Cover letter to all parties interested in Mexican wolf reintroduction

 

We regret that our proposal is coming so late in the Mexican wolf reintroduction effort.  We believe that we may be able to substantially reduce conflicts between Mexican wolves and domestic livestock using procedure known as conditioned taste aversion (CTA) as described in the following report.

 

We understand that the readers of this report are very diverse in background and interest.  We have prepared it so that anyone reading only from the main text of the report can understand the essence of what has happened and what we are recommending.  For all wishing to examine our claims more carefully, we offer a series of appendices that explain things more closely.  We also provide a list of scientific publications that substantiate our claims, conclusions and recommendations.

 

We hope that all of you will take a little time to consider this possible solution to this problem and support an effort to test it.  With the minimum required support, we are willing to do what is necessary to see to it that a good first effort gets under way, to be sure that the effects of this effort are openly and objectively measured and that personnel are adequately trained in this new procedure.

 

We do not consider ourselves the final source of wisdom on this subject and so please send us your suggestions and comments.   We will respond quickly and fully if you just say who you are.   We will not respond to anonymous persons.  It may well be possible that someone might suggest something that is missing from the report or that ought to otherwise be changed and so its final form and what is ultimately done might be changed somewhat from what we are presently submitting.

 

Thanks,

 

Lowell Nicolaus, Contact: wolfishnicolaus@earthlink.net

Dan Moriarty, Contact: moriarty@sandiogo.edu

 

FINAL REPORT: A CONDITIONED AVERSION PROTOCOL FOR MITIGATING CONFLICTS BETWEEN LIVESTOCK AND WOLVES DUE TO BE RELEASED INTO LIVESTOCK HABIAT

 

Lowell K. Nicolaus, Ph.D.

Emeritus, Northern Illinois University

And

Daniel D. Moriarty, Ph.D.

University of San Diego and the California Wolf Center
 

Introduction

Mexican wolves were reintroduced into the Blue Range Wolf Reintroduction Area of Arizona and New Mexico in 1998.  Then, wolves began to prey on livestock in the region, averaging 20 depredations of cattle per 100 wolves per year.  Of course this led to conflicts between those advocating reintroduction of the wolves and cattlemen with their livelihoods at risk.  Livestock depredation has led to control action including lethal removal of wolves.  The Interagency Field Team has also employed some non-lethal methods to discourage wolf depredations upon domestic livestock including hazing of wolves and fladry barriers but the depredations continue. 

Here we report and discuss the results of some recent research using a process known as conditioned taste aversion (CTA) that suggests a powerful alternative method for reducing conflicts between Mexican wolves and domestic livestock.

It has long been known that when humans and animals experience a severe gastrointestinal illness after consuming food, the taste and scent of this particular food become disgusting even when the food was earlier very preferred and familiar.  In humans, this has been shown, for one example, among cancer patients who consume food just before the onset of nausea induced by chemotherapy or radiotherapy.  Long after recovering from the nausea caused by the cancer therapy, patients respond to the smell and taste of this food with pronounced disgust, often retching and vomiting at the mere scent of the now offending food.  These aversions result after just a single illness and can be extremely persistent.   So, before this process began to be prevented by physicians with understanding of what was happening, patients that underwent repeated chemotherapy treatments often formed aversions to each of their favorite and most nutritious foods in sequence until loss of body weight became a critical factor in their cancer treatment.  Paradoxically, these people can intelligently discuss the fact that the chemotherapy is what made them ill and not the food, but they still gag and wretch rather than to swallow the food and often vomit at a distance based only on the food smell (Appendix 1).

Conditioned taste aversion has been tested experimentally upon bird and mammal predators both in captivity and in the field (Appendix 1).  The procedure usually requires that predators consume a meat bait with the same taste and scent as of live target prey, but laced with a hidden dose of some substance that induces a severe but temporary illness after it is consumed.  If the illness-inducing substance is successfully hidden in the bait, then the predator forms an aversion to the taste and scent of the target prey rather than to the taste and scent of the aversion agent.  When predators consume poorly constructed baits that taste and smell different from live prey, they may begin to avoid the baits but nevertheless will continue to attack and consume live prey (Appendix 2).

Paradoxically even though a well constructed meat bait that induced the illness was inanimate, predators nevertheless tend to be dissuaded from live prey with the same taste and smell as the bait because, like their human counterparts, that taste and smell has become disgusting to them.

