Reflex impairment and vitality in white sturgeon exposed to simulated capture stressors

White sturgeon, NEEF 2016

A study (McLean et al. 2016) of reflex impairment in white sturgeon exposed to sustained exercise and elevated temperature showed whole-animal stress responses to simulated capture. The RAMP impairment index (a simple proportion of measured reflex actions that were impaired) was used to quantify relationships between treatment times, recovery times, and RAMP score.

The upper figure shows increasing RAMP score with increasing exercise (minutes) in summer (filled circle) and winter (filled triangle) temperatures. The lower figure shows increasing recovery time with increasing RAMP score in summer and winter temperatures. Figures adapted from McLean et al. 2016.

The authors state: “Our study demonstrates that reflex impairment (RAMP) indices are a promising tool to predict post-release vitality in white sturgeon exposed to acute fisheries encounters, such as an angling event. The reflexes used in our RAMP protocol were chosen so that multiple neurological and/or muscle pathways underlying the overall stress response were tested. What we found was that sturgeon exposed to fishing-related stressors had higher RAMP scores and took significantly longer to recover than control fish. The relationship between reflex impairment and stressor intensity (i.e. fishery-related treatment) indicates that sturgeon are undergoing whole-animal (or tertiary) responses to varying degrees of capture stress. Reflex impairment indicators were surprisingly sensitive to fisheries stressors. Control fish had all reflexes intact, whereas multiple reflexes were absent after fish were treated.

It is important to note that it was not the aim of this study to produce accurate mortality estimates for use in C&R fisheries, but rather to explore the use of RAMP on a sturgeon species frequently angled in the wild. We recognize the subjectivity of a whole-animal assessment and categorization; however, given the statistically significant difference in RAMP scores of observationally ‘recovered’ and ‘unrecovered’ sturgeon, we suggest that RAMP is an effective tool for predicting a lowered state of vitality post-release and that this suggests a continuum to an increased risk of delayed mortality.”

RAMP method video developed by ILVO

ILVO (Belgium Institute for Agricultural and Fisheries Research) has developed RAMP methods for three species of flatfish (plaice, sole, and dab) in European fisheries.

The first video sets the scene and explains the potential relevance of this method in relation to the recently reformed European Common Fisheries Policy.

The second video explains and demonstrates reflex tests in more detail and may guide other investigators in defining and recognizing reflex actions.

An excerpt from the video text explains, “A staggering amount of commercially-caught fish is being thrown overboard. Some say that all of those discarded fish are either dead before they hit the water or they die soon after, victims of predation or injury. But others argue that some of those species are strong enough to survive after being discarded and live long enough to reproduce. The European Common Fisheries Policy was recently reformed and will now phase in a ban on discarding, meaning that fishers will have to land everything they catch. The idea behind the ban is to stimulate more selective fishing techniques, because it will be in the fisher’s interest to only catch the most valuable fish. However, by landing everything, this ban risks killing more fish than before. If a juvenile fish lives long enough after being discarded to spawn new fish, it should be given that opportunity. For this reason, the discard policy provides an important exception: if a certain species can be scientifically proven to have a high chance of survival, fish of that species should be thrown back after catch. Researchers at the Institute for Agricultural and Fisheries Research (or ILVO) in Ostend, Belgium are testing the most commercially important species of flatfish - plaice, sole and dab – for their likelihood of survival.”

Questions and answers about observer bias in RAMP

Refer to the previous post for background on observer bias in RAMP.

Q: What are the options when grappling with cognitive/expectation and sampling biases in manipulative fisheries research experiments under sometimes challenging conditions at sea?

A: Begin by training and calibrating observation. We all recognize vitality when we see animals with high vitality. This recognition is based on rapid visual assimilation of information about several traits including injury, activity, and responsiveness. We cannot separate our cognitive impression of vitality level from the act of observing individual traits and scoring their presence or absence. Presence or absence of reflex actions is scored relative to control animals which have a set of reflex actions consistently present. Reflex actions range from clearly seen through weakening stages to clearly absent. As the animal becomes more stressed and impairment increases, the interaction of impression and scoring observations contributes bias. 

If observers are trained to clearly recognize a suite of real reflex actions in the species of interest, then correctly recognizing the impairment or absence of those reflex actions should be a realistic accomplishment. An experiment to test for the effect of observer bias and variability in scoring reflex actions could be conducted in the lab or field if enough fish and observers are available. Stress some fish (air exposure) to produce replicates over a range of RAMP impairment scores and have the observers sample reflex actions. Blind the study treatments from observers. Estimates for observer bias from stress studies with different species will be useful for improving observer training by identifying protocols that need to be more defined and less subject to observer opinions. Alternatively, Benoît et al. (2010) modeled observer bias as a random factor. 

Q: How can we achieve a blinded experimental design if the experimenter who assigns or is aware of experimental treatments also scores reflex impairment on board (commercial) vessels?

A: Perform some fish experiments on observer bias outlined above and decide how important observer bias is after training with well-defined protocols for testing individual reflex actions. The bias problem may be mitigated by training using clear definitions of present or absent for reflex actions. I will assume that the vessel captain is conducting the experimental fishing treatments. So the captain could be given treatment conditions by the scientist and then could conduct fishing by assigning treatments randomly without the knowledge of the scientist observer. However tow time, soak time, or haul time and catch volume will be apparent to observers. 

Q: Is an observer influenced in his/her ability to score reflexes if, apart from knowing the treatment, also the condition of an organism is evident even before the scoring begins? Is there any option to minimise this?

