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). 

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.

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.”

Using RAMP to help automate aquaculture operations

Net pen for fish culture (NOAA).

Aquaculture of animals in tanks and net pens requires monitoring and maintenance of vitality, health, and normal behavior for efficient and economic operations. Presently, health is monitored by sampling for disease outbreaks, while vitality and behavior are observed by aquaculture technicians during the course of their daily activities of feeding, cleaning, and operation of facilities.

Inside net pen for fish culture (NOAA).

In tanks and net pens, animals can swim and feed normally. They can also respond to stimuli administered inside their rearing environment, such as light flashes, sound bursts, food scent, and touch. Responses to these stimuli can be in the form of reflex actions such as startle, orientation, depth distribution, aggregation, and dispersal. These reflex responses can be observed remotely and automatically using video, infra-red, and sonar technology in light and dark conditions. Reflex responses can be recorded and summed as RAMP scores for measures of impairment and correlation with mortality.

RAMP can be a quantitive measure of animal state and be used to help automate aquaculture monitoring and maintenance. When impaired responses to stimuli are observed, alarms can be triggered and technical staff can be alerted to changes in animal vitality, health, and behavior. Then on site alteration of operations can bring rearing conditions back to nominal states and return animals and their reflex actions to vitality and health.

Future research in aquaculture can consider the use of RAMP and automated reflex testing for development of efficient operation protocols and quality assurance. RAMP can also be used as a research tool for testing and validating new designs for aquaculture operations that optimize animal vitality and health.

Choices for testing reflex impairment in RAMP

Reflexes can be tested in unrestrained or restrained animals. Tests can be adapted to experimental or operational conditions needed in specific situations. A reflex action is scored not impaired (0) when strong or easily observed and scored impaired (1) when not present, weak, or there is doubt about presence. Reflex impairment scores for an individual animal are then summed and divided by the total observable impairments possible to calculate proportion impairment. Impairment score is then correlated with mortality to produce RAMP. RAMP can be expanded when appropriate to include testing for reflex impairment, barotrauma, and injury.

Below are some examples of previous reflex testing in fish, crustaceans, and turtles. Davis 2010 has shown several types of fish reflexes with impairment after stress induction. In free swimming fish, studied reflexes included orientation where the fish should normally be upright, righting reflex where the fish returns to an upright position and the startle response in which the fish shows rapid forward motion in response to stimuli (Lutnesky and Szyper 1990; Artigas et al. 2005; Davis and Ottmar 2006). In restrained fish, studied reflexes included body flex upon restraint where the fish attempts to escape when restrained, dorsal fin erection in which the fins become erect when the fish is restrained, operculum and mouth closure where the operculum or mouth clamps shut when lifted or opened, the gag response where the fish opens its mouth and flexes the body when the throat is stimulated and the vestibular–ocular response (VOR) shown by eye rolls when the body is rotated around the long axis (Trumble et al. 2000; Davis 2007). Other studies of reflexes in free swimming fish have included atonic immobility, dorsal light reaction, and optomotor and optikinetic responses (Douglas and Hawryshyn 1990; McCormack and McDonnell 1994; Wells et al. 2005; Hasegawa 2006).