We have recently demonstrated the potential usefulness of Thiabendazole (TBZ) as an aversion agent among captive wolves.  The focus of this work was to develop a clinical protocol that safely and effectively produces aversions to the taste and scent of target domestic livestock among captive predators before they are released into livestock habitat.  But with modification, this same procedure could also be used to produce aversion in other applications.  These include producing aversions among recently caught predators while they remain in captivity and could readily be extended to the treatment of free-ranging predators. It could also be used to reduce conflict by inducing aversions to the taste and scent of livestock carcasses.  In all cases, the intent is to produce a disgust for the scent and taste of target prey so that rather than being attracted at a distance to these prey, predators are dissuaded from approaching and conflicting with livestock at a distance.  That is, we expect the critical events affecting depredation upon target livestock to take place at night several kilometers down wind from occupied pastures without any human witnesses when predators choose not to move in the direction of the cattle because for them the scent has become disgusting rather than attractive.  We expect that since the taste and scent of cattle will have become disgusting to treated wolves, wolf and calf will seldom come nose to nose. 

All recently released Mexican wolves can be tracked with radio collars while others already in the field are monitored and wolf depredations upon cattle are also recorded.  As long as these efforts continue in this situation and those with successful experience in the use of CTA see to it that properly constructed baits are distributed and monitored as needed, it ought to become clear whether or not our attempts to produce aversions among wolves reduce livestock conflicts.  The process of producing aversions is relatively inexpensive, can do no harm and at this time there is no better option.

Thiabendazole as a Suitable Aversion Agent
 

Baits designed to produce conditioned aversion among wolves must contain an appropriate dose of some substance that safely but effectively induces a temporary illness after predators consume the bait.  Many substances are known to induce illness but few have the particular combination of features required for an effective aversion agent.  To function as we wish it to, an aversion agent must fulfill all of the following requirements:

·        It must induce an illness severe enough to produce a strong aversion to the taste and scent of the target food. 

·        This illness must commence some time after the bait is fully eaten so that the full, intended dose of the substance is consumed before illness onset terminates the meal.

·        The substance must be safe enough so that when the intended dose is consumed the predator fully recovers after a relatively short time.  And, if there is a possibility that any one predator might consume a greater amount of the aversion agent, the safety margin between an illness dose and a lethal dose must be broad.

·        The substance must have the physical capacity to be adequately mixed in the bait material and remain stable in the bait long enough to remain effective by the time it is consumed. 

·        Importantly, the taste and scent of the substance must not alter that of the bait in any detectable way.  Poorly constructed baits that induce illness but that do not taste or smell like live prey will themselves be avoided by predators that acquire an aversion to them, but predators will still attack and consume live prey that do not taste and smell like the baits they ate.
 

Thiabendazole (TBZ) appears to meet all of these requirements. 

··  Although it might not be the most effective aversion agent in terms of the nature and severity of illness it induces, it has produced clear and lasting food aversions among Black bears, wolves, dingoes, foxes, hunting dogs and other predators (Appendix 3). 

·  The physiological effects of TBZ appear to begin some time within about an hour after a suitable dose is consumed and so this delay should provide adequate time for a full meal to be consumed before illness onset reduces willingness to eat (Appendix 4). 

· TBZ has a very long history as both a human and a veterinary pharmaceutical and as a fungicide applied broadly upon human foods.  In these roles, its toxicity and safety are documented by a very extensive literature demonstrating its safety (Appendix 5). 

·   As an organic compound, it mixes easily in meat baits containing animal fat and appears to remain stable in meat baits for a sufficient time to remain effective under field conditions.  In fact, the baits used in the current study were manufactured in the state of Oregon and frozen for several months before they were used at the California Wolf Center.

·  Most important, although it is probably not fool-proof and so should be mixed very carefully in meat baits, TBZ does not appear to have a strong scent or taste of its own and so it can be added to food in recommended amounts without enabling predators to discriminate between the all-important taste and scent of bait and that of live prey (Appendix 2).

A DEMONSTRATION OF THE CONDITIONED TASTE AVERSION PROTOCOL

 
The Wolves and the Setting

The procedure reported here was demonstrated at the California Wolf Center (CWC) near Julian, California over the month of September, 2008.  A pack consisting of 9 Alaskan gray wolves (Canis lupus), including 6 males and 3 females, was maintained in a common outdoor enclosure approximately 30 meters (90 feet) on each side.  All of the pack members had been raised in captivity and until shortly before being moved into this relatively restricted space to facilitate observation, had been maintained together long term in a much larger portion of the facilities at CWC in addition to the enclosure within which they were recently limited.  Three of the wolves were old and to some extent crippled by injuries and age.  The remaining pack members ranged from two years of age to 8 years of age.

