Morten Bo Søndergaard Svendsen

Design and setup of intermittent‐flow respirometry system for aquatic organisms

Intermittent-flow respirometry is an experimental protocol for measuring oxygen consumption in aquatic organisms that utilizes the best features of closed (stop-flow) and flow-through respirometry while eliminating (or at least reducing) some of their inherent problems. By interspersing short periods of closed-chamber oxygen consumption measurements with regular flush periods, accurate oxygen uptake rate measurements can be made without the accumulation of waste products, particularly carbon dioxide, which may confound results. Automating the procedure with easily available hardware and software further reduces error by allowing many measurements to be made over long periods thereby minimizing animal stress due to acclimation issues. This paper describes some of the fundamental principles that need to be considered when designing and carrying out automated intermittent-flow respirometry (e.g. chamber size, flush rate, flush time, chamber mixing, measurement periods and temperature control). Finally, recent advances in oxygen probe technology and open source automation software will be discussed in the context of assembling relatively low cost and reliable measurement systems.

Conservation Physiology of Marine Fishes: State of the Art and Prospects for Policy

The state of the art of research on the environmental physiology of marine fishes is reviewed from the perspective of how it can contribute to conservation of biodiversity and fishery resources. A major constraint to application of physiological knowledge for conservation of marine fishes is the limited knowledge base; international collaboration is needed to study the environmental physiology of a wider range of species. Multifactorial field and laboratory studies on biomarkers hold promise to relate ecophysiology directly to habitat quality and population status. The ‘Fry paradigm’ could have broad applications for conservation physiology research if it provides a universal mechanism to link physiological function with ecological performance and population dynamics of fishes, through effects of abiotic conditions on aerobic metabolic scope. The available data indicate, however, that the paradigm is not universal, so further research is required on a wide diversity of species. Fish physiologists should interact closely with researchers developing ecological models, in order to investigate how integrating physiological information improves confidence in projecting effects of global change; for example, with mechanistic models that define habitat suitability based upon potential for aerobic scope or outputs of a dynamic energy budget. One major challenge to upscaling from physiology of individuals to the level of species and communities is incorporating intraspecific variation, which could be a crucial component of species’ resilience to global change. Understanding what fishes do in the wild is also a challenge, but techniques of biotelemetry and biologging are providing novel information towards effective conservation. Overall, fish physiologists must strive to render research outputs more applicable to management and decision-making. There are various potential avenues for information flow, in the shorter term directly through biomarker studies and in the longer term by collaborating with modellers and fishery biologists.

Sources of variation in oxygen consumption of aquatic animals demonstrated by simulated constant oxygen consumption and respirometers of different sizes

As intermittent-flow respirometry has become a common method for the determination of resting metabolism or standard metabolic rate (SMR), this study investigated how much of the variability seen in the experiments was due to measurement error. Experiments simulated different constant oxygen consumption rates (M˙O2 ) of a fish, by continuously injecting anoxic water into a respirometer, altering the injection rate to correct for the washout error. The effect of respirometer-to-fish volume ratio (RFR) on SMR measurement and variability was also investigated, using the simulated constant M˙O2 and the M˙O2 of seven roach Rutilus rutilus in respirometers of two different sizes. The results show that higher RFR increases measurement variability but does not change the mean SMR established using a double Gaussian fit. Further, the study demonstrates that the variation observed when determining oxygen consumption rates of fishes in systems with reasonable RFRs mainly comes from the animal, not from the measuring equipment.

Simulation-based training for colonoscopy: establishing criteria for competency.

AbstractThe aim of this study was to create simulation-based tests with credible pass/fail standards for 2 different fidelities of colonoscopy models.Only competent practitioners should perform colonoscopy. Reliable and valid simulation-based tests could be used to establish basic competency in colonoscopy before practicing on patients.Twenty-five physicians (10 consultants with endoscopic experience and 15 fellows with very little endoscopic experience) were tested on 2 different simulator models: a virtual-reality simulator and a physical model. Tests were repeated twice on each simulator model. Metrics with discriminatory ability were identified for both modalities and reliability was determined. The contrasting-groups method was used to create pass/fail standards and the consequences of these were explored.The consultants significantly performed faster and scored higher than the fellows on both the models (P < 0.001). Reliability analysis showed Cronbach &agr; = 0.80 and 0.87 for the virtual-reality and the physical model, respectively. The established pass/fail standards failed one of the consultants (virtual-reality simulator) and allowed one fellow to pass (physical model).The 2 tested simulations-based modalities provided reliable and valid assessments of competence in colonoscopy and credible pass/fail standards were established for both the tests. We propose to use these standards in simulation-based training programs before proceeding to supervised training on patients.

