Occurrence of the pugnose pipefish Bryx dunckeri in the Sargasso Sea.

Juvenile specimens of the pugnose pipefish, Bryx dunckeri, were collected during a multipurpose research survey conducted within the Sargasso Sea Subtropical Convergence Zone, extending the known distribution range of this species to include open ocean areas of the Western North Atlantic. Novel spatial data are of scientific interest as information on the distribution, population structure, and population size of this species is limited. Additionally, we present detailed photographs and morphological data on the collected specimens. The results are discussed in relation to the dispersal abilities and population structure in syngnathids.

Open science.

The term 'open science' refers to a range of methods, tools, platforms and practices that aim to make scientific research more accessible, transparent, reproducible and reliable. This includes, for example, sharing code, data and research materials, embracing new publishing formats such as registered reports and preprints, pursuing replication studies and reanalyses, optimising statistical approaches to improve evidence assessment and re-evaluating institutional incentives. The ongoing shift towards open science practices is partly due to mounting evidence that studies across disciplines suffer from biases, underpowered designs and irreproducible or non-replicable results. It also stems from a general desire amongst many researchers to reduce hyper-competitivity in science and instead promote collaborative research that benefits science and society.

A novel method for measuring acute thermal tolerance in fish embryos.

Aquatic ectotherms are vulnerable to thermal stress, with embryos predicted to be more sensitive than juveniles and adults. When examining the vulnerability of species and life stages to warming, comparable methodology must be used to obtain robust conclusions. Critical thermal methodology is commonly used to characterize acute thermal tolerances in fishes, with critical thermal maximum (CTmax) referring to the acute upper thermal tolerance limit. At this temperature, fish exhibit loss of controlled locomotion due to a temperature-induced collapse of vital physiological functions. While it is relatively easy to monitor behavioural responses and measure CTmax in larval and adult fish, this is more challenging in embryos, leading to a lack of data on this life stage, or that studies rely on potentially incomparable metrics. Here, we present a novel method for measuring CTmax in fish embryos, defined by the temperature at which embryos stop moving. Additionally, we compare this measurement with the temperature of the embryos' last heartbeat, which has previously been proposed as a method for measuring embryonic CTmax. We found that, like other life stages, late-stage embryos exhibited a period of increased activity, peaking approximately 2-3°C before CTmax. Measurements of CTmax based on last movement are more conservative and easier to record in later developmental stages than measurements based on last heartbeat, and they also work well with large and small embryos. Importantly, CTmax measurements based on last movement in embryos are similar to measurements from larvae and adults based on loss of locomotory control. Using last heartbeat as CTmax in embryos likely overestimates acute thermal tolerance, as the heart is still beating when loss of response/equilibrium is reached in larvae/adults. The last movement technique described here allows for comparisons of acute thermal tolerance of embryos between species and across life stages, and as a response variable to treatments.

Effects of the invasive swim bladder parasite Anguillicola crassus on health and condition indicators in the European eel.

Parasites negatively affect biological processes within their hosts, which may alter for example health, growth, and reproductive ability. Non-native invasive parasites, in particular, may have large effects on the endemic hosts, given that the hosts lack evolved specific defences against such parasites. The swim bladder nematode Anguillicola crassus, an invasive parasite originating from Asia, is found in the European eel (Anguilla anguilla, L. 1758), since the 1980s. We investigated whether A. crassus affected several indicators related to health of the European eel (spleen- and liver size, body fat content and relative condition). Our results indicate that during the continental residency of the eels, infection by A. crassus had no major negative impacts on the investigated health indicators at the generally low infection intensities present in this study (median 2-3 visible parasites). Given that many of the adult eels were found to have swim bladder damage, concerns about their spawning migration through deeper oceanic environments can still be raised. To allow further investigations, we suggest that quantification of swim bladder damage should be implemented in eel-monitoring programs. Compared to other parasite pressure parameters, swim bladder damage provides additional information about past infections and future problems.

Extreme original data yield extreme decline effects.