Raby et al. 2012 used a reflex impairment index for coho salmon modified from the previously developed RAMP method (Davis 2005, 2007). Immediately prior to release, all tagged and biopsied fish were tested for the presence of five reflexes that were consistently present in control, excellent condition fish. Each reflex was assessed categorically (0 = unimpaired, 1 = impaired) in a conservative matter – that is, if the handler had doubt as to whether the reflex was present, it was recorded as being impaired. Reflexes tested were the following: tail grab, body flex, head complex, vestibular-ocular response (VOR) and orientation. Presence of the tail grab response was assessed by the handler attempting to grab the tail of the fish with the fish submerged in water (in a fish bag or holding trough); a positive response was characterized by the fish attempting to burst-swim immediately upon contact. The body flex response was tested by holding the fish out of water using two hands wrapped around the middle of the body. The fish actively attempting to struggle free was characterized as a positive response. Head complex was noted as positive if, when held out of water, the fish exhibited a regular pattern of ventilation (for ∼5 s) observable by watching the opening and closing of the lower jaw. VOR was observed by turning the fish on its side (i.e. on a lengthwise axis) out of water. Positive VOR was characterized by the fish’s eye rolling to maintain level pitch, tracking the handler. Finally, upon release, each fish was placed upside-down in the river just below the surface: a positive orientation reflex was noted if the fish righted itself within 3 s. The entire reflex assessment took ≤ 20 s to complete and was always conducted on fish upon release. If a fish was too vigorous to allow researcher handling and assessment of reflexes, it was assigned an unimpaired status for all reflexes. The reflex actions included in our protocol are thought to be sufficiently varied that they involve different neurological pathways and/or muscle groups such that there are no redundancies. For example, some of the reflexes are part of the autonomous nervous system (head complex, i.e. respiration), while others clearly are not (tail grab, body flex). Moreover, using this RAMP protocol with Pacific salmon, no two reflexes in the suite of five are consistently present/absent together (see Results; G. Raby, unpublished data). From the reflex results for each fish, we calculated a RAMP score: a simple proportion of the five measured reflexes that were impaired in an individual fish (0 = no reflexes impaired, 1 = all reflexes impaired; Davis 2007).

Humborstad et al. 2009 in Atlantic cod.

Barkley and Cadrin 2012 in yellowtail flounder.

Campbell et al. 2009 in red snapper.  Following exposure to rapid decompression, and removal from the hyperbaric chamber, external symptoms of barotrauma were noted and the reflex responses tested.  The barotrauma – reflex(BtR) score developed was modified from the RAMP procedure developed by Davis and Ottmar (2006). All barotrauma incidences observed and reflex responses tested were categorical in nature (1 = unimpaired state, 0 = impaired state). Fish were examined for a suite of barotraumas, including expanded abdominal cavity (tightened air bladder), stomach everted and protruded from the oral cavity, intestine protrusion out of the anus, exophthalmia (eyes bulging), subcutaneous hemorrhaging, and activity level. Reflex response testing was performed out of the water over 1 min, with the fish restrained so that each test could be done in isolation. Reflex responses tested included gag, opercular, dorsal spine, vestibular-ocular (VO), and tail-flex responses. The gag response was tested by inserting a narrow probe into the oesophagus; a positive response was noted as involuntary-muscle contractions intended to dislodge the probe from the oesophagus. The opercular response was measured by observing whether the fish actively attempted to ventilate the gills (gilling). Active gilling was considered a positive response. The dorsal-spine response was tested by moving a probe over the dorsal spine, causing it to fold back. Positive dorsal spine response was noted when spines returned to an erect defensive position. VO response was observed by turning the fish along its lateral axis. Positive VO was noted when the eye rotated within the orbit and refocused on a fixed position (the observer). Tail-muscle flex was tested by inserting a syringe needle into the hypaxial musculature of the subject. Positive tail-flex response was noted if the muscle contracted. If the subject demonstrated hypaxial-muscle contraction (tail flapping) before insertion of the needle, it was considered positive and no needle insertion took place. To calculate the BtR score, the total number of barotraumas and reflex responses present was summed, divided by the total number possible, and subtracted from 1 [BtR = 1 - (summed individual responses/total responses possible)]. A BtR score close to 0 indicated a fish with low impairment, and a score close to 1 indicated reflex impairment or elevated level of trauma. Following BtR observations, the subject was placed in a test arena for predation simulation and measurement of performance responses. For the thermocline-exposed fish (T), the test arena had previously been heated 7oC above ambient, and for the non-thermocline-exposed fish (NT), it was kept at ambient temperature. Predator attack was simulated from behind a screen by rapidly thrusting a dipnet directly at the lateral surface of the fish. Predator simulations were administered at 0, 5, 10, and 15 min post-decompression, which allowed investigation of the time-course of impairment and estimation of recovery times. Response variables measured were closest simulated predator approach distance (AD, cm), maximum burst swimming speed attained (BSS, cm/s), and the amount of time the fish spent reacting to the stimuli, or the duration of the response (Dur, s). A video camera (Sony DCR-TRV117, 32 frames/s) was mounted above the testing arena to film the sequence of predator attack simulation and subject response. A grid was placed on the bottom of the test arena to assist in calculations of AD and swimming speed. 