The enclosure had clean water available at all times and shade trees were dispersed largely on the sloping upper half of the space.  With the exception of one low area surrounded by logs, nearly all of the interior of the enclosure was visible from the outside.    No objects or devices were provided in the enclosure for the purpose of enriching the environment for the wolves. 

Prior to the start of the demonstration, in order to minimize strife and stress, the wolves were fed a ration of meat and/or fish at a rate of almost 8 Kg (20 pounds) per wolf in each of two weekly feedings.  Each of the weekly totals of 16 Kg was more than the wolves ordinarily consumed immediately and so there was usually food left from earlier feeding at all times and so the feeding regimen was essentially ad lib. 

The wolves were largely free of immediate human contact other than the brief occasions when volunteers and staff entered the enclosure to feed them and to remove obvious articles of offal.  Visitors to the CWC were required to remain in an observation area approximately 30 m from their enclosure and warned not to make noise or otherwise threaten the wolves.

The Sequence
 

The essential logic of this protocol is that wolves must voluntarily consume illness-inducing baits whose exterior and interior maintain the unique taste and scent of live target domestic livestock prey.  After fully recovering from the effects of the aversion agent, they must then demonstrate that they have acquired an aversion to the taste and scent of target prey before being released into habitats where they may conflict with livestock. 

 

Food Deprivation.  The protocol requires that wolves be subjected to a feeding regimen including sufficient food deprivation to induce them to voluntarily participate in the protocol.  This is because they must voluntarily consume baits that, because they may initially be unfamiliar to them, might not be readily eaten and includes times when they are offered treated baits that they must fully consume in order to acquire an aversion.  Food deprivation should induce predators to consume their maintenance ration when offered to show that they are fully recovered from the effects of the aversion agent.  Finally, deprivation provides assurance that wolves that refuse baits during the post-test after acquiring an aversion are doing so because of the aversion rather than because of satiety.

 

In order to achieve the food deprivation we needed to apply the protocol, we fed the wolves every other day for 4 days per week and completely withheld food on the intervening 3 days per week.  When fed, each received only 3 Kg (about 6.5 pounds) in a single meal lasting several hours. These feedings took place several hours after our morning trials when we introduced baits designed to induce aversions.  The greatest difference between the old and this new feeding regimen was that few hours after each feeding, we removed all of the uneaten food and so we completely food deprived the wolves between feedings.  In this way, 4 days per week before each morning trial we had completely food deprived the wolves for approximately 20 hours and on three intervening days per week we deprived them for over 40 hours before the morning trials.  This was the first time that the wolves had been subjected to a feeding regimen that included any food deprivation but they withstood it well without evidence of weight loss or of increased strife.

 

The protocol should include an initial pre-test; a period of time when predators consume meat baits with the taste and smell of target livestock but without the aversion agent. This achieves several things.  Predators allowed at first to consume baits without the aversion agent become familiar with the taste and scent of this food and so are more willing to fully and quickly consume treated baits that deliver the intended dose of TBZ when offered a little later.  The pre-test also demonstrates whether wolves are normal candidates to participate in the aversion protocol.  Obviously, wolves unable or unwilling to consume target prey in the normal manner during the pre-test are probably poor candidates.  The pre-test should provide opportunity to adjust procedural details so that wolves fully consume uncontaminated baits during treatment and can be fairly tested during the post-test.  Since under captive conditions, wolves often display idiosyncratic behavior, the pre-test provides an individual history of the behavior of each wolf that can be used to compare with post-test behavior in determining the extent to which each wolf has acquired an aversion during its treatment period. 

 

The protocol must include a period of time during which baits with the same external and internal taste and scent cues as those consumed during the pre-test deliver a dose of hidden aversion agent sufficient to induce the requisite physiological effects that produce the aversion.  The aversion agent should be mixed within the inner meat of the bait such that its taste and scent are not detected by the consumer.   

 

The protocol requires that treated wolves demonstrate that they are fully recovered from the effects of the aversion agent before being tested to establish whether or not they have acquired an aversion.  This can be accomplished only by offering them their ordinary maintenance ration 6-8 hours after they are treated with TBZ.    After wolves have fully recovered, it requires that untreated baits again be offered  in order to document that predators now avoid the outer surfaces of baits on the basis of taste and scent before each wolf is released.