Clements et al. respond to Munday's claim that his "reanalysis shows there is not an extreme decline effect in fish ocean acidification studies". They contend that extreme data reported in early studies authored by Dixson and Munday indeed result in an "extreme" decline effect in this field, and conclude that the decline effect is primarily driven by papers by particular authors.

The effects of ocean acidification on fishes - history and future outlook.

The effects of increased levels of carbon dioxide (CO2 ) on the Earth's temperature have been known since the end of the 19th century. It was long believed that the oceans' buffering capacity would counteract any effects of dissolved CO2 in marine environments, but during recent decades, many studies have reported detrimental effects of ocean acidification on aquatic organisms. The most prominent effects can be found within the field of behavioural ecology, e.g., complete reversal of predator avoidance behaviour in CO2 -exposed coral reef fish. Some of the studies have been very influential, receiving hundreds of citations over recent years. The results have also been conveyed to policymakers and publicized in prominent media outlets for the general public. Those extreme effects of ocean acidification on fish behaviour have, however, spurred controversy, given that more than a century of research suggests that there are few or no negative effects of elevated CO2 on fish physiology. This is due to sophisticated acid-base regulatory mechanisms that should enable their resilience to near-future increases in CO2 . In addition, an extreme "decline effect" has recently been shown in the literature regarding ocean acidification and fish behaviour, and independent research groups have been unable to replicate some of the most profound effects. Here, the author presents a brief historical overview on the effects of elevated CO2 and ocean acidification on fishes. This historical recap is warranted because earlier work, prior to a recent (c. 10 year) explosion in interest, is often overlooked in today's ocean acidification studies, despite its value to the field. Based on the historical data and the current knowledge status, the author suggests future strategies with the aim to improve research rigour and clarify the understanding of the effects of ocean acidification on fishes.

Meta-analysis reveals an extreme "decline effect" in the impacts of ocean acidification on fish behavior.

Ocean acidification-decreasing oceanic pH resulting from the uptake of excess atmospheric CO2-has the potential to affect marine life in the future. Among the possible consequences, a series of studies on coral reef fish suggested that the direct effects of acidification on fish behavior may be extreme and have broad ecological ramifications. Recent studies documenting a lack of effect of experimental ocean acidification on fish behavior, however, call this prediction into question. Indeed, the phenomenon of decreasing effect sizes over time is not uncommon and is typically referred to as the "decline effect." Here, we explore the consistency and robustness of scientific evidence over the past decade regarding direct effects of ocean acidification on fish behavior. Using a systematic review and meta-analysis of 91 studies empirically testing effects of ocean acidification on fish behavior, we provide quantitative evidence that the research to date on this topic is characterized by a decline effect, where large effects in initial studies have all but disappeared in subsequent studies over a decade. The decline effect in this field cannot be explained by 3 likely biological explanations, including increasing proportions of studies examining (1) cold-water species; (2) nonolfactory-associated behaviors; and (3) nonlarval life stages. Furthermore, the vast majority of studies with large effect sizes in this field tend to be characterized by low sample sizes, yet are published in high-impact journals and have a disproportionate influence on the field in terms of citations. We contend that ocean acidification has a negligible direct impact on fish behavior, and we advocate for improved approaches to minimize the potential for a decline effect in future avenues of research.

Changing epidemiology and age-specific incidence of cutaneous malignant melanoma in England: An analysis of the national cancer registration data by age, gender and anatomical site, 1981-2018.