Not So Fast: Swimming Behavior of Sailfish during Predator–Prey Interactions using High-Speed Video and Accelerometry

Billfishes are considered among the fastest swimmers in the oceans. Despite early estimates of extremely high speeds, more recent work showed that these predators (e.g., blue marlin) spend most of their time swimming slowly, rarely exceeding 2 m s(-1). Predator-prey interactions provide a context within which one may expect maximal speeds both by predators and prey. Beyond speed, however, an important component determining the outcome of predator-prey encounters is unsteady swimming (i.e., turning and accelerating). Although large predators are faster than their small prey, the latter show higher performance in unsteady swimming. To contrast the evading behaviors of their highly maneuverable prey, sailfish and other large aquatic predators possess morphological adaptations, such as elongated bills, which can be moved more rapidly than the whole body itself, facilitating capture of the prey. Therefore, it is an open question whether such supposedly very fast swimmers do use high-speed bursts when feeding on evasive prey, in addition to using their bill for slashing prey. Here, we measured the swimming behavior of sailfish by using high-frequency accelerometry and high-speed video observations during predator-prey interactions. These measurements allowed analyses of tail beat frequencies to estimate swimming speeds. Our results suggest that sailfish burst at speeds of about 7 m s(-1) and do not exceed swimming speeds of 10 m s(-1) during predator-prey interactions. These speeds are much lower than previous estimates. In addition, the oscillations of the bill during swimming with, and without, extension of the dorsal fin (i.e., the sail) were measured. We suggest that extension of the dorsal fin may allow sailfish to improve the control of the bill and minimize its yaw, hence preventing disturbance of the prey. Therefore, sailfish, like other large predators, may rely mainly on accuracy of movement and the use of the extensions of their bodies, rather than resorting to top speeds when hunting evasive prey.

Laser Speckle Contrast Imaging for Monitoring Changes in Microvascular Blood Flow

Background/Aims: Microvascular blood flow is essential for healing and predicts surgical outcome. The aim of the current study was to investigate the relation between fluxes measured with the laser speckle contrast imaging (LSCI) technique and changes in absolute blood flow. In addition, we studied the reproducibility of the LSCI technique when assessing the intra-abdominal microcirculation of the pig. Methods: During trial 1, a fish gill arch was mechanically perfused with heparinized fish blood under controlled stepwise-altered flow rates alongside mechanically induced movement artefacts. The microcirculation of the fish gill was simultaneously assessed with the LSCI technique. In trial 2, microcirculation was measured in the stomach, liver, and small intestine of 10 pigs by two observers. Results: A linear correlation was observed between flux and volumetric flow. During conditions of no volumetric flow, the high recording speed with the LSCI technique registered the movement artefacts as flow signals. The LSCI measurements showed good correlation and agreement between the two observers when assessing microcirculation in the stomach, liver, and small intestine (r2 = 0.857, 0.956, and 0.946; coefficients of variation = 6.0, 3.2, and 6.4%, respectively). Conclusion: Due to the non-contact and real-time assessment over large areas, LSCI is a promising technique for the intraoperative assessment of intra-abdominal microcirculation. A linear correlation between flux and volumetric flow was found, in accordance with previous experimental studies. However, movement artefacts affect flux measurements, and the choice of the sampling speed must be made with care, depending on the given setting.

Effect of closed v. intermittent‐flow respirometry on hypoxia tolerance in the shiner perch Cymatogaster aggregata

This study compares the critical oxygen saturation (O2 crit ) levels of the shiner perch Cymatogaster aggregata obtained using two different methods wherein hypoxia is induced either by the fishs respiration (closed respirometry) or by degassing oxygen with nitrogen (intermittent-flow respirometry). Fish exhibited loss of equilibrium at a higher O2 saturation in the closed respirometry method when compared with the intermittent-flow method. Utilization of closed respirometry yielded O2 crit measurements that were almost twice as high as those obtained with intermittent-flow respirometry. The lower hypoxia tolerance in closed respirometry is consistent with additional stress, caused by a build-up of ammonia and carbon dioxide and a faster rate in dissolved oxygen decline. The results indicate that these two methods of determining hypoxia tolerance in aquatic organisms are not comparable, and that much care should be given to method choice.