Our demonstration of this protocol was therefore divided into three phases: a 4-day pretest, a 10-day treatment period and a 4-day posttest.   During the pretest, for 4 consecutive mornings beginning at first morning light we threw two untreated sheep meat baits over the enclosure wire, one at a time (Appendix 6).  The wolves were allowed to interact with the first bait for 60 minutes before the second bait was provided for a second 1-hour period.  Each of the baits consisted of 100 g of plain ground mutton without TBZ wrapped with fresh, clean sheep hide.  The hide was securely sutured with double square knots with double cotton thread at intervals of 1 cm (0.5 inches) along each edge of the bait package.  In this way, we intended that wolves be exposed to the unique exterior taste and scent of sheep by being forced to spend substantial time and effort chewing their way into the bait to get to the interior meat.  During each of these hour-long episodes, we remained outside of the enclosure and as unobtrusively as possible video taped all of the interactions during each period.  At the end of this time, we entered the enclosure briefly to remove any unconsumed portions of the baits to be examined and weighed.

This was immediately followed by a treatment period of 10 consecutive mornings during which wolves were offered one TBZ treated bait for 60 minutes and we recorded all events as before.  The TBZ treated baits consisted of 250 g of ground mutton with 5.4 g of TBZ well mixed with the meat.  This TBZ treated meat was then securely wrapped with fresh, clean sheep hide and sutured closed as before.   At the end of each of these episodes we entered the enclosure to recover all remains of the baits. 

Since the average body weight of the wolves was estimated to be 38.5 Kg (98 pounds), an absolute amount of 5.4 g of TBZ would deliver an average dose of 140 mg TBZ/Kg wolf body weight.  But some of the largest males weighing about 50 Kg (125 pounds) would receive a dose of TBZ as little as 113 mg/Kg and some of the smallest females that weighed only 27 Kg (69 pounds) would receive 200 mg TBZ/Kg their body weight.

Finally, there was a posttest composed of 4 consecutive days during which just the same procedure including untreated meat baits as used for the pre-test was repeated. 

Analysis

We scored the video record of wolf interactions with the baits to characterize any changes in the frequencies of common predatory/feeding behaviors that have been used in earlier studies of animals with food aversions and that might be of use in a practical effort to evaluate whether or not captive predators had actually acquired an aversion to the taste and scent of baits prior to their being released.  Our statistical analysis of the database was the simplest, most intuitive, most conservative, and least dependent upon statistical assumptions that we could logically use.

 

RESULTS
 

Safety of Thiabendazole

Of the four wolves in the pack that fully consumed a TBZ-treated bait, two were small females that each received a dose of approximately 200 mg TBZ/Kg their body weight.  The other two were larger males receiving relatively less of a dose.  Four other wolves that did not consume a full bait received less TBZ still.  Since only one bait was offered each day during the treatment period,  each of the TBZ treated wolves received treatment on separate days during which we could monitor their recovery.  Since the symptoms of TBZ treatment are mainly just lethargy, after consuming the bait each of the treated wolves found a place in shade to lie down and sleep.  At about this time of the day with rising temperatures the other wolves of the pack found shade for sleep also.  As a result, no wolf that was treated was molested by any of the other wolves while potentially disabled under the influence of the TBZ.  All fully treated wolves consumed their maintenance ration in the normal fashion when it was offered approximately 6 hours after they consumed TBZ.  Consequently, we concluded that the physiological effects of this dose of TBZ persisted less than 6 hours, but cannot say precisely when recovery occurred before the 6 hours when they were fed.  Aside from changes in willingness to approach and consume baits subsequent to this, there were no changes in the social rank or physical capacity of any of the fully treated wolves of the pack.

Effectiveness of This Demonstration (Problems)

Because we were obliged to conduct this demonstration under the prevailing conditions at the CWC where wolves had to be maintained all together in a common enclosure, we identified three problems that must be avoided in the future.  These were:

·  Bias.  Our presence near the enclosure and our having to throw the baits over the enclosure wire introduced a bias strongly favoring the least shy members of the pack.  Of the nine wolves in the enclosure one, Kuma, was a large male so shy of humans that none of the volunteers or staff at the CWC had ever seen him consume food in their presence.  Though he paced broadly and actively, he never interacted with baits throughout the demonstration and so will not be mentioned further.  Naomi was a shy low-ranking female who eventually consumed treated bait but did so under such cover that we could not say precisely how much she consumed.   On the opposite spectrum, Taku, a young low ranking male, and Minka, a potential alpha female, were the least shy and tended to dominate the baits.  Typically, Taku first seized a bait, dragged it around and played with it before opening it, often spreading the interior meat over the ground and urinating on it before higher ranking individuals took the bait. Taku never consumed a full treated bait and so although he stopped consuming any of the bait material, he continued to play with the baits, contaminate the outer surfaces of baits and spread the meat onto the ground. Until she finally consumed a full treated bait, Minka also seized baits but this ceased after her treatment.  Because of these interactions, many wolves that might have fully consumed treated baits without contamination of the taste and scent of the exterior merely scavenged portions of meat from contaminated baits.