Stoner 2012 in crustaceans.

Stoner 2012 in spot prawns.

Stoot et al. 2013 in turtles.

Sorting wild-caught animals for high quality and live market

Many fishers are becoming aware of the value added to catch by controlling and accounting for animal vitality and food quality.  Fish and crabs that are stressed by capture and then transported to markets, either as live animals or freshly killed, have lesser value for discriminating consumers.  Vendors and educators for sea- and fresh-water food are developing programs to account for the methods and sustainability of food capture and handling.

Accounting for animal vitality and quality requires information about capture methods and associated impacts on ecosystem structure and function.  Sorting of catch is important for maintaining high market quality.  Rapid transport of catch to market can insure fresh product.  All of these aspects of fishing can benefit from the information that RAMP supplies.

Changes in the design of fishing gear and fishing methods that improve catch vitality and decrease bycatch can be tested and validated using RAMP to measure animal stress responses to the stressors of capture and handling.

Turtle escaping trawl

Fish or crabs can be captured and then transported to net pens for holding and later release onto markets.  Planned supplies of fresh product stabilize markets and increase the value of the catch.  These methods of capture and marketing require the availability of animals with best vitality and quality to reduce transport and holding costs associated with poor quality catch.  RAMP has been proposed for use in an Atlantic cod capture-based aquaculture system.

Capture-based tuna

Increasing use of combinations of wild fishing and aquaculture rearing requires information on animal quality, vitality, and fitness.  RAMP can supply this information used to test and validate the design and use of new aquatic food supply systems and sources.  Because RAMP is a cheap, easy, and immediate source of critical data on animal vitality and fitness, it can be an important component for efficient economies of operation in aquatic food production.

Capture-based aquaculture

The ornamental fish trade has a big impact on wild fish populations and is in critical need of information on fish vitality and fitness in capture, holding, transport, and marketing aspects of the industry.  RAMP can easily, effectively, and economically supply this needed information to improve the sustainability and ethics of this growing industry.

Ornamental fish trade

Handling, transporting, and processing animals often involves the use of anesthesia. Reflex impairment and RAMP can be used to assess the induction and recovery from anesthesia. 

Induction of anesthesia in fishes is described by loss of activity and responsiveness (Neiffer and Stamper 2009); “With proper dosing, induction with immersion drugs usually occurs within 5 to 10 minutes, but may take longer via other routes. Induction is marked by decreases in caudal fin strokes, swimming, respiratory rate, and reaction to stimuli; the drop in caudal fin stroke activity is usually the first sign, followed by loss of equilibrium and response to stimuli. At surgical anesthesia there is total loss of muscle tone and a further decrease in respiratory rate. A firm squeeze at the base of the tail can be an effective way to determine response to stimuli: if the animal does not respond, general anesthesia has taken effect (Harms 2003; Stetter 2001).”

Figure shows loss and recovery of swimming and equilibrium (behaviours), startle response to probe stimulation (responses), and vestibular-ocular response and rhythmic breathing (reflexes) associated with anesthesia induced in rainbow trout by MS-222 (Kestin et al. 2002). 

Reflex impairment related to pollutant concentrations

Reflex impairment can be used to test for the behavioral and fitness outcomes of exposure to possible pollutants.  A recent study tested effects of triclosan on reflex impairment in an estuarine fish, Atlantic croaker:

"The effects of triclosan on reflex responses and anti-predator behavior in an estuarine fish

