Reflex impairment in largemouth bass shows interactions of gear type, fight time, and air exposure

Largemouth bass, Bemep/Flicker

Cooke et al. 2016 examined reflex impairment in largemouth bass captured during the summer (25-27^oC). Excerpts from their paper detail study findings: 

“…little is known about how gear strength and fight time interact with air exposure duration to ultimately influence the level of exhaustion experienced by fish at time of release. Here we systematically varied fishing gear strength (ultralight versus medium-heavy) and air exposure duration (0 versus 120 s) when targeting Largemouth Bass Micropterus salmoides. We relied on reflex impairment (using five different reflexes) as a real-time indicator of fish condition.”

“One of the more interesting observations from this study was that fish that were landed rapidly and thus in better condition were more difficult to handle, which led to longer air exposure. We are aware of anglers and scientists that have mused about the trade-offs between fight time and ease of handling, but to our knowledge this is the first study to formally assess this idea.”

“In this study we used two extremes in gear type and suggest that moderate strength gear likely represents the best compromise in terms of achieving an appropriate level of exhaustion that would facilitate handling and hook removal without leading to complete exhaustion.”

Reflex impairment in captured largemouth bass, Cooke et al. 2016.

“Using reflex indicators, we showed clearly that there was a gradient in reflex impairment with Largemouth Bass; fish captured on UL gear had significantly higher reflex impairment than those captured with MH gear with no air exposure, while fish captured with both gear types had similarly high reflex impairment when exposed to the air.”

A bigger picture: factors and traits that contribute to vitality and survival of discards in fisheries

ICES WKMEDS4 report (2015)

A conceptual model for discard survival in fisheries is developed in the ICES WKMEDS4 report (2015). In this concept, survival is linked to species sensitivity, injury, and predation, through fishing factors, environment, and size. The expanded view shows potential factors and traits in more detail.

ICES WKMEDS4 report (2015) Click on images.

Triage for captured and released fisheries species: research and survival

Will they survive? (The Guardian, 2013)

Vitality impairment can be linked to post-capture mortality in fisheries bycatch. Vitality impairment can be estimated by direct observation of animal activity, responsiveness, and injury. For each critical fisheries species in crabs, fishes, sharks, and turtles, reflex actions that are consistently present in healthy, uninjured individuals are listed as control levels. Impairment is signified by loss of reflex action types and addition of injury types after capture.  

Reflex actions are fixed, consistent animal behavior patterns that can be triggered by perception of external stimuli (light, sound, smell, gravity, touch). Stimulation of reflex actions is not controlled by body size, motivation, strength of stimulus, or fear. Reflex action traits summed as a whole animal can be an expression of vitality (Davis 2010). In contrast, volitional behavior can be altered by body size, motivation, strength of stimulus, fear, cognition, and as such is not a controlled measure of vitality.

With the species reflexes and potential injury lists, observations of captured animals can be made in commercial and sport fisheries. Patterns of significant impairment can be determined and related to fishing context and species (Raby et al. 2015). These patterns help identify the relative effects of fishing gears, handling, and physical factors (air, temperature, light, pressure) on impairment and potential survival and mortality.

Figure shows overlap between information on animal physiology and fisheries biology, adapted from Horodysky et al. 2015 and modified to show vitality information. Measures of vitality include reflex impairment and injury, which are whole animal measures that are ecologically relevant, linking physiological and population level research and hypothesis testing. Volitional behavior is coordinated whole animal movements beginning with perception and motivation, followed by attraction and aversion to various stimuli (injury, threat, food, shelter, species mates, migration).

Patterns of vitality impairment can guide research questions and priorities to triage fisheries for treatment of bycatch mortality and enhancement of survival. Vitality impairment can measure the efficacy of engineering fishing gears to increase bycatch survival. 

Observing vitality impairment

Animal vitality can be measured by observing species traits associated with activity, responsiveness, and injury. For each species, a group of reflex actions can be observed that are consistently present in healthy animals. As vitality becomes impaired, reflex action traits disappear and injury traits may begin to appear. 

Activity, responsiveness, and injury for measurement of vitality impairment (Benoît et al. 2010). 

Fisheries show gradients of stressors associated with capture, handling, and release or escape. Discard mortality, survival, and vitality impairment are controlled by stressor gradients.