·  Contamination.  Our intention was for wolves to consume baits whose outer surfaces retained the unique taste and scent of target livestock.  Requiring wolves to chew their way through the hide was to force them to be exposed to these taste and scent cues before illness that followed eating treated interior meat.  But, the exterior taste and scent of all of the baits were both substantially altered by at least three different sources of contamination.  No bait escaped being urinated upon at least once and sometimes several times and by up to three or four different wolves.  This occurred before, during, and after baits were opened.  Wolves also often rolled over and rubbed their faces onto the surfaces of baits and in so doing, probably imparted their particular scent gland secretions.  Finally, the ground of the enclosure was a very fine, almost powdery dry material containing the urine, dried fecal material and decomposed offal that had accumulated over many years.  All food, including all of the baits, became thoroughly coated with this highly familiar and highly scented residue and so the exterior of all of the baits became much less distinguishable from everything else that the wolves consumed as food.  The result of all of this was that no wolf had a single, clear exterior taste and scent cue before becoming ill from eating treated meat and so the exterior of baits did not become a barrier to some of the wolves.

·  Under Dosing.  Several wolves competing for baits resulted in treated meat being broadly spread and so scavenging resulted in 4 of the wolves receiving only partial treatment with TBZ.

Because of these difficulties, none of the baits remained unopened by wolves throughout the entire protocol.  Instead, while fully treated wolves avoided baits largely at a distance, partially treated wolves opened bait exterior with the altered taste and scent but they rejected the interior meat, ending up spreading it all over the enclosure.  On two occasions during the pre-test, empty hides composed of belly hide were thin enough to enable wolves to chew them enough to be swallowed.  All other hides were left for us to recover after tests.

Though problematic in this particular demonstration, it is well that these difficulties alerted us to what might take place under similar circumstances and this greatly contributed to our certainty concerning what to recommend for future efforts. 

Effectiveness of Thiabendazole (Measures of Effect)

During the pre-test, all but one of the wolves that ultimately participated in the protocol partially or wholly consumed at least one untreated bait. We observed no hesitation on the part of any of the wolves to approach, open and consume uncontested baits.  The absence of any evidence of neophobic behavior (the tendency to treat new foods with suspicion) toward this initially unfamiliar sheep meat may have been partially due to the fact that the wolves at CWC were fed an extremely varied maintenance diet and so may have become accustomed to novel foods (Appendix 6).  On no occasion at this time was anything left other than the hide after the first 30 minutes.  The usual pattern was for a bait to be seized, opened with the use of all of the complement of teeth and the interior meat quickly consumed either all at once or scavenged when it was spread around.  Other than the fact that Kiana and Taku tended to be the first to seize the baits during the pre-test, there was no other obvious pattern indicating that the first bait in any way predisposed which wolf might open and consume the second bait. 

 

Even under the unfavorable conditions described above, treated wolves demonstrated the most common indications of having acquired an aversion to the baits (Appendix 7).  One of the most obvious changes in behavior attributable to treatment with TBZ was an absolute suppression of the willingness of these predators to consume the baits once each of them had been treated (See Table 1 below).  Four of the wolves fully consumed a bait and then abruptly stopped consuming bait.  So, in their case the amount consumed and TBZ dose received was certain.  The other four wolves scavenged broadly distributed meat.  At these times our estimates of amounts consumed by each wolf were based upon careful analysis of the video record and the recorded weights of bait material recovered from the enclosure after each trial. 

 

We reentered the enclosure to collect all of the remains of baits that we could find, but it is certain that we did not recover all unconsumed bait.  Some of the meat, for example, was mixed into the enclosure soil and/or divided into extremely tiny pieces.  In addition, the meat attracted swarms of carnivorous wasps, each of which removed meat so that in the aggregate we might have lost appreciable amounts within the hour before be recovered the meat.  The amount of recovered inner meat indicated below was the aggregate per cent of all that was offered for that day (100g offered twice during each of the pre-test and post-test days, 250g during each of the treatment days).  Hides varied according to thickness and amount of material they accumulated in the enclosure and so were not weighed.

  

 

        Table 1. Consumption record of wolves participating in the 

                      Thiabendazole aversion protocol at CWC.