A: We cannot separate the correlation between overall impression of vitality and scoring reflex actions. However, we can be trained to clearly recognize the presence of reflex actions. Any impairment through weakness, delay, or loss of action is scored absent.  The key method for minimizing observer bias for reflex actions is to clearly establish what the suite of reflex actions look like when they are consistently present in control animals. If presence of a reflex action is difficult or inconsistent to determine then it is not a good candidate for testing. Any deviation from control appearance in action strength or delayed time for action can be considered impaired and scored absent. The goal is to eliminate variability in detection of presence for reflex actions. By sharpening the decision criteria, bias and variability can be reduced. This idea can be tested using the outlined experiment design.

Q: Seeing that vitality assessments of discarded fish in Europe are now being developed in several places is there a need to also quantitatively evaluate the ability of different observers to score reflexes consistently? What would be the best setup for such a training exercise? 

A: As mentioned above, a stress experiment can be conducted to quantify observer bias and consistency.  With enough replicate fish and observers, an air stress experiment could produce fish with varying levels of reflex action impairment. These fish could be sampled by observers with defined criteria and using an experimental design for testing the effects of observer variability and bias. The effect of training could also be evaluated using this design.

Survival of schooling small pelagic fish discarded from purse seine fisheries

Greenback horse mackerel, Trachurus declivis 

Purse seine fishing

Vitality impairment and RAMP can be used to determine survival of discarded schooling small pelagic fish in purse seine fisheries.  When catch is too large, fish are “slipped” for the net and discarded. These discarded fish are usually exposed to some level of hypoxic conditions associated with crowding in the purse seine. Elevated temperature in surface water may be a stressor. Skin abrasion and scale loss can occur in the net. Many small pelagic species (mackerel, sardine, anchovy, smelt, herring) caught in purse seines are obligate or facultative schoolers that reflexively form groups mediated by the optomotor response. Vitality impairment can be tested for individual fish or groups. See Davis and Ottmar, 2006 for testing groups of free-swimming fish. Schooling fish seek the company of species mates, so testing groups of schooling fish is probably the most informative method. How is this testing done and linked with delayed mortality in captive observation tanks?    

RAMP links vitality impairment scores with delayed mortality scores. The RAMP estimate for delayed mortality is only as good as the mortality estimates from captive observation or tagging experiments. How many replicates are needed in captive observation experiments? The purse seine schooling species need to be held in groups. A replicate group size of ten fish is good for schooling. These fish must be held in good water quality and circulation, in a circular tank size that allows schooling. For initial RAMP formulation, you will need 10 replicate groups of ten fish each. Observations of reflex action impairment and delayed mortality should be made over a range of stressor intensities that result in delayed mortality of 0 to 100%. Then replicate vitality impairment scores are linked with replicate delayed mortality scores to form the RAMP which can be validated with further experiments and replication.

Suuronen, 2005  Stressors in capture and escape of fisheries.

Fish can be sampled from any point on the fishing process, depending on the stressors of interest. Reflex action testing can be made on a group of fish held in a circular observation tank big enough for schooling (See Davis and Ottmar, 2006).  Possible reflex actions for testing include: orientation; schooling; rheotaxis; startle response to sound or light; swimming to bottom of tank. Injuries can also be noted; abrasion, scale loss. After testing the replicate group can then be placed in a holding tank and monitored for delayed mortality through five to ten days. 

Herring lose schooling, orientation, and tail beat frequency increases as the purse seine is drawn smaller (Morgan, 2014). Fatigue and hypoxia are possible stressors in purse seines (Tenningen 2014).

For discard species caught in purse seines that are not schooling fish, or are larger schooling fish, individual fish can be tested for vitality impairment.  Reflex actions tested can include: body flex, orientation, eye roll, operculum or mouth clamp, tail grab, righting, startle.  These fish can be tagged for identification and held together for five to ten days in tanks to determine delayed mortality.

Flexibility of using RAMP to determine bycatch mortality rates for Tanner crab caught in Alaska bottom trawls

Tanner crab (Chionoecetes bairdi), AFSC

Yochum et al. 2015 evaluated the flexibility of RAMP methodology by creating a RAMP for Tanner crab (Chionoecetes bairdi) discarded from the groundfish bottom trawl fishery in the Gulf of Alaska and comparing it to a previously established RAMP for unobserved Tanner crab bycatch (encountered gear and remained on the seafloor) from the bottom trawl fishery in the Bering Sea. The authors found that: “The two RAMPs and the overall mortality rates calculated using these predictors were comparable. However, we detected significant differences between RAMPs. While probabilities of mortality were similar between the two studies for crab with all or no reflexes missing, discarded crab with intermediate reflex impairment had lower mortality probabilities than those from the unobserved-bycatch study. Our results indicate that a RAMP may produce more accurate mortality estimates when applied to animals experiencing similar stressors as those evaluated to create the RAMP, through similar methodology.”

Conditions for holding crab after exposure to stressors can control delayed mortality and should be considered in experiments. “There were differential mortality rates by holding type. Higher mortality rates occurred in the on-board tanks (where the crab were held for the first few days) and in the laboratory tank. Moreover, Score-zero crab died in the holding tanks, but not in the at-sea cages. These results indicate that holding tanks contribute additional stressors, either due to transport, additional handling, or stress from being held in an unnatural setting or at temperatures greater than what was experienced in their natural environment. 