BACKGROUND: The incidence of cutaneous malignant melanoma, which is mostly attributable (86%) to UV radiation exposure, has been steadily increasing over the past four decades in predominantly fair-skinned populations. Although public health campaigns are increasing sun-protective behaviour in England, their effect on melanoma incidence is largely unknown. We conducted a retrospective population-based cohort study to examine whether there have been changes in the epidemiology of melanoma in England during the past four decades. METHODS: Individual level data for patients diagnosed with melanoma in England during 1981-2018 were obtained from the Office for National Statistics/Public Health England. Average annual incidence rates were calculated by three age categories (0-34, 35-64, 65+ years), gender and anatomical site during the seven five-year time periods (1981-85 to 2011-15) and the recent three-year period (2016-18). The percentage change in incidence was calculated as change in the average incidence rate from the first (1981-85) to the last time period (2016-18). The Average Annual Percentage Change (AAPC) was estimated using the slope of the linear trend line fitted to the incidence rates by year of diagnosis. FINDINGS: During the 38-year period (1981-2018), a total of 265,302 cases of melanoma (45.7% males, 54.3% females) were registered in England. The average annual number of cases increased from 837/year in 1981-85 to 6963/year in 2016-18 in males (+732%), and from 1609/year in 1981-85 to 6952/year in 2016-18 in females (+332%). In the young age-group (0-34 years), the average annual incidence rates initially increased from 1981-85 to 2001-05 and then stabilised during the recent period (2006-18). In the middle age group (35-64 years), the rates increased by +332% (AAPC, 10.4%) in males (from 5.6/100,000 in 1981-85 to 24.2/100,000 in 2016-18) and +185% (AAPC, 5.7%) in females (from 10.2/100,000 in 1981-85 to 29.1/100,000 in 2016-18); and in the old age-group (65+ years) the rates increased by +842% (AAPC, 25.7%) in males (from 9.6/100,000 in 1981-85 to 90.4/100,000 in 2016-18) and +381% (AAPC, 11.2%) in females (from 12.5/100,000 in 1981-85 to 60.1/100,000 in 2016-18). The largest increase in incidence in both males and females was observed for melanoma of the trunk (+817%, AAPC, 24.8% in males and +613%, AAPC, 18.3% in females), followed by melanoma of upper limb (+750%, AAPC, 22.9% in males and 518%, AAPC, 15.5% in females). INTERPRETATION: It appears that the incidence of melanoma among young people in England has stabilised (or levelled off) in recent decades, whereas it continues to increase substantially in older population. These findings suggest that public health campaigns targeted at children/adolescents/parents may be favourably influencing melanoma incidence. The steeper increase in incidence in males is consistent with their relatively greater sun exposure and poor sun-protective behaviour. All the available evidence suggests that the enormous increase in the melanoma of the trunk and upper limb, since the 1980s, is most likely due to increasing trend in intermittent high intensity recreational UV radiation exposure (e.g. sunbathing, holidaying in places with strong sunlight, indoor tanning). FUNDING: This work was supported by Brighton and Sussex Medical School (BSMS).

Ocean acidification does not impair the behaviour of coral reef fishes.

The partial pressure of CO2 in the oceans has increased rapidly over the past century, driving ocean acidification and raising concern for the stability of marine ecosystems1-3. Coral reef fishes are predicted to be especially susceptible to end-of-century ocean acidification on the basis of several high-profile papers4,5 that have reported profound behavioural and sensory impairments-for example, complete attraction to the chemical cues of predators under conditions of ocean acidification. Here, we comprehensively and transparently show that-in contrast to previous studies-end-of-century ocean acidification levels have negligible effects on important behaviours of coral reef fishes, such as the avoidance of chemical cues from predators, fish activity levels and behavioural lateralization (left-right turning preference). Using data simulations, we additionally show that the large effect sizes and small within-group variances that have been reported in several previous studies are highly improbable. Together, our findings indicate that the reported effects of ocean acidification on the behaviour of coral reef fishes are not reproducible, suggesting that behavioural perturbations will not be a major consequence for coral reef fishes in high CO2 oceans.

On the Observation of Wild Zebrafish (Danio rerio) in India.

Zebrafish is one of the world's most widely used laboratory species, and it is utilized to answer important research questions in disparate fields such as biomedicine, genetics, developmental biology, pharmacology, toxicology, physiology, and evolution. Despite their popularity, very little is known about the biology of zebrafish in their natural habitat. This may, in part, be due to the difficulties associated with undertaking field trips to the remote areas of northern India, Nepal, and Bangladesh, which is the natural distribution range of zebrafish. Here, we present a field report describing a recent trip where we, together with local collaborators, visited several rivers in West Bengal, India, to observe wild zebrafish and their habitat. We present an overview of our observations on the biology of wild zebrafish, and the great variability of the different environments where they were found. We also include data collected on water chemistry parameters at 12 zebrafish sites, and weight data and photos of fish from these sites. We present extensive underwater videos of wild zebrafish and photographs of the sites, including video footage of courtship behavior. We show that the breeding period of wild zebrafish can be extended from the previous record of April-August to April-October. In addition, we provide practical advice for future zebrafish expeditions to this rural and inaccessible area. The goals of this article are to shed some light on the ecology of wild zebrafish, and to facilitate scientists in their future research trips. We hope that by observing zebrafish in the wild, we can increase our understanding of the natural ecology of this important model organism.