Gradients of mortality and simulated stressors in sablefish fisheries; water temperature and gear type including trawl (time), longline, pot. Smaller fish are more sensitive to stressors (AFSC).

Vitality impairment gradients are associated with stressors and can be used to predict survival and delayed mortality for populations of impaired animals. Vitality impairment gradients can be measured by identifying classes of health condition; excellent, good, poor, and moribund based on rapid observation and impression of animal injury and activity (Benoît et al. 2015). 

The resolution for observations of vitality impairment gradients can be increased by including more information. RAMP is an example of this approach (Davis and Ottmar 2006, Davis 2010). A list can be made of reflex actions present in control animals and possible injuries. Then presence or absence of listed traits is observed after exposure to stressors. Increasing impairment is associated with stress effects and morbidity.

Reflex actions observed in snapper by McArley and Herbert 2014.

Relationships between vitality impairment and survival or delayed mortality can be experimentally determined. Then predictions of stress effects in other settings with similar stressors can be made by measuring vitality impairment associated with stressors, without the need to hold or tag animals. Vitality impairment can be rapidly observed in sampled populations as an additional factor to evaluate stressor effects and is a useful indicator of animal health and stress status, that can be validated experimentally.

Reflex impairment and mortality for individuals (A) and groups (B) of Atlantic cod with 95% confidence intervals (Humborstad et al. 2009). 

Snow crab discard mortality

Snow crab in Bering Sea pot fishery (ASMI).

Over 19,000 snow crab were evaluated in Bering Sea pot fisheries 2010-2012 for impairment using the RAMP method (Urban 2015). The estimated discard mortality rate was 4.5% (s.d. = 0.812), significantly below the rate used in stock assessment models. The author concludes: “ In this study, the results of RAMP observations showed that at the range of winter temperatures typically encountered by the Bering Sea snow crab fishery, nearly all discarded crab experienced no reflex impairments. Therefore, we estimate that they should have only a 4.8% chance of short-term mortality. Injuries caused by the fishery occurred at very low levels and so should also have a minimal effect on discard mortality rates. However, because long-term survival rates and the effects of reduced crab vitality are difficult to predict, an estimate of the total impact of discard practices on snow crab stocks is not possible. Even with these uncertainties, the current empirical evidence indicates that the assumed discard mortality rate of 50% is conservative.”

Figure 1. The upper panel shows the relationship between the temperature at the snow crab sorting table and the predicted mortality of snow crab based on reflex impairments. Error bars indicate the 95% CI. The lower panel shows the proportions of the temperatures recorded, while the observations were being made during the 2010–2012 fisheries (Urban 2015).

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.

Belly up: Righting reflex action time to recovery correlated with delayed mortality?

Upside down fish in market tank (Hong Kong)

RAMP incorporates presence/absence of several reflex actions and injuries to measure vitality impairment and potential delayed mortality. A simpler method may be possible by measuring time for recovery of orientation when fish are placed upside down in water. This method can be tested.

Place a fish upside down in water and observe the time until the fish returns to normal orientation. This duration is a measure of vitality impairment. Longer recovery times indicate greater vitality impairment and data can be included in statistical models for relationships among fishery stressors, injury, righting time, and delayed mortality. We can test the relationship between righting impairment and delayed mortality. 

Righting reflex action is a central behavior that is the nexus of neural, muscle, and organ actions and is intimately linked with loss of physiological regulation associated with stressor exposure.  Olfactory impairment is another example of a central function that is correlated with delayed mortality (in humans, Pinto 2014).

Body orientation is a sensitive measure of fish consciousness. Presence or absence of righting can be included in the RAMP score. Loss and recovery of orientation is a well known symptom for induction of and recovery from fish anesthesia and is used as an indicator of morbidity and delayed mortality in stress experiments (Davis and Ottmar 2006, Szekeres et al. 2014, Raby et al. 2015).  

Measuring replicate animals for the time to righting recovery and delayed mortality after a stressor experiment can test the correlation between righting impairment and delayed mortality. If the correlation between righting and delayed mortality is valid and strong, then we have a rapid method for predicting discard mortality on board fishing vessels without need for holding or tagging fish to confirm their survival. Research groups on fishing vessels can observe fish during catching, landing, sorting, and discarding under differing stressors; seasons, water temperatures, tow durations, catch quantities, species mixes, and sorting times.