Our holding duration of two weeks was sufficient to determine mortality for all Scores. Given that it can take longer for Score-zero animals to die than those with higher Scores, our holding period allowed us to sufficiently capture Score-zero mortalities. However, the death of a Score-zero crab at day 12 may indicate that holding for more than a week confuses mortality attributed to fishing stressors with that from captivity.”

Evaluation of RAMP flexibility was made by comparing results from different studies. “To evaluate the divergence between the RAMPs we analyzed the differences between the studies. The primary difference was in experimental methods, namely the treatment of the crab before assessment. Crab from the Discard-mortality study were exposed to air for 90min on average (range from 9 to 230min) without any “recovery” in water. In contrast, crab from the Unobserved-mortality study had only brief air exposure and were held in water while awaiting assessment (generally less than 15 min), which may have allowed some recovery. 

These differences in air exposure and recovery in water probably affected the relationship between observed reflex impairments and delayed mortality and hence accounted for the discrepancy between RAMPs. Prolonged air exposure and experiencing cold temperatures was linked with increased delayed and instant mortality, number of autonomies for crab, as well as reduced vigor, juvenile growth, and feeding rates (Carls and O’Clair, 1995; Giomi et al., 2008; Grant, 2003; Stoner, 2009; Warrenchuk and Shirley, 2002). Stoner (2009) found that reflex impairment score and exposure to freezing temperatures were nearly linearly related for Tanner crab. Moreover, he found that the different RAMP reflexes had variable sensitivity to freezing temperatures, namely that the chela closure reflex was the most sensitive reflex, and mouth closure was least. Similarly, Van Tamelen (2005) found that the legs and eyes of snow crab cooled faster than the body, perhaps making them more susceptible to cold air exposure. We hypothesize that the prolonged air exposure for the Discard-mortality study likely impaired the crabs’ reflexes and resulted in higher Scores.”

Recommendations made by the authors. “Results from this study indicate that bias can be introduced in mortality rate estimates when using a RAMP created for one study to estimate mortality rates for a different study where the experimental methods differ, especially with respect to air exposure and recovery in water before assessment. However, when RAMP is used only to approximate mortality rates or to make comparisons between gear types or uses, a previously established RAMP could be used with caution, especially if animals with intermediate Scores are not predominant. For more accurate bycatch mortality rate estimates, our results indicate the importance of using a RAMP that was created by assessing animals that experienced similar stressors to those which the RAMP will be applied. Namely, the procedure for assessing the animals should be similar. We feel that the amount of time the animal spends out of water before assessment be standardized within a time range, along with whether or not the animal is allowed to recover in water before assessment, unless these variables are the treatments being studied.” 

“Our results indicate that consistency in methodology and relevance with respect to mimicking actual fishing stresses for the RAMP approach increases the flexibility of RAMP. It is therefore important, when creating a RAMP, to create repeatable methods that are well documented when publishing. RAMP reflexes should be assessed in a specified order to prevent bias from reflexes that are physiologically linked. If there is a reflex that influences the determination of other reflexes it should be assessed last or not at all. Reflexes that are difficult to determine presence or absence should not be used, and it should be clear in the methods what constitutes an “absent” reflex and how immediate mortalities are treated (are they given a Score or classified separately?). In addition, when a RAMP is being created, data should be recorded on all possible stressors, including injury, and evaluated for their contribution to mortality. Moreover, effort should be made (within the logistical constraints of field and laboratory research) to minimize additional stressors that are unrelated to the fishing stressors of interest.”

Sublethal effects of simulated angling capture (fatigue and air exposure) in snapper: reflex impairment and physiological stress.

Snapper, Pagrus auratus. Floor Anthoni 2006.

A study of reflex impairment and physiological stress was conducted with captured snapper and published in JEMBE 2014 by McArley, T.J & Herbert, N.A.  Fish were exposed to simulated angling by chasing to fatigue, followed by air exposure.  The authors' text is quoted below.

Reflex impairment (RI) was measured for seven reflex actions (Table 1). “If the presence of a positive reflex response was ambiguous it was scored as absent. The entire RI assessment was completed in less than 50 s, of which the fish was exposed to air for approximately 30 s. Reflexes were scored present (1) or absent (0) for individual fish and the RI score (proportion of reflexes impaired) was calculated by dividing the number of reflexes absent by the total number of possible reflexes (Davis, 2010). For example if four out of the seven possible reflexes were absent a fish would be given an RI score of 0.57.”

Reflex impairment is a potential measure of vitality loss after exposure of snapper to angling stressors: 

“A primary aim of this study was to assess the potential use of RI as a simple tool for measurement of fish vitality following angling and our lab-based trials indicate RI has the potential to be used in this way. RI was significantly related to the duration of strenuous exercise and air exposure (Fig. 1) and therefore provided a good index of fish condition. Importantly, fish exposed to more severe stress treatments exhibited greater RI than those exposed to more mild stress treatments, a finding that agrees with several other studies of RI in fish (Barkley and Cadrin, 2012; Brownscombe et al., 2013; Campbell et al., 2010b; Davis, 2007; Humborstad et al., 2009; Raby et al., 2012, 2013). Furthermore, greater RI was associated with significantly higher plasma lactate concentration and reduced muscle pH suggesting that RI can indicate (predict) an alteration in physiological condition.” 

