 days, despite still silenced for CHH, individuals that survived recovered a strong hyperglycemic response after serotonin injection due to the compensatory effect of two peptides belonging to the crustacean neurohormone CHH protein family

    Reproductive plasticity of a Procambarus clarkii population living 10°C below its thermal optimum

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    In this study the annual reproductive biology of a Procambarus clarkii (Girard, 1852) population living in an atypical habitat with cold spring waters is investigated by monitoring Gonado-Somatic and Hepato-Somatic Indexes (GSI and HSI) and by performing cytology on ovaries. Despite its known preference for habitats with water temperature from 21 to 30 °C, our results clearly confirm the adaptation of this population to the atypical thermal habitat, characterised by an annual mean water temperature value of 13.32 ±0.08 °C. Maximum gonadal development was reached in August, with maximum GSI median value of 0.64 (instead of reported values even 10 times higher for other populations), and ovigerous females were found in autumn, with mean realized fecundity of 35 ±7 compared to 285–995 reported from other habitats. Histological analysis was consistent with other studies and allowed us to follow ovarian development at cytological level. The importance of all these results is not to be underestimated: to our knowledge these findings are the first report of the coolest habitat successfully colonized by this species at the present time and so they have to be taken as a warning about the possible range expansion of P. clarkii also to northern and colder habitats that have few things in common with the native habitat of the species and, up to now, were considered “safe” from the invasion of the red swamp crayfish

    Generating gnotobiotic bivalves: a new method on Manila clam (Ruditapes philippinarum)

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    Understanding how microbiomes influence the physiology of animal hosts requires detailed mechanistic insights, often obtained through gnotobiological approaches. Model organisms are central to this research, offering key advantages such as experimental tractability, reproducibility, and ease of manipulation. However, there is a lack of established gnotobiotic models for the marine environment-especially for bivalves, which play a critical role in ecosystem functioning. This gap is particularly important in the context of climate change, where harnessing microbiome resilience could mitigate environmental challenges and enhance host responses. In this study, we present a protocol to generate microbiome-depleted and gnotobiotic clams (Ruditapes philippinarum), one of the most widely farmed molluscs in the world and a key sentinel species for environmental and climate change impacts. Our microbiome depletion protocol effectively eliminated all detectable bacterial genera in the clams, with the exception of Endozoicomonas elysicola, which was identified solely by 16S rRNA amplicon sequencing and not by cultivation methods. In addition, we developed a microbiome transplantation protocol using inoculation of a mock bacterial community that successfully colonized the recipient clams within 1 h of transplantation. By extending gnotobiotic methods to marine invertebrates, this work opens new avenues for investigating microbial influences on ecologically and economically important species, particularly under the pressure of a changing climate.IMPORTANCEThe extensive diversity of host-microbe symbioses across ecosystems requires the use of different models to identify conserved and specific processes underlying such relationships. The need for novel models is particularly relevant in the context of the rapid environmental modifications due to climate change. Bivalve molluscs play a crucial role in the functioning of marine ecosystems. In this study, we present the first experimental protocol for the generation of gnotobiotic clams of the species Ruditapes philippinarum, one of the most widely farmed molluscs in the world, and a sentinel organism for environmental pollution. Our work extends the current technical understanding of the establishment of gnotobiotic animals, providing an important method for testing research hypotheses on a key taxonomic group in animal ecology. This study will also open new avenues for investigating the influence of microorganisms on animal health and elucidate the transferability of mechanisms studied predominantly in vertebrates to marine invertebrates

    The consequences of daily cyclic hypoxia on a European grass shrimp: from short-term responses to long-term effects