 

Wolf                             PRE-TEST                               TREATMENT                                    POST-TEST

                               1       2      3       4          1     2     3     4     5     6     7     8     9     10          1     2     3     4                      

FULLY-TREATED
WOLVES

Kiana                   YES   YES  YES   YES        100%  [NO FURTHER CONSUMPTION…………………………………..…...]

Minka                       YES                      100% [NO FURTHER CONSUMPTION…….…………..……...…]
 

Ketchikan                       YES  YES                 100% [NO FURTHER CONSUMPTION………….…..…]

 

Barrow                               YES                        97% [NO FURTHER CONSUMPTION……….….]

     

PARTLY-TREATED

WOLVES 

Taku                             YES  YES  YES              20%          30% [NO FURTHER CONSUMPTION……….…...…….…...]

 

Inuk                              YES  YES                       15%          30% [NO FURTHER CONSUMPTION…….….……….……..]

Tundra                                 YES                                           30% [NO FURTHER CONSUMPTION………….….….……..]

Naomi                                                                                                          20%

RECOVERED
MEAT (%) 
      0         0        0        0           0      60     0     10      0       3      90      20     60     80            75      99     90     75

 
Inspection of the above data reveals several important features of this process:

· By mid-way through the treatment phase of the protocol, all of the wolves destined to be treated other than Naomi had been treated and ceased consumption of baits.  As expected for a successful aversion, the most active predators tended to be the first treated and these wolves simply left TBZ meat to be consumed by others in their own turn.

· Those that had become most familiar with the baits during the pre-test were as likely to cease consumption of baits after a single full treatment or partial treatments as those least familiarized with baits during the pretest.  Therefore, familiarity with prey did not seem to impair the acquisition of an aversion to it.

· Wolves that were treated early underwent up to 17 subsequent post-tests (including 2-1 hour trials during each of the 4 post-test days) during which no food other than the bait was available and on almost half these occasions after having been food deprived for over 40 hours.

· Of the wolves that received only partial treatment with TBZ, two consumed some of the bait meat twice and then ceased eating baits, one ceased consuming bait meat after only a single incomplete dose.  This indicates that even partial treatment can have some effect upon food preference though partial treatment might not produce lasting and robust effects for wolves destined to be released.

· Once treated with TBZ, wolves continued to refuse baits in spite of the near and continuous presence of other untreated wolves actively consuming baits. 

· Since all of the wolves continued to reject the interior meat of baits during the post-test when no TBZ was present in the bait, it appears that the wolves failed to detect the TBZ in the bait during the treatment period.  That is, instead of forming an aversion to the TBZ, the illness induced by TBZ was associated with the taste and scent of the meat itself.

In addition to complete suppression of actual consumption of bait contents, wolves demonstrated a change in willingness to approach the baits from a distance.  Two obvious measures of this tendency were identified on the video record and have been used before as measures of aversion (Appendix 7).   We assumed that it was most unlikely that any wolf that moved within 1 meter (three feet) of any available bait but then continued past the bait without any observable hesitation had done so because it did not know the bait was present.  Wolves that engaged in this behavior were therefore scored as having “ignored” the bait.  Another behavior was the tendency to reject the bait on the basis of its scent.  In this case, wolves were scored as having “Smell-Rejected” a bait if they came head-on toward a bait within 1 meter, hesitated with the nose near the bait, and then chose to move on without making physical contact with the bait.  The total frequency of these two behaviors is reported on Table 2.

    

 

 Table 2. Frequency of avoidance of baits at a distance (Ignoring baits and smell-rejecting baits).

   

          Wolf                           PRETEST                   POST-TEST

FULLY-TREATED
WOLVES

Kiana                                          0                            46

-------------------------------------------------------------------------------------

Minka                                                  0                                     4                            

-------------------------------------------------------------------------------------

Ketchikan                                          0                                    35

-------------------------------------------------------------------------------------

Barrow                                                0                                    21

-------------------------------------------------------------------------------------

TOTAL                             0                            106

------------------------------------------------------------------------------------------------------

PARTLY-TREATED
WOLVES

Taku                                            0                           42

-------------------------------------------------------------------------------------

Inuk                                                     8                                    34

-------------------------------------------------------------------------------------

Tundra                                         0                             21

-------------------------------------------------------------------------------------

Naomi                                                  3                                     2

-------------------------------------------------------------------------------------

TOTAL                              11                          99

  

The number of wolves that increased avoidance of baits at a distance after treatment compared with before treatment was seven out of eight.  Although it is very unlikely that very hungry wolves without a food aversion would otherwise behave in this way, we arbitrarily assign the likelihood of hungry wolves ignoring potential food as 50% (1 time out of two) for each wolf. The computed likelihood of getting 7 out of 8 is actually less than one chance in 100 trials (Sign Test, P=0.50, Alpha<0.01).  This is exactly the likelihood of getting 7 heads out of 8 tosses of an honest coin.  Therefore, avoidance of these baits without aversion is very unlikely and so the behaviors above are reliable and practical measures of whether or not predators have acquired an aversion before being released from captivity.