Anaerobic respiration is associated with lactate production and reflex impairment:

“Burst swimming is powered by anaerobic respiration fueled by stored energy in white muscle (Milligan, 1996) and the lactic acid produced accumulates rapidly in muscle tissue and then “spills over” into circulation after a 5–10 min delay (Wood, 1991). Plasma lactate therefore serves as a useful indicator of anaerobic respiration in fish (Gale et al., 2011; Lowe and Wells, 1996; Meka and McCormick, 2005) and, as fish performed burst swimming during chasing, it is unsurprising that plasma lactate correlated positively with the duration of chasing stress and that both muscle pH and blood pH were lower in fish chased for longer periods. Physiological alterations appeared to be more pronounced in summer than in winter suggesting that when water temperature is warmer a more severe stress response appears to occur in snapper.”

Physiological basis for reflex impairment: 

“RI is thought to have a physiological basis (Davis, 2010) and a significant relationship between RI and increased plasma lactate concentration has been observed in salmonids (Raby et al., 2013). As RI is measured directly after stressor exposure physiological disturbances that manifest quickly during stress are likely causes of RI. Physiological alterations such as cortisol concentration that can plateau 30 min - 1 h after the initial stressor exposure (Milligan, 1996; Wendelaar Bonga, 1997) are therefore unlikely to be directly responsible for the RI measured in this study. In this study most RI occurred as a result of an inability to perform reflexes involving powerful muscular contractions, such as the gag reflex, body flexing and the startle response. Powerful muscular contraction is fueled by anaerobic metabolism in white muscle fibres and can only be maintained for short periods (Milligan, 1996). As higher RI scores were correlated with lower muscle pH and higher concentrations of plasma lactate it is hypothesized that muscle fatigue resulting from anaerobic metabolism performed during strenuous exercise caused the majority of the observed RI. The muscle pH and plasma lactate concentrations associated with the same RI scores, however, were different in summer and winter (Fig. 4) and there was no difference between the summer and winter measures of fish vitality (RI) and mortality. This suggests that rather than being causes of RI, plasma lactate concentration and muscle pH may have been indicators of an unmeasured physiological process that impaired some of the reflexes quantified in the current study. Other reflexes we measured that were not as commonly impaired, such as the righting response and vestibular ocular, are essentially neurological and their impairment likely results from alternative pathways to those measured in this study.”

Mortality was rarely observed when snapper were exposed to angling conditions: 

“Despite the limitations of comparing our mortality estimates to real fishing scenarios the findings provide evidence that strenuous exercise and air exposure imposed during angling, are not likely to be direct causes of discard mortality in P. auratus. During the collection of sub-legal snapper from the wild for this study, fish were landed relatively quickly (approximately 15 to 30 s), and were typically unhooked in less than 30 s. In investigations of authentic angling events for P. auratus of comparable size (< 270 mm FL) in south eastern Australia, the majority of fish were landed in less than 30 s (Broadhurst et al., 2012; Grixti et al., 2010) and had less than 30 second exposure to air (Broadhurst et al., 2012). Thus, the 5 minute strenuous exercise period and the 3 minute air exposure period in this study must be considered extreme levels for recreational angling and probably rarely occur in authentic angling events. Encouragingly, even with these high levels of stress, little mortality was seen against a backdrop of high summer water temperatures.”

Predation risk for snapper that show reflex impairment associated with angling: 

“It is often overlooked in catch and release studies but a potentially important contributor to discard mortality is post-release predation (Raby et al., 2014). While no measure of predation risk was assessed in this study, our measurements of RI indicate that snapper may not be overtly susceptible to predation upon release, at least when no barotrauma is present. This is because RI was minimal among snapper released after angling simulations most relevant to authentic recreational angling events (i.e. 0.5 min chasing with up to 1 min air exposure), and it is believed that the vigorous condition of these fish would not make them easy targets for predators. Importantly, reflexes that might be associated with reduced predator avoidance, such as the startle response and righting response, remained intact.”

Snapper captured by trawl may be at risk from increased air exposure: 

“It is likely that mortality increases significantly at some point beyond 3 min air exposure in P. auratus but this may not be relevant to recreational angling. Longer periods of air exposure, however, may be present in commercial trawl fisheries where large catches are sorted on deck so knowledge of air exposure tolerance beyond that observed in this study would be useful in this context. Therefore, the existence of a predictive relationship between RI and mortality in snapper remains a possibility but probably requires the inclusion of more extreme air exposure treatment to be clarified in future trials.”

Clearly any capture of snapper that produces barotrauma can be a source of mortality and requires further study in deep water commercial and recreational fisheries.

Measuring reflex action impairment in sole and plaice; preliminary steps to making RAMP

Collection of fish with beam trawl, Jochen Depestele

The beginning steps for measuring reflex action impairment and making a RAMP are detailed in “Calibration tests for identifying reflex action mortality predictor reflexes for sole (Solea solea) and plaice (Pleuronectes platessa): preliminary results” authored by Depestele, J. et al. 2014. Experiments were designed to collect sole and plaice using short hauls of a beam trawl, to test their reflex actions, and to identify consistent reflexes for making RAMP. These experiments followed steps for making RAMP.

A short video demonstrates testing plaice for reflex actions including righting, eye roll (vestibular-ocular response), evade, operculum, mouth, and tail grab. Fish are shown in a series of increasing impairment.

Conclusions from the study included:

Preliminary investigations have been undertaken on-board the RV Belgica to assess the potential presence of a range of reflexes in sole and plaice. A wide range of potential reflexes was investigated prior and during the sea trial, leading to a final selection of seven reflexes with a good potential of being consistently present in fish in a favorably vital condition. Fish in a “perfect” condition could not be retrieved, but 22 individuals of plaice and sole were selected from short hauls and their survival potential was evaluated during 70 hours in on-board holding facilities. Only one sole died, and indicated hence that the control fish for the calibration test serve purpose.