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.

Fisheries studies that have used and documented reflex actions or injury for vitality impairment testing

The table presents selected examples of reflex action & injury vitality impairment testing.

See also a list of reflex actions and injury that may be scored for vitality impairment.

Citations:

AFSC, Alaska Fisheries Science Center 2014 Observer Sampling Manual.

Fisheries Monitoring and Analysis Division, North Pacific Groundfish Observer Program. Seattle, WA.

Barkley, A.S. & Cadrin, S.X. 2012. Discard mortality estimation of yellowtail flounder using reflex action mortality predictors. Trans. Am. Fish. Soc. 141:638-644.

Benoît, H.P., Hurlbut, T., and Chassé, J. 2010. Assessing the factors influencing discard mortality of demersal fishes using a semi-quantitative indicator of survival potential. Fish. Res. 106:436-447.

Braccini, M., Van Rijn, J., and Frick, L. 2012. High post-capture survival for sharks, rays and chimaeras discarded in the main shark fishery of Australia? PLoS ONE 7: e32547.

Brownscombe, J.W., Thiem, J.D., Hatry, C., Cull, F. Haak, C.R., Danylchuk, A.J., and Cooke, S.J. 2013. Recovery bags reduce post-release impairments in locomotory activity and behavior of bonefish (Albula spp.) following exposure to angling-related stressors. J. Expt. Mar. Biol. Ecol. 440:207-215.

Brownscombe, J.W., Nowell, L., Samson, E., Danylchuk, A.J., & Cooke, S.J. 2014. Fishing-related stressors inhibit refuge-seeking behavior in released subadult Great barracuda. Trans. Am. Fish. Soc. 143:613-617.

Campbell, M.D., Patino, R., Tolan, J., Strauss, R., and Diamond, S.L. 2010.

Sub-lethal effects of catch-and-release fishing: measuring capture stress, fish impairment, and predation risk using a condition index. ICES J. Mar. Sci. 67:513-521. 

Campbell, M.D., Tolan, J., Strauss, R., and Diamond, S.L. 2010. Relating angling-dependent fish impairment to immediate release mortality of red snapper (Lutjanus campechanus). Fish. Res. 106:64-70.

Danylchuk, A.J., Suski, C.D., Mandelman, J.W., Murchie, J.W., Haak, Brooks, A.M.L., and Cooke, S.J. 2014. Hooking injury, physiological status and short-term mortality of juvenile lemon sharks (Negaprion bevirostris) following catch-and-release recreational angling. Cons. Physiol. 2:cot036.

Davis, M. W. 2007. Simulated fishing experiments for predicting delayed mortality rates using reflex impairment in restrained fish. ICES J. Mar. Sci. 64:1535-1542.

Davis, M.W. & Ottmar, M.L. 2006. Wounding and reflex impairment may be predictors for mortality in discarded or escaped fish. Fish. Res. 82:1-6.

Depestele, J., Buyvoets, E., Calebout, P., Desender, M., Goossens, J., Lagast, E., Vuylsteke, D., and Vanden Berghe, C. 2014. Calibration tests for estimating reflex action mortality predictor for sole (Solea solea) and plaice (Pleuronectes platessa): preliminary results. ILVO-communication. Report nr. 158. 30p. 

Diamond, S.L. & Campbell, M.D. 2009. Linking “sink or swim” indicators to delayed mortality in red snapper by using a condition index. Mar. Coast. Fish.: Dynamics, Manag. Eco. Sci. 1:107-120.

Donaldson, M.R., Hinch, S.G., Raby, G.D., Patterson, D.A., Farrell, A.P., and Cooke, S.J. 2012. Population-specific consequences of fisheries-related stressors on adult sockeye salmon. Physiol. Biochem. Zool. 85:729-739.

Hammond, C.F., Conquest, L.L., and Rose, C.S. 2013. Using reflex action mortality predictors (RAMP) to evaluate if trawl gear modifications reduce the unobserved mortality of Tanner crab (Chionoecetes bairdi) and snow crab (C. opilio). ICES J. Mar. Sci. 70:1308-1318.

Hannah, R.W. and Matteson, K.M. 2007. Behavior of nine species of Pacific rockfish after hook-and-line capture, recompression, and release. Trans. Amer. Fish. Soc. 136:24-33.