Brain cooling marginally increases acute upper thermal tolerance in Atlantic cod.

Physiological mechanisms determining thermal limits in fishes are debated but remain elusive. It has been hypothesised that motor function loss, observed as loss of equilibrium during acute warming, is due to direct thermal effects on brain neuronal function. To test this, we mounted cooling plates on the heads of Atlantic cod (Gadus morhua) and quantified whether local brain cooling increased whole-organism acute upper thermal tolerance. Brain cooling reduced brain temperature by 2-6°C below ambient water temperature and increased thermal tolerance by 0.5 and 0.6°C on average relative to instrumented and uninstrumented controls, respectively, suggesting that direct thermal effects on brain neurons may contribute to setting upper thermal limits in fish. However, the improvement in thermal tolerance with brain cooling was small relative to the difference in brain temperature, demonstrating that other mechanisms (e.g. failure of spinal and peripheral neurons, or muscle) may also contribute to controlling acute thermal tolerance.

Dental X-Rays and the Risk of Thyroid Cancer and Meningioma: A Systematic Review and Meta-Analysis of Current Epidemiological Evidence.

Background: Exposure to moderate-to-high doses of ionizing radiation is the only established environmental risk factor for thyroid cancer and brain and central nervous system tumors. Considering the high lifetime prevalence and frequency of exposure to dental X-rays, the most common source of diagnostic radiation exposure in the general population, even a small associated increase in cancer risk would be of considerable public health importance. With the objective to inform clinical practice and guidelines, we synthesized the current epidemiological evidence on the association between dental X-rays and the risk of thyroid cancer, meningioma, and other cancers of the head and neck region. Methods: The Medline, Embase, and Web of Science databases were searched to identify eligible studies. Summary odds ratio/relative risk estimates and confidence intervals were extracted, and pooled risk ratios (RRs) for each cancer were calculated using random effects meta-analysis. Results: The literature search identified 5537 publications; of these, 26 studies including 10,868 cancer patients were included in the synthesis. The random effects meta-analyses, based on seven studies of thyroid cancer (six case/control, one cohort) and eight studies of meningioma (all case/control), showed that multiple (or repeated) exposures to dental X-rays were significantly associated with an increased risk of thyroid cancer (pooled RR = 1.87 [95% confidence interval, CI 1.11-3.15]) and meningioma (pooled RR = 1.53 [CI 1.26-1.85]). There was no association with glioma, and there were too few studies of other cancers of the head and neck region to conduct a meaningful meta-analysis. Conclusions: Based on a meta-analysis of retrospective case/control studies, these findings provide some support to the hypothesis that multiple (or repeated) exposures to dental X-rays may be associated with an increased risk of thyroid cancer and meningioma. These studies did not include individual organ doses and ages at exposure, and are subject to recall bias and other limitations. Furthermore, the thyroid exposure has decreased dramatically over time from the use of thyroid shields and improved technology/equipment. Prospective studies, based on dental X-ray records and patient follow-up, are needed to test the hypothesis further and clarify the possible cancer risk associated with dental radiography, as although the risk at the individual level, particularly with improved technology/equipment, is likely to be very low, the proportion of the population exposed is high. Considering that about one-third of the general population in developed countries is routinely exposed to one or more dental X-rays per year, these findings manifest the need to reduce diagnostic radiation exposure as much as possible.

Are model organisms representative for climate change research? Testing thermal tolerance in wild and laboratory zebrafish populations.