Tiffany L. Hedrick-Hopper and Sandra L. Diamond, Department of Biological Sciences, Texas Tech University, Lubbock, TX Background/Question/Methods Triclosan is a common antibacterial compound found in an increasing number of personal care products including toothpastes, deodorants, and soaps. Despite partial removal by wastewater treatment plants, an increasing amount of triclosan is entering watersheds where it can have significant effects on aquatic organisms. Even at low levels, triclosan negatively impacts thyroid homeostasis in anurans and fish, and it can decrease startle responses and activity levels in anurans. The purpose of this research was to investigate the effects of triclosan on reflex responses and anti-predator behavior in juvenile Atlantic croaker (Micropogonias undulatus), an estuarine fish. Sixty Atlantic croaker were held in individual tanks and randomly assigned to be fed a diet of either normal food pellets or pellets impregnated with 50 ppm triclosan for 14 days. Both prior to and immediately following the 14 day exposure, fish were tested for a suite of reflex action mortality predictors (RAMP) and were subjected to a video-recorded 30 second simulated predator attack. Videos were then analyzed for the specific strategies (run, hide, cut across tank, turn gambit) employed by the fish before and after exposure. Results/Conclusions We found that fish exposed to triclosan were significantly more likely than control fish to exhibit reflex impairment. Specifically fish lost the dorsal spine erection response, meaning that they did not raise their dorsal fin when the fin was flattened. Reflex impairment is correlated with increases in overall fish stress and mortality outcomes. Treated fish also experienced significant shifts in their anti-predator strategies. Triclosan-exposed fish spent significantly more time in their post-exposure test hiding from the simulated predator than fish in the control group. In some cases, fish continued to stay stationary even as the simulated predator touched them. The results of this study indicate that triclosan does impair fish reflexes and creates shifts in the strategies used by croaker to escape their predators. Since these schooling fish have been shown to exhibit dominance hierarchies, triclosan may affect social patterning in Atlantic croaker. These behavioral effects may have important implications not only for croaker and similar fish species but also for croaker predators such as bottlenose dolphins as contaminated fish may be easier prey, leading to increased predator body burdens." This study used free-swimming fish and video analysis of reflex responses.  The RAMP approach can easily be adapted to pollutant research and aquaculture settings for efficient, real-time monitoring of supply waters and sediments, as well as health conditions for animal rearing.  In the basic research context, neurobiological studies of zebrafish have made use of reflex testing in pharmacology and toxicology laboratory settings.

Uses for RAMP

RAMP measures reflex impairment and predicts delayed mortality in animals. The method is a real time, cheap, effective, and easy way to monitor animal vitality, stress, disease, morbidity, and delayed mortality.  How is this useful?

In fisheries, non-target species of fish, invertebrates, birds, amphibians, reptiles, and mammals (bycatch) are captured and released for a variety of reasons, usually related to management requirements or economic factors.  The release of these animals can result in large, unquantified amounts of fishing mortality that causes uncertainty in fisheries management and wastes valuable lives and resources.  RAMP can give immediate real time data on the vitality and potential delayed mortality of captured animals.  This data can be used in real time to evaluate and adjust fishing practices to improve bycatch survival and to quantify bycatch fishing mortality.

In live fisheries, animals are captured and transported to net pens or land markets for food consumption or aquarium trade.  Sorting of animals for maximum survival can be quickly accomplished using RAMP.  This sorting saves valuable holding space, decreases shipping costs, and increases the value of catch by including only top quality animals.  RAMP also aids in perfecting capture practices that maximize animal vitality and survival.

In aquaculture, RAMP is useful for real time monitoring of animal vitality, stress, disease, and potential for morbidity and mortality.  Aquaculture depends on maintaining the highest health conditions for animals, as disease can be a major impediment to aquaculture ethics and economics.  RAMP monitoring can be automated in aquaculture settings by using computer aided reflex testing.  Stimuli such as flashing bright light, sound, and food scent can be administered randomly into holding areas and the reflex responses of free-swimming animals can be observed and analyzed by video systems.  Detection of impaired reflex responses can trigger alarms systems and bring personnel to adjust conditions for improved animal health.

In pollution research and monitoring, RAMP is a sensitive measure of animal vitality, stress, sublethal, and lethal effects of pollutants. A large body of research exists documenting effects of pollutants on volitional feeding, social behavior, and predator prey interactions. RAMP can be an even more powerful measure of pollution effects than volitional behavior, since reflex impairment is directly related to animal vitality, without the modifying effects of motivation, size, and sex.