Humborstad, O-B., Davis, M.W., and Løkkeborg, S. 2009. Reflex impairment as a measure of vitality and survival potential of Atlantic cod (Gadus morhua). Fish. Bull. 107:395-402. 

McArley, T.J. & Herbert, N.A. 2014. Mortality, physiological stress and reflex impairment in sub-legal Pagrua auratus exposed to simulated angling.  J. Expt. Mar. Biol. Ecol. 461:61-72.

Raby, G.D., Donaldson, M.R., Hinch, S.G., Patterson, D.A., Lotto, A.G., Robichaud, D., English, K.K., Willmore, W.G., Farrell, A.P., Davis, M.W., and Cooke, S.J. 2012. Validation of reflex indicators for measuring vitality and predicting the delayed mortality of wild coho salmon bycatch released from fishing gears. J. Appl. Ecol. 49:90-98.

Stoner, A.W. 2012. Assessing stress and predicting mortality in economically significant crustaceans. Rev. Fish. Sci. 20:111-135.

Stoner, A.W., Rose, C.S., Munk, J.E., Hammond, C.F., and Davis, M.W. 2008. An assessment of discard mortality for two Alaskan crab species, Tanner crab (Chionoecetes bairdi) and snow crab (C. opilio), based on reflex impairment. Fish. Bull. 106:337-347.

Szekeres, P., Brownscombe, J.W., Cull, F., Danylchuk, A.J., Shultz, A.D., Suski, C.D., Murchie, K.J., and Cooke, S.J. 2014. Physiological and behavioural consequences of cold shock on bonefish (Albula vulpes) in The Bahamas. J. Expt. Mar. Biol. Ecol. 459:1-7.

Trumble, R.J., Kaimmer, S.M., and Williams, G.H. 2000. Estimation of discard mortality rates for Pacific halibut bycatch in groundfish longline fisheries. N. Amer. J. Fish. Manag. 20:931-939.

Yochum, N., Rose, C.S., and Hammond, C.F. 2015. Evaluating the flexibility of a reflex action mortality predictor to determine bycatch mortality rates: A case study of Tanner crab (Chionoecetes bairdi) bycaught in Alaska bottom trawls. Fish. Res. 161:226-234.

RAMP is a component of an integrated conservation approach to coho salmon bycatch mortality management

Raby et al. 2014

Results of Raby et al. 2014 demonstrate the integration of vitality impairment and coho bycatch mortality estimation and management.

“We have provided an estimate of bycatch mortality for an endangered population of coho salmon captured in an aboriginal beach seine fishery, based on three years of tracking fish released from the fishery.” 

“Among all the variables we tested as predictors of mortality, none were significant except for RAMP score, whereby fish with higher RAMP scores (more impaired) were less likely to be successful migrants (Table 3, Fig. 4).”

Distinguishing between natural mortality and bycatch mortality. 

“An alternate approach to calculating a bycatch mortality rate that attempts to distinguish bycatch from natural mortality, is to use RAMP scores and their mortality rates at each level of impairment, and assume negligible bycatch mortality for the fish that were least impacted (vigorous at release).”

“Since some in-river mortality is natural, there is a need to attempt to differentiate mortality caused by the capture itself. To do so, RAMP scores can be used whereby coho salmon released with little or no reflex impairment (vigorous) are assumed to experience no post-release bycatch mortality. Using that conservative assumption, the post-release mortality rate for those fish can then be used as a baseline within the data set. Additional mortality above that baseline that occurs at higher levels of reflex impairment can then be assigned to the fishery (see Fig. 4).”

Using RAMP to monitor condition of bycatch and improve their survival

“The expanded validation of the RAMP approach in the present study provides confirmation that this simple technique is ready for use in this fishery if needed (Raby et al. 2012). The observers in the fishery could easily be taught how to conduct RAMP assessments to monitor the condition of bycatch in real time, provide advice to their crews on how to improve fish condition, and make decisions about whether individual fish should be revived using recovery bags.”

Human delayed mortality can be predicted using olfactory impairment

Olfactory impairment in humans was measured by error rate in olfaction tests. Increasing number of errors in olfaction tests were related to increasing 5-year mortality rates in a logistic regression (PLoS ONE). 