Fast-starting after a breath: air-breathing motions are kinematically similar to escape responses in the catfish Hoplosternum littorale

ABSTRACT Fast-starts are brief accelerations commonly observed in fish within the context of predator–prey interactions. In typical C-start escape responses, fish react to a threatening stimulus by bending their body into a C-shape during the first muscle contraction (i.e. stage 1) which provides a sudden acceleration away from the stimulus. Recently, similar C-starts have been recorded in fish aiming at a prey. Little is known about C-starts outside the context of predator–prey interactions, though recent work has shown that escape response can also be induced by high temperature. Here, we test the hypothesis that air-breathing fish may use C-starts in the context of gulping air at the surface. Hoplosternum littorale is an air-breathing freshwater catfish found in South America. Field video observations reveal that their air-breathing behaviour consists of air-gulping at the surface, followed by a fast turn which re-directs the fish towards the bottom. Using high-speed video in the laboratory, we compared the kinematics of the turn immediately following air-gulping performed by H. littorale in normoxia with those of mechanically-triggered C-start escape responses and with routine (i.e. spontaneous) turns. Our results show that air-breathing events overlap considerably with escape responses with a large stage 1 angle in terms of turning rates, distance covered and the relationship between these rates. Therefore, these two behaviours can be considered kinematically comparable, suggesting that air-breathing in this species is followed by escape-like C-start motions, presumably to minimise time at the surface and exposure to avian predators. These findings show that C-starts can occur in a variety of contexts in which fish may need to get away from areas of potential danger.

Plasma osmolality and oxygen consumption of perchPerca fluviatilisin response to different salinities and temperatures: osmoregulation and MṀo2inp. fluviatilis

The present study determined the blood plasma osmolality and oxygen consumption of the perch Perca fluviatilis at different salinities (0, 10 and 15) and temperatures (5, 10 and 20° C). Blood plasma osmolality increased with salinity at all temperatures. Standard metabolic rate (SMR) increased with salinity at 10 and 20° C. Maximum metabolic rate (MMR) and aerobic scope was lowest at salinity of 15 at 5° C, yet at 20° C, they were lowest at a salinity of 0. A cost of osmoregulation (SMR at a salinity of 0 and 15 compared with SMR at a salinity of 10) could only be detected at a salinity of 15 at 20° C, where it was 28%. The results show that P. fluviatilis have capacity to osmoregulate in hyper-osmotic environments. This contradicts previous studies and indicates intraspecific variability in osmoregulatory capabilities among P. fluviatilis populations or habitat origins. An apparent cost of osmoregulation (28%) at a salinity of 15 at 20° C indicates that the cost of osmoregulation in P. fluviatilis increases with temperature under hyperosmotic conditions and a power analysis showed that the cost of osmoregulation could be lower than 12·5% under other environmental conditions. The effect of salinity on MMR is possibly due to a reduction in gill permeability, initiated to reduce osmotic stress. An interaction between salinity and temperature on aerobic scope shows that high salinity habitats are energetically beneficial during warm periods (summer), whereas low salinity habitats are energetically beneficial during cold periods (winter). It is suggested, therefore, that the seasonal migrations of P. fluviatilis between brackish and fresh water is to select an environment that is optimal for metabolism and aerobic scope.

A reduced gastric corpus microvascular blood flow during Ivor-Lewis esophagectomy detected by laser speckle contrast imaging technique

Abstract Background: Reduced microvascular blood flow is related to anastomotic insufficiency following esophagectomy, emphasizing a need for intraoperative monitoring of the microcirculation. This study evaluated if laser speckle contrast imaging (LSCI) was able to detect intraoperative changes in gastric microcirculation. Methods: Gastric microcirculation was assessed prior to and after reconstruction of gastric continuity in 25 consecutive patients operated for adenocarcinoma with open Ivor-Lewis esophagectomy while hemodynamic variables were recorded. Results: During upper laparotomy, microcirculation at the corpus decreased by 25% from baseline to mobilization of the stomach (p = .008) and decreased further (to a total decrease of 40%) following gastric pull to the thorax (p = .013). On the other hand, microcirculation at the antrum did not change significantly after gastric mobilization (p = .091). The decrease in corpus microcirculation took place unrelated to central cardiovascular variables. Conclusion: Using LSCI technique, we identified a reduced microcirculation at the corpus area during open Ivor-Lewis esophagectomy. LSCI provides an option for real-time assessment of gastric microcirculation and could form basis for intraoperative stabilization of the microcirculation.