Holding tanks for fish on board RV Belgica, Jochen Depestele

The final selected reflex actions were very similar for sole and plaice, except for one. Forced opening of sole’s operculum did not reveal much resistance of the fish, while holding plaice by its head did not induce curling of the fish. The most consistent reflex actions for sole were called “stabilize, mouth, and tail grab”, followed by the “vestibular-ocular response”. Vital individuals seemingly dig into the sand or stabilize themselves onto the floor of the water-filled box. They also keep their mouth closed when trying to open it with a probe. When fish have stabilized, they respond clearly to grabbing their tail or even tickling it. The “head” reflex was easy to assess, though not always present. However, it is clear that vital soles curled around one’s hand when they had been in holding tanks. This was not that obvious for fish that were just released from the codend. Natural righting was observed regularly, although some individuals remained at their backs for >5 sec and did not return to their natural position at all or only after stimulating them. The consistency could thus be questioned, but good candidate reflexes were proposed for sole, and should be further evaluated. The most consistent reflexes of plaice were the turning of the eyes when the fish was turned around longitudinally. The resistance of plaice to forced opening of the operculum was a clear reaction as well. Not fully consistent, but nevertheless a good indication of the reflexes was the “evade” response and the “tail grab”. When the tail is touched or grabbed in a “good” way (which might require some practice), then the fish swim away, or at least the fins stimulate propulsion. The mouth of plaice was easily opened, but mostly the individuals tried to close it or seemingly opposed to the forced movement.

Our investigations confirmed that on-board holding facilities result in high survival of plaice and sole from very short hauls (<20min). Investigated individuals were non-randomly selected and thus it was not surprising that their physical injuries were limited. These individuals were suitable for developing the reflexes, although they were limited in number (22 for plaice and 22 for sole) and they also did not range over a wide variety of fish conditions (e.g. limited length variability). The seven reflexes from these preliminary investigations are therefore proposed as candidates for the development of a RAMP score for sole and plaice.

The tests of the reflexes were run directly after releasing fish from the codend. When examining the survival from fish that were accommodated for some time (e.g. 48 hours), we noted that they reacted more strongly and had much clearer responses to the reflex tests. In particular the tail grab worked very nicely for sole when their status (alive or dead) was tested. Therefore we suggest that the proposed reflexes are tested once more on surviving individuals of short hauls after an accommodation period of >24hours. Consistency of the outcome of the reflex tests is expected to be improved when the impairment from the catching process is accounting for. Other recommendations for follow-up tests relate to the registration of potential environmental and biological confounding factors.

Survival of the fittest

Bob van Marlen, IMARES

Work is in progress at ICES to define methods for estimating discard survival in reference to the new European Commission Common Fisheries Policy. The ICES newsletter states, “Under the recently reformed European Commission Common Fisheries Policy (CFP), the practice of discarding fish will be phased out, replaced instead with landing obligations. Under the landing obligation, all catches of regulated species must be landed and counted against quotas unless it has been scientifically proven that the species can survive the discarding process. Species that display a high level of discard survivability will be awarded an exemption, meaning that fishers can return these fish to the sea. Unregulated and protected species will continue to be released.”

The ICES WKMEDS focuses on developing guidelines and identifying best practice for undertaking experiments to investigate the survival of organisms discarded from the catches of commercial fisheries. 

Importantly ICES states, “Techniques for estimating survival under review include captive observation, vitality assessment, and tagging and biotelemetry, each with its own advantages and disadvantages. By using a combination of techniques, as WKMEDS suggests, clear synergies can be achieved and challenges overcome.”

"It's an exciting time", state the workshop Chairs Michael Breen and Tom Catchpole, "this group will be central to an international community that are working together to address the important issue of discard survival."

The WKMEDS group is producing a synthesis of previous discard survival and mortality research with the goal to develop an integrated approach for estimating discard survival. The integrated approach is designed to guide scientists and managers in their evaluation of potential candidate fisheries and species for discarding exceptions to the landing obligation of the CFP. The integrated approach incorporates information about the role of fishing conditions as stressors, species sensitivity to stressors, vitality impairment of captured species, estimated discard survival rates, and cost-benefit analysis of methods for survival assessment in fisheries as well as impacts of discarding on fisheries stocks.

Quest for a discard survival predictive scoring system to use on board fishing vessels

Releasing tagged Atlantic cod, John Clarke Russ

The European Union Common Fisheries Policy (CFP) ban on discarding allows for animals to be discarded if “scientific evidence demonstrates high survival rates”. Estimating discard survival for fisheries has become a priority for implementation of the CFP. Limited data on discard survival and mortality is available and methods for estimation have not been standardized. Ideally, a standardized numerical scoring system can be developed and validated, based on readily observable responses and symptoms present in animals that are candidates for discarding and survival. 

RAMP is an example of a predictive scoring system for vitality, survival, and mortality, based on animal reflex actions, barotrauma symptoms, and injury that can be observed in fishing operations where real time decisions must be made about potential discarding. See post for RAMP development and validation; also Davis (2010) and Stoner (2012) for reviews of RAMP method. Other uses for RAMP are in live fisheries, aquaculture, and pollution research and monitoring.