Model organisms can be useful for studying climate change impacts, but it is unclear whether domestication to laboratory conditions has altered their thermal tolerance and therefore how representative of wild populations they are. Zebrafish in the wild live in fluctuating thermal environments that potentially reach harmful temperatures. In the laboratory, zebrafish have gone through four decades of domestication and adaptation to stable optimal temperatures with few thermal extremes. If maintaining thermal tolerance is costly or if genetic traits promoting laboratory fitness at optimal temperature differ from genetic traits for high thermal tolerance, the thermal tolerance of laboratory zebrafish could be hypothesized to be lower than that of wild zebrafish. Furthermore, very little is known about the thermal environment of wild zebrafish and how close to their thermal limits they live. Here, we compared the acute upper thermal tolerance (critical thermal maxima; CTmax) of wild zebrafish measured on-site in West Bengal, India, to zebrafish at three laboratory acclimation/domestication levels: wild-caught, F1 generation wild-caught and domesticated laboratory AB-WT line. We found that in the wild, CTmax increased with increasing site temperature. Yet at the warmest site, zebrafish lived very close to their thermal limit, suggesting that they may currently encounter lethal temperatures. In the laboratory, acclimation temperature appeared to have a stronger effect on CTmax than it did in the wild. The fish in the wild also had a 0.85-1.01°C lower CTmax compared to all laboratory populations. This difference between laboratory-held and wild populations shows that environmental conditions can affect zebrafish's thermal tolerance. However, there was no difference in CTmax between the laboratory-held populations regardless of the domestication duration. This suggests that thermal tolerance is maintained during domestication and highlights that experiments using domesticated laboratory-reared model species can be appropriate for addressing certain questions on thermal tolerance and global warming impacts.

Long-term acclimation to near-future ocean acidification has negligible effects on energetic attributes in a juvenile coral reef fish.

Increased levels of dissolved carbon dioxide (CO2) drive ocean acidification and have been predicted to increase the energy use of marine fishes via physiological and behavioural mechanisms. This notion is based on a theoretical framework suggesting that detrimental effects on energy use are caused by plasma acid-base disruption in response to hypercapnic acidosis, potentially in combination with a malfunction of the gamma aminobutyric acid type A (GABAA) receptors in the brain. However, the existing empirical evidence testing these effects primarily stems from studies that exposed fish to elevated CO2 for a few days and measured a small number of traits. We investigated a range of energetic traits in juvenile spiny chromis damselfish (Acanthochromis polyacanthus) over 3 months of acclimation to projected end-of-century CO2 levels (~ 1000 µatm). Somatic growth and otolith size and shape were unaffected by the CO2 treatment across 3 months of development in comparison with control fish (~ 420 µatm). Swimming activity during behavioural assays was initially higher in the elevated CO2 group, but this effect dissipated within ~ 25 min following handling. The transient higher activity of fish under elevated CO2 was not associated with a detectable difference in the rate of oxygen uptake nor was it mediated by GABAA neurotransmitter interference because treatment with a GABAA antagonist (gabazine) did not abolish the CO2 treatment effect. These findings contrast with several short-term studies by suggesting that end-of-century levels of CO2 may have negligible direct effects on the energetics of at least some species of fish.

Density differences between water masses preclude laminar flow in two-current choice flumes.

Two-current choice flumes are used to measure preference and avoidance behaviour in response to chemical cues in aquatic animals. If used correctly, they produce two parallel, non-overlapping, laminar water currents in which the animal can move freely and choose between the two currents. As climate change is affecting water temperature, and altered precipitation patterns are changing water salinity, two-current choice flumes are increasingly being used to test the choice between water currents of different temperatures and salinities. This inevitably means that water currents of different densities are being used simultaneously in the flume. Here, we investigated the tolerance range for density differences due to temperature and salinity in five common flume designs. Through dye tests and stepwise modifications of temperatures and salinities we determined the limits for laminar and non-overlapping flows. We also developed an automated method for quantifying the overlap precisely and objectively. The tolerance for density differences between the water currents where laminar and non-overlapping flows were maintained was surprisingly low, withstanding ± 0.5 °C temperature differences, and ± 0.1 PSU salinity differences, i.e. a maximum density difference of 0.28 gL-1. Above these very narrow limits we found a range where the flumes showed partly overlapping, stratified water currents that preclude easy determination of cue preference. We conclude that two-current choice flumes are not suitable for testing the behavioural choices of aquatic animals using water currents of anything other than minor differences in temperature and/or salinity.