This is not to say that there were no occasions during the post-test when treated wolves entirely avoided baits at a distance.  In fact, wolves did occasionally proceed to taste baits since contamination of the outer surface prevented it from becoming the unambiguous barrier that we intended.  By far, the most frequent rejection of baits on the basis of taste took place with the two young wolves Kiana and Taku who continued to seize baits when they were offered. Nothing in the enclosures enriched the wolf environment and so the wolves were undistracted except upon occasions when wind blew objects into the enclosure or a hapless Raven was caught by a wolf that was then pursued by others before the Raven was finally dismembered and left uneaten. CWC staff remarked that the behavior of the two youngest wolves, Taku and Kiana, appeared more to be curiosity and playfulness than feeding behavior since they tossed baits into the air and rolled playfully over them before attempting to open them.  They spent much more time to open the baits than they did before treatment since they now used only their incisors and took time with exaggerated mouth and tongue movements in attempts to remove individual pieces of hide from their mouths.  This accounts for the high frequency of recorded “Taste-Reject” behavior.  On every occasion for every wolf the taste of the hide and that of the interior meat was rejected instead of being followed with normal consumption (See Table 1).  When the wolves encountered the taste of interior meat, they shook their head, gagged, or stopped contact and simply left the bait.  Occasions when wolves made mouth contact with hide or meat without going on to consume the bait were scored on the video record as “Taste-Reject” behavior as reported in Table 3.

Table 3. Frequency of rejection of baits based upon taste.

Wolf                                        PRE-TEST                          POST-TEST

FULLY-TREATED
WOLVES

Kiana                                         0                                             23

-------------------------------------------------------------------------------------------------

Minka                                             0                                               1

-------------------------------------------------------------------------------------------------

Ketchikan                                       0                                               1

-------------------------------------------------------------------------------------------------

Barrow                                           0                                               8

-------------------------------------------------------------------------------------------------

Total                                               0                                              33

    

PARTIALLY-TREATED
WOLVES

Taku                             0                                          31

--------------------------------------------------------------------------------------------------

Inuk                                                0                                               5

--------------------------------------------------------------------------------------------------

Tundra                                           0                                                8      

--------------------------------------------------------------------------------------------------

Naomi                                            0                                                0

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Total                                              0                                               44

Again, it is extremely unlikely for any hungry wolf to reject the taste of potential food unless they have acquired some kind of an aversion to the food or the food is laced with some kind of noxious substance.  Nevertheless, if we set the likelihood of this kind of behavior being observed by chance as 0.50 (50% or 1 out of every 2 trials) as before, the chances of 7 out of 8 individuals engaging in this behavior during the post-test is very unlikely indeed.  The probability of this occurring is much less than 1 occasion in 100 trials (Sign Test, P=0.50, Alpha <0.01).

 

Please note, however, that although “taste-rejection” behavior is very dramatic when it occurs, it only happens when aversion on the basis of the external scent of prey is insufficient to produce avoidance at a distance.  This could occur when wolves receive an insufficient dose of aversion agent, detect the aversion agent in the bait or when the external taste and scent cues had become confused or obliterated. 

 

Conclusions

 

Our demonstration of this protocol assures that wolves can be induced to voluntarily participate in the full protocol from pre-testing through post-testing.  In this protocol, familiarity with target prey during the 4 days of the pre-test did not prevent a single average dose of Thiabendazole of about 140 mg TBZ/Kg wolf body weight from producing clear aversion to the taste and scent of the inner meat. This was confirmed through practical measures of behavior changes that have been used to measure aversions before.  It seems clear that wolves failed to detect TBZ when well mixed in inner ground meat at a rate no greater than 2-3% of the weight of the meat.  It is equally clear that efforts must be made to eliminate the influences of bias, contamination and under-dosing that affected the results of this demonstration.  If wolves do not consume a full dose of TBZ or if the outer surfaces of baits are contaminated so that the unique taste and scent of target livestock prey is masked, reduced,or confused then there is little hope that an aversion acquired under captive conditions will reduce predation on livestock.  We offer some recommendations for future use of this protocol (Appendix 8).   We hope that the history of substituting poorly conceived procedures that produce failures rather than simply following procedures known to produce desired outcomes will not be repeated (Appendix 2).