For inspiration and alternative perspectives, examples of validated mortality predictive scoring systems can be found in human and veterinarian intensive care unit (ICU) settings, where patients present with symptoms and disease likely to result in morbidity and mortality (Rockar et al. 1994, Bouch and Thompson 2008, Timmers et al. 2011). Measurements of blood plasma and urine variables commonly made in ICU settings are not contemplated for RAMP since they are not readily made on board fishing vessels.

Below is an example of human ICU mortality prediction using the SAPS II scoring system. Note the similarity to RAMP curves for mortality prediction.

SAPS II mortality predictive scoring system, ClinCalc

Celinski and Jonas (2004) discussed scoring systems developed for the human ICU environment:

“How are scoring systems developed? All available data types and variables can potentially be used to create a scoring system. However, to make it useful, variables have to be selected to be appropriate for the predictive properties of the scoring system. The information must be unambiguous, mutually exclusive, reliable and easy to determine and collect. Ideally, the variables should be frequently recorded or measured.

The variables can be selected using clinical judgement and recognized physiological associations or by using computerized searching of data (collected from patient databases) and relating it to outcome. The variables are then assigned a weighting in relation to their importance in the predictive power of the scoring system (again by using clinical relevance or computerized databases).

Logistic regression analysis, a multivariate statistical procedure, is then used to convert a score to a predicted probability of the outcome measured (usually morbidity or mortality) against a large database of comparable patients. Lastly, the scoring system must be validated on a population of patients independent from those used to develop the scoring system.”

For discard survival prediction, groups of animals, rather than individuals, are the appropriate unit for consideration since proportion mortality is the determined outcome during index development and validation. These groups can represent various scales of resolution in fisheries of interest, i.e., single tows or traps, sets of longline, trap, or gill-net, daily catch.

Jean-Roger Le Gall (2005) discussed the appropriate use of ICU severity scoring systems:

“A good severity system provides an accurate estimate of the number of patients predicted to die among a group of similar patients; however, it does not provide a prediction of which particular patients will in fact die. Using a well-calibrated severity model, we can reasonably expect that approx. 75% of patients with a probability of mortality of 0.75 will die, but we cannot know in advance which of those patients will be among the 25% who will live. Furthermore, these 25% will not have falsified the odds but will have confirmed the validity of the probabilities. 

       The possibility that clinical decisions can be augmented by having an objective (although not always more accurate) assessment of a patient’s severity of illness is appealing. Physicians are interested in severity systems for individual patients as an adjunct to their informed but subjective opinion. Using these tools as part of the decision-making process is reasonable and prudent. Using these tools to dictate individual patient decisions is not appropriate. Decisions will and should remain the responsibility of the individual physician and should be based on a number of criteria, one of which is severity as estimated by a well calibrated scoring system.”

Stacy et al. (2013) discussed development and appropriate use of a predictive scoring system for survival in Kemp’s ridley sea turtles:

“Three mortality prediction indices (MPI) scoring systems were developed using different combinations of blood analytes, with anticipation that at least one of the three would be more accurate in predicting mortality in sea turtles within 7 days after admission. Turtles with higher scores were categorized as physiologically deranged to a degree that could result in death, and turtles that received lower scores were categorized as physiologically stable and likely to survive. Categorization of each turtle was then compared to the known outcome for that individual.

Receiver operating characteristic (ROC) analysis was used to assess the diagnostic performance of each MPI scoring system (Greiner et al., 2000; Giguere et al., 2003). The ROC analysis produces a plot that is used to estimate the area under a ROC curve, which is a summary statistic of diagnostic accuracy. A perfect test [i.e., sensitivity (SE) = 100% and specificity (SP) = 100%] will produce an area under the curve (AUC) = 1. The AUC can be used to distinguish a non-informative test (AUC = 0.5), a less accurate (0.5 < AUC ≤ 0.7), moderately accurate (0.7 < AUC ≤ 0.9), highly accurate (0.9 < AUC < 1), and perfect test (AUC = 1)."

ROC analysis of Kemp's ridley sea turtle mortality predictive scoring system, Stacy et al. 2013

"It is clear that results of mortality prediction indices (MPI) scoring systems cannot be used indiscriminately to make euthanasia decisions, because this would result in euthanasia of some turtles with a falsely positive MPI score that would otherwise survive. As with other health scoring systems in human and veterinary medicine, the MPI scores should not prevent clinicians from providing care to an individual, and euthanasia decisions should only be made in light of numerous other clinical factors, including neurological status, vision, ability to forage, ability to swim, pain and suffering, and duration of illness. Finally, MPI scores may be useful when applied retrospectively in a stranding event for comparison of various treatment outcomes within a facility or among different facilities. Thus, the MPI could provide an objective assessment tool of treatment success and contribute to the advancement of medical care in sea turtles.”

Effect of hypoxia, injury, and facilitated recovery on reflex impairment (RAMP) in migrating sockeye salmon

Sockeye salmon and Dolly Varden, J Armstrong/UW

A study on the effects of hypoxia and injury associated with gill net fisheries and facilitated recovery in sockeye salmon migrating in the Fraser River showed important results for the use of RAMP to measure sublethal effects of stressors (Nguyen et al. 2014).