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    1.Salt marshes are a key coastal environment for their important role as nursery habitats for marine and estuarine fish and crustaceans. Salt marshes are variable environments where species can experience daily cyclic hypoxic stress, characterized by profound variations in oxygen partial pressure (pO2) from supersaturated conditions (~42kPa) to extremely hypoxic conditions (~3kPa) in ~12‐hours. 2.Here, under laboratory conditions, we assessed the physiological consequences of exposing the shrimp Palaemon varians, a species commonly found in the salt marshes of northern Europe, to the daily cyclic hypoxic regime currently experienced in its habitat in August (7.1±1.8 hours day−1 below 4.0kPa). In the laboratory adults were kept at water pO2 &lt;4.5kPa for 7‐hours each night and in normoxic conditions for the rest of the time. 3.We recorded an acceleration of P. varians’ moult cycle, which was 15% shorter in animals kept in cyclic hypoxia compared to animals in normoxia. Similarly, the pattern of expression of two cuticular proteins over an entire moult cycle indicated an effect of cyclic hypoxia on moult stage‐related genes. After 16 days, morphological changes to the gills were detected, with shrimps in cyclic hypoxia having a 13.6% larger lamellar surface area (measured in μm2/mg animal) than normoxic animals, which could improve gas exchange capacity. Overall, phenotypic and morphological data indicate that faster moulting is triggered in response to cyclic hypoxia, with the benefit that gill modifications can be prompted more rapidly in order to meet oxygen requirements of the body. 4.On the first experimental day, in cyclic hypoxic exposed animals, we recorded a 50% decrease in feeding rates (during hypoxic conditions) in comparison to normoxic animals. Similarly, ammonium excretion was reduced by 66‐75% during the 1st and 21st experimental day. Body size was reduced by ~4% after 28 days. Females that reproduced in cyclic hypoxic conditions reduced the amount of yolk in each egg by ~24%. Overall, results underline how, in a decapod shrimp living in a key coastal environment, many physiological parameters are impaired by a cyclic hypoxic regime that is currently found in its natural habitat

    Reproductive plasticity of a Procambarus clarkii population living 10°C below its thermal optimum

    No full text
    In this study the annual reproductive biology of a Procambarus clarkii (Girard, 1852) population living in an atypical habitat with cold spring waters is investigated by monitoring Gonado-Somatic and Hepato-Somatic Indexes (GSI and HSI) and by performing cytology on ovaries. Despite its known preference for habitats with water temperature from 21 to 30 °C, our results clearly confirm the adaptation of this population to the atypical thermal habitat, characterised by an annual mean water temperature value of 13.32 ±0.08 °C. Maximum gonadal development was reached in August, with maximum GSI median value of 0.64 (instead of reported values even 10 times higher for otherpopulations), and ovigerous females were found in autumn, with mean realized fecundity of 35 ±7 compared to 285–995 reported from other habitats. Histological analysis was consistent with other studies and allowed us to follow ovarian development at cytological level. The importance of all these results is not to be underestimated: to our knowledge these findings are the first report of the coolest habitat successfully colonized by this species at the present time and so they have to be taken as a warning about the possible range expansion of P.clarkii also to northern and colder habitats that have few things in common with the native habitat of the species and, up to now, were considered “safe” from the invasion of the red swamp crayfish

    Data from: The consequences of daily cyclic hypoxia on a European grass shrimp: from short-term responses to long-term effects

    No full text
    1. Salt marshes are a key coastal environment for their important role as nursery habitats for marine and estuarine fish and crustaceans. Salt marshes are variable environments where species can experience daily cyclic hypoxic stress, characterized by profound variations in oxygen partial pressure (pO2) from supersaturated conditions (~42kPa) to extremely hypoxic conditions (~3kPa) in ~12-hours. 2. Here, under laboratory conditions, we assessed the physiological consequences of exposing the shrimp Palaemon varians, a species commonly found in the salt marshes of northern Europe, to the daily cyclic hypoxic regime currently experienced in its habitat in August (7.1&plusmn;1.8 hours day-1 below 4.0kPa). In the laboratory adults were kept at water pO2 &lt;4.5kPa for 7-hours each night and in normoxic conditions for the rest of the time. 3. We recorded an acceleration of P. varians&rsquo; moult cycle, which was 15% shorter in animals kept in cyclic hypoxia compared to animals in normoxia. Similarly, the pattern of expression of two cuticular proteins over an entire moult cycle indicated an effect of cyclic hypoxia on moult stage-related genes. After 16 days, morphological changes to the gills were detected, with shrimps in cyclic hypoxia having a 13.6% larger lamellar surface area (measured in &micro;m2/mg animal) than normoxic animals, which could improve gas exchange capacity. Overall, phenotypic and morphological data indicate that faster moulting is triggered in response to cyclic hypoxia, with the benefit that gill modifications can be prompted more rapidly in order to meet oxygen requirements of the body. 4. On the first experimental day, in cyclic hypoxic exposed animals, we recorded a 50% decrease in feeding rates (during hypoxic conditions) in comparison to normoxic animals. Similarly, ammonium excretion was reduced by 66-75% during the 1st and 21st experimental day. Body size was reduced by ~4% after 28 days. Females that reproduced in cyclic hypoxic conditions reduced the amount of yolk in each egg by ~24%. Overall, results underline how, in a decapod shrimp living in a key coastal environment, many physiological parameters are impaired by a cyclic hypoxic regime that is currently found in its natural habitat. </span
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