The human logistic relationship between olfactory impairment and 5-year delayed mortality is a powerful method for predicting delayed mortality and is similar to other animal RAMP relationships between reflex impairment, injury, and delayed mortality. Olfactory impairment can be easily measured in human and animal clinical settings and can easily and automatically be measured in aquaculture contexts by analysis of animal distributions and activity in rearing facilities. Given the fundamental nature of olfaction, one would expect the relationship between olfactory impairment and delayed mortality to be generally present among animal phyla and this can be tested in clinical and field settings.

Pinto et al. 2014 state, “We are the first to show that olfactory dysfunction is a strong predictor of 5-year mortality in a nationally representative sample of older adults. Olfactory dysfunction was an independent risk factor for death, stronger than several common causes of death, such as heart failure, lung disease and cancer, indicating that this evolutionarily ancient special sense may signal a key mechanism that affects human longevity. This effect is large enough to identify those at a higher risk of death even after taking account of other factors, yielding a 2.4 fold increase in the average probability of death among those already at high risk (Figure 3B). Even among those near the median risk, anosmia increases the average probability of death from 0.09 (for normal smellers) to 0.25. Thus, from a clinical point of view, assessment of olfactory function would enhance existing tools and strategies to identify those patients at high risk of mortality.”

(PLoS ONE)

The human study controlled for the mortality effects of age, gender, socioeconomic status, and race. Additionally, “We excluded several possibilities that might have explained these striking results. Adjusting for nutrition had little impact on the relationship between olfactory dysfunction and death. Similarly, accounting for cognition and neurodegenerative disease and frailty also failed to mediate the observed effects. Mental health, smoking, and alcohol abuse also did not explain our findings. Risk factors for olfactory loss (male gender, lower socioeconomic status, BMI) were included in our analyses, and though they replicated prior work [41], did not affect our results.” Note that the study did not control for effects of possible episodic exposure to toxins or injury that may result in temporary or permanent olfactory impairment not related to death.

Olfactory response is an involuntary response to a stimulus, and may be considered a reflex action. In the human study, presence or absence of smell detection for rose, leather, orange, fish, and peppermint were summed and related to delayed mortality. Olfactory responses to various substances can be scored as present or absent and summed to predict delayed mortality. In the same way, the RAMP method is an example of presence-absence scoring with summation of reflex impairment and injury scores to predict delayed mortality.  Measuring and summing whole animal responses, i.e., olfaction, reflex actions, and injury to stimuli is a powerful method for observing the effects of stressors and aging on delayed mortality.   

“We believe olfaction is the canary in the coal mine of human health, not that its decline directly causes death. Olfactory dysfunction is a harbinger of either fundamental mechanisms of aging, environmental exposure, or interactions between the two. Unique among the senses, the olfactory system depends on stem cell turnover, and thus may serve as an indicator of deterioration in age-related regenerative capacity more broadly or as a marker of physiologic repair function [13].”

Clearly, measurement and summation of presence-absence for whole animal involuntary characteristics (olfaction, reflex actions, and injury) is a powerful way to predict delayed mortality in humans and other animals.

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.

Making and Using RAMP in Fisheries

A video is available that explains making and using RAMP in fisheries.

Why is vitality impairment related to mortality?

By definition, healthy animals have full vitality. Vitality becomes impaired as animals become stressed by capture and handling. Severe vitality impairment can result from the effects of physical injury or other stressors, e.g., fatigue, temperature, light, sea state, and air exposure. Maladaptive stress responses or critical injury associated with severe vitality impairment can result in immediate and delayed mortality.

Why score reflex actions and injury?

Reflex actions are fixed behavior patterns that are directly related to vitality impairment, without control by volitional behavior factors, e.g., motivation, hunger, fear, shelter seeking, migration, and reproduction. Reflex actions reflect the state of neural, muscle, and organ functions.

Injuries are directly related to vitality impairment because they can control neural, muscle, and organ functions.

Scoring vitality impairment in general

Any type of reflex action or injury that is related to vitality can be summed to score vitality impairment. The important point is that a sum of presence/ absence scores for vitality characteristics produces an index of vitality impairment. This vitality index can then be used as a measure of variability for sublethal stressor effects in fisheries, as well as a validated indicator and predictor of mortality and survival.

Steps for making and using RAMP in fisheries.