The authors state:

“Here, we examined sources of delayed fisheries-related mortality in relation to three known factors influencing postrelease behavior and mortality in fish: physiological exhaustion (stress through air exposure), physical damage (via gill net entanglement), and facilitated recovery (using Fraser boxes). We used sockeye salmon (Oncorhynchus nerka) in the lower Fraser River as a model for this research, given conservation concerns regarding a number of sockeye populations (see Cooke et al. 2012). The study was designed to simulate gill net fisheries because high levels of delayed mortality may have important implications for harvest management in exploited and non-target salmon populations. Our primary objective was to distinguish the relative consequences of physical injury and air exposure stress using an experimental approach coupled with reflex assessments (Davis 2010), physiological sampling (non-lethal blood samples; see Cooke et al. 2005), and telemetry tracking of postrelease migration success (Donaldson et al. 2008). Specifically, we used assessments of reflex impairment and blood physiology to characterize the relative impacts of our experimental treatments. Our secondary objective was to test whether Fraser recovery boxes could reduce delayed mortality and improve migration speed for captured fish exposed to varying degrees of stress and injury.”

The study used experimental stressor treatments: C - Captured-only; A - captured and Air exposed; I - captured and Injured; IA - captured and Injured + Air exposed.

The authors found:

“RAMP is intended to be a rapid, simple, and inexpensive means of assessing fish vitality (Davis 2010). It has also been validated as a predictive measure for delayed mortality in coho salmon caught in beach seine fisheries (Raby et al. 2012). RAMP scores indicated sublethal effects resulting from the A treatment but not from the I treatment. Thus, either RAMP may not capture sublethal effects from injuries, even though fish were clearly stressed (elevated plasma lactate and cortisol), or the I treatment used here was not severe enough to impair reflexes. Further research investigating a large range of physical injury might be useful in resolving this issue. Until this is done, we believe it is unwise to rely solely on a RAMP score for predicting delayed mortality of injured migrating adult sockeye salmon. Previous studies show that RAMP scores are positively correlated with intensity of capture stressors (e.g., Davis 2005, 2007; Davis and Ottmar 2006; Humborstad et al. 2009; Raby et al. 2012), but none considered the potential linkage between RAMP and physical injury. Nonetheless, wounds inflicted in fish during capture, which can be highly variable, are a major source of mortality for discards and escapees (Trumble et al. 2000; Suuronen et al. 2005). In the interim, quantitative indexes for physical injuries in fishes have been developed and used in field settings such as visual assessments (e.g., Trumble et al. 2000; Davis 2005; Baker and Schindler 2009) or use of forensic techniques (e.g., fluorescein) to detect nonmacroscopic injuries (Noga and Udomkusonsri 2002; Davis and Ottmar 2006; Colotelo et al. 2009) and might be useful to include when predicting mortality.”

Clearly, further research and validation is needed to establish relationships between RAMP, injury, vitality, and mortality.  As suggested for fish that show barotrauma symptoms, it may be appropriate to consider the inclusion of scoring for presence or absence of injury types in combination with reflex impairment. The effects of injury on reflex impairment differ among species, as shown for fish (Davis and Ottmar, 2006) and crabs (Stoner et al. 2008). Also, at lower levels of stress in some species, reflex impairment may not occur, indicating that the animals are responding to stress in an adaptive manner. Consideration and inclusion of injury in RAMP is important because of it's potential relationship with delayed onset of disease associated with tissue exposure to pathogens.

Approaches for modeling and predicting bycatch mortality

Modeling and prediction of bycatch mortality can be approached in several ways. Efforts can be focused on prediction from knowledge of controlling environmental factors and fishing processes encountered by animals. Alternatively, efforts can be focused on prediction from knowledge of animal condition that integrates effects of fishing factors. A third hybrid approach combines information about animal condition and controlling fishing factors.

Fishing is conducted in freshwater and seawater, with catch retained, discarded, released, or escaped from commercial, recreational, catch and release, and subsistence fisheries. In all cases, knowledge of target and non-target fishing mortality is essential for management and conservation of fisheries stocks and ecosystems. Fishing occurs under a variety of environmental and operational conditions. Examples of fishing gears include trawls, seines, traps, dredges, hook and line, gill nets, lift nets, and falling gear. While immediate mortality is evident for non-target bycatch discards, delayed mortality is generally hidden from view for discards and escapees from fishing gears and operations.

Fishing factors include a range of types for discards and escapees. Master controlling variables include temperature, air exposure, gear injury, fatigue and exhaustion, fish size, barotrauma, and predators. Synergistic effects of combinations of factors can be significant controllers of mortality.

Davis 2002 capture and discard

Suuronen 2005 capture and escape

Bycatch mortality can be modeled by experimental determination of relationships among environmental and operational factors and mortality rates of various species, either under laboratory or field conditions. Since there are an almost infinite number of factor combinations in a fishery, it is important to prioritize the stress and mortality effects of factors and factor combinations. Primary effects are then modeled for mortality rates. 

Effects of fish size, fishing gear type, and temperature on sablefish mortality, AFSC

A second approach to modeling bycatch mortality is to shift focus from environmental and operational fishing conditions to a more limited set of predictors for mortality based on animal condition. These include wounding, physiological impairment, and reflex impairment. Reflex impairment and RAMP have been found to be the most efficient and inclusive predictors of immediate and delayed mortality.

AFSC

RAMP curves for the relationships between reflex impairment and species mortality are constructed under simulated or actual fishing conditions that are expected.

AFSC

Using constructed RAMP curves, bycatch mortality rates can be measured and predicted in fishing operations through time and space by sampling fish from fisheries.

Georges Bank scallop fishery bycatch

Hybrid combinations of the two modeling approaches can be used if data are available. Animals that are captured or escape from fishing gears can be sampled for RAMP while environmental and operational conditions are noted. Then relationships among these factors can be modeled.