Behavioural alterations induced by the anxiolytic pollutant oxazepam are reversible after depuration in a freshwater fish.

Anthropogenic pharmaceutical pollutants have been detected in nature across the globe, and recent work has shown negative effects of pharmaceuticals on the health and welfare of many animals. However, whether alterations can be reversed has been poorly investigated, although such studies are essential to assess the effects of high-peak exposure events in waterways where pharmaceutical concentrations are usually low. In this study, we investigated the effects of two concentrations (environmentally relevant 1 µg L-1 and high 100 µg L-1) of oxazepam, an anxiolytic commonly detected in aquatic environments, and whether behavioural alterations are reversible after depuration. Specifically, we measured daytime and night-time swimming activity and daytime behaviours related to boldness (foraging, sheltering and routine swimming activity) using the freshwater burbot (Lota lota). We found that both swimming activity and boldness were affected by oxazepam. Fish exposed to the higher level had a higher burst swimming duration (i.e., fast swimming bouts), both in the daytime and night-time trials. Further, fish exposed to the lower oxazepam level spent less time sheltering than control- and high-level exposed fish, but there was no difference between the control and high oxazepam treatments. For routine swimming activity, quantified in the boldness trials, and for latency to forage, there were no treatment effects. When retesting the fish after depuration, the detected behavioural alterations were no longer present. Since the magnitude of these effects were not consistent across endpoints, our study suggests that oxazepam might not be a great concern for this particular, stress tolerant, species, highlighting the importance of evaluating species-specific effects of pharmaceuticals. The observation that the effects we did find were reversible after depuration is encouraging, and indicates that rapid restoration of behaviours after removal from oxazepam contamination is possible.

Direct and indirect effects of chemical contaminants on the behaviour, ecology and evolution of wildlife.

Chemical contaminants (e.g. metals, pesticides, pharmaceuticals) are changing ecosystems via effects on wildlife. Indeed, recent work explicitly performed under environmentally realistic conditions reveals that chemical contaminants can have both direct and indirect effects at multiple levels of organization by influencing animal behaviour. Altered behaviour reflects multiple physiological changes and links individual- to population-level processes, thereby representing a sensitive tool for holistically assessing impacts of environmentally relevant contaminant concentrations. Here, we show that even if direct effects of contaminants on behavioural responses are reasonably well documented, there are significant knowledge gaps in understanding both the plasticity (i.e. individual variation) and evolution of contaminant-induced behavioural changes. We explore implications of multi-level processes by developing a conceptual framework that integrates direct and indirect effects on behaviour under environmentally realistic contexts. Our framework illustrates how sublethal behavioural effects of contaminants can be both negative and positive, varying dynamically within the same individuals and populations. This is because linkages within communities will act indirectly to alter and even magnify contaminant-induced effects. Given the increasing pressure on wildlife and ecosystems from chemical pollution, we argue there is a need to incorporate existing knowledge in ecology and evolution to improve ecological hazard and risk assessments.

Exposure to elevated carbon dioxide does not impair short-term swimming behaviour or shelter-seeking in a predatory coral-reef fish.

Adult bluespotted rockcod Cephalopholis cyanostigma, a coral-reef grouper, were acclimated to either ambient (mean ± s.d. 406 ± 21 µatm; 1 atmos = 101325 Pa) or high pCO2 (945 ± 116 µatm) conditions in a laboratory for 8-9 days, then released at the water surface directly above a reef (depth c. 5 m) and followed on video camera (for 191 ± 21 s) by scuba divers until they sought cover in the reef. No differences were detected between groups in any of the six measured variables, which included the time fish spent immobile after release, tail beat frequency during swimming and the time required to locate and enter the protective shelter of the reef.