Effect of air exposure on Atlantic cod reflex impairment and mortality, Humborstad et al. 2009

RAMP and bonefish recovery from capture stress

Bonefish are typically captured in a high value capture and release sport fishery.  Released bonefish can be subjected to high rates of predation if predators are present and stress levels are high.  Recovery from capture stress is important for survival and maintenance of bonefish fishery stocks.  Recovery technology including holding bonefish in bags or live wells prior to release has been evaluated by Brownscombe et al. 2013.  They used measurements of reflex impairment (RAMP), locomotory activity, and predation to test recovery from capture stress and survival after release.

Bonefish Key West Flats Fishing

Practical application of RAMP was suggested by Brownscombe et al. 2013. "Bonefish anglers may be able to use RAMP to assess bonefish condition, and make educated decisions on whether to release the fish, or retain it for a short period to facilitate recovery. Likewise, if water temperatures and bonefish impairment scores are very high, responsible anglers can recess until conditions are more favorable."

Here are some excerpts from their paper regarding the validation of RAMP to measure capture stress and predict delayed mortality after release:

"The primary objective of this study was to evaluate the effectiveness of retaining bonefish in recovery bags for reducing short-term locomotory impairment when subjected to angling-related stressors, and whether potential improvements in swimming ability translated to increased survival.

Reflex indicators have recently been deemed effective predictors of mortality (Davis, 2010; Raby et al., 2012), and could be used by anglers to evaluate in which instances fish would benefit from recovery. We predicted that fish retained in recovery bags would exhibit lower reflex impairment, as well as higher locomotory ability and survival than those immediately released.

We validated the use of reflex action mortality predictors (RAMP) (Davis, 2005, 2010) to assess bonefish vitality after 0, 2, 4 and 6 min of air exposure. The 0-minute assessments (n = 30) occurred prior to air exposure on fish from all treatments, while bonefish in the 2-minute treatment (n = 20) were those used in recovery bag experiments (see below), and 4, 6 min treatments (n = 5) were conducted on alternate fish. Five predictors were measured; tail grab, equilibrium (orientation), body flex, head complex, and vestibular-ocular response (VOR). These predictors were chosen because Raby et al. (2012) found that they were strong predictors of coho salmon (Oncorhynchus kisutch) mortality after being caught in commercial nets, and all these predictors can be easily and quickly measured by bonefish anglers. RAMP was assessed in the same manner by Raby et al. (2012). The presence of a tail grab response was assessed by grabbing the fish's tail while it is submerged in water; it was considered impaired if the fish did not attempt to swim away from the handler. Equilibrium was assessed by rolling the fish upside down in water; impairment was indicated when the fish was unable to right itself within 3 s. Body flex was tested by holding the fish by the middle of the body in air; it was considered impaired if the fish made no attempt to struggle free. Head complex was considered impaired if while holding fish in air, a regular pattern of ventilation of the fish's operculum was not observed for at least 5 s. VOR was assessed by rolling the fish back and forth in air; it was considered impaired if its eyes did not roll to maintain the same pitch and track the angler. Higher RAMP scores indicated greater impairment.

Based on the responsiveness of bonefish to the RAMP indices, we used them to evaluate the utility of the recovery bags. After air exposure, bonefish released with accelerometers had similar RAMP scores between immediate release (2.8 ± 0.14) and recovery (2.7 ± 0.14) treatments. However, after retention in a recovery bag for 15 min, all bonefish had RAMP scores of zero (i.e., full recovery).

Our results demonstrate that retaining bonefish in recovery bags for 15 min reduced locomotory impairment upon release during the critical time period where most predation occurs, and this practice has the potential to increase survival after catch-and-release angling. Presumably, retaining bonefish in a live well with ambient oxygen levels (Shultz et al., 2011) would have a similar benefit if an angler had access to a boat.

The five impairment indicators we tested on bonefish provided a gradient in impairment scores that related to the degree of stressor (i.e., 0–6 min of air exposure). RAMP scores have been correlated with stressor duration and mortality for a number of fish species (Davis, 2005, 2007; Davis and Ottmar, 2006; Humborstad et al., 2009; Raby et al., 2012). Indeed, the duration of a stressful event increases the level of physiological disturbance in bonefish (Suski et al., 2007; Donaldson et al., 2008), while longer handling times and air exposure durations result in higher post-release predation rates (Danylchuk et al., 2007a). In this study, bonefish that were equipped with accelerometers exhibited moderate impairment scores after 2 min of air exposure, while no impairment was detected after 15 min of retention in a recovery bag, and fish from the recovery treatment exhibited significantly higher levels of activity upon release. Therefore RAMP scores appear to be a good indication of bonefish vitality. 

The impairment indicators tail grab, equilibrium, and body flex were the first to become impaired in bonefish, and impairment levels within these predictors did not vary with increased stress duration. This was likely because bonefish were all highly impaired at the lowest level of stress we inflicted. Indeed, a previous study found roughly that 50% of bonefish lose equilibrium after angling events (Danylchuk et al., 2007a), while 95% of bonefish lost equilibrium after simulated angling stress (2 min of air exposure) in this study. These three predictors may provide an indication of impairment levels with lesser degrees of stress. Head complex was the next to become impaired at 4 min of air exposure, followed by VOR at 6 min. Therefore head complex and VOR predictors are indicative of very high levels of physiological disturbance in bonefish. This predictor-specific pattern of impairment in bonefish is nearly identical to that of coho salmon (see Raby et al., 2012)."