 
APPENDICES
 

Appendix 1.  On their web site, Riley and Freeman (see literature cited section below) list over 3000 research articles describing CTA.  Many of these are clinical studies dealing with the treatment of alcoholism, food preferences of humans and animals, treatments for weight loss or gain and many other subjects in addition to wildlife management.  The first studies reporting human cancer victims forming dramatic and life-threatening food aversions resulting from chemotherapy were reported by Bernstein, 1978; Bernstein and Webster, 1980; and Bernstein, 1985.

 

Successful research on CTA in which predators consumed carefully constructed baits designed to produce illness began with Gustavson who first reported that CTA could produce avoidance of live prey among captive wolves and coyotes (Gustavson, et al. 1974; Gustavson, et al. 1976).  He later reported that meat baits placed into the field reduced predation by coyotes upon sheep (Gustavson 1976).  These results were later confirmed independently by Ellins, et al. 1977 and by Stream 1976.  Gustavson again reported comparable results in Canada (Gustavson, et al. 1982).  Nicolaus, et al. 1979 and Nicolaus, et al 1982 confirmed that aversions induced by inanimate baits could inhibit predatory attack upon live prey.  But this was accomplished in the field rather than in captivity.  Individually identified free-ranging raccoons that had consumed chicken carcasses carefully laced with an aversion agent later avoided live chickens as well as the carcasses.  Later, Gustavson and Nicolaus, 1987 summarized all of the research on this issue up to that date.  Over time, many studies both in captivity and in the field have confirmed that predators quickly and dramatically alter predatory behavior when affected by CTA.  In addition to research mentioned above, CTA has been shown to be effective when baits have been distributed in the field to treat free-ranging predators such as raccoons (Semel and Nicolaus, 1992), black bears (Polson, 1983; Ternent and Garshelis, 1999), mongooses (Nicolaus and Nellis, 1987), whole assemblages of predators converging to prey on the same prey species (Nicolaus, 1987; Nicolaus, et al., 1989), Common Ravens (Nicolaus, 1987), American Crows (Nicolaus, et al. 1983; Nicolaus, et al. 1989; Dimmick and Nicolaus, 1990) and Redwing Blackbirds (Nicolaus and Lee, 1999).  In addition to this, a number of other researchers have reported successful establishment of CTA among captive predators.  These include Gill, et al 1999; Massei and Cowan, 2002; Massei, et al., 2003; Massei, et al., 2003 and a good summary of research in field and captivity was produced in 2000 by Cowan, et al. 

Appendix 2. There is very good reason to believe that when applied properly in meat baits, predators do not detect the taste or scent of the TBZ in the bait.  In the successful studies cited in Appendix 1 above in which TBZ was used as the aversion agent, TBZ was variously added to such foods as meat baits, bee’s wax and honey, and in military rations.  Since care was exercised to use carefully measured amounts well mixed with a large enough amount of the food to mask its odor and taste, TBZ induced aversion to the taste and scent of the food such that during post testing, predators avoided the foods without the TBZ being present.  An extensive literature reveals what happens, however, when baits are constructed so that the aversion agent systematically alters the taste and scent of the bait.  Naďve and hungry predators often willingly consume the baits but since the taste and scent of the bait do not match that of live prey, they readily discriminate between baits which they are no longer willing to consume and live prey that do not taste or smell like baits that induced the illness (see: reviews of this literature in Gustavson and Nicolaus, 1987; Bekoff, et al 1975; Ellins, et al 1978: Quick, et al 1985; Nicolaus at www.conditionedtasteaversion.net)

Appendix 3. TBZ produced clear and lasting food aversions among free-ranging black bears when applied to the problems of reducing depredations upon bee hives in Canada (see: Polson, 1983) and in bear-human conflicts in a military reservation in Minnesota (see: Ternent and Garshelis, 1999).  After a close analog of TBZ, Mabendazole, was seen to induce food aversions among young wolves, TBZ itself was used to produce aversions among Dingoes and Hunting Dogs (see: Gustavson and Nicolaus, 1987).  Later, TBZ was used to produce food aversions among foxes and other predators (see: Masssei and Cowan, 2002; Massei, et al, 2003).

Appendix 4. Some substances like Lithium Chloride produce very obvious illness symptoms including gagging, vocalizations and vomiting and so the time for the onset of illness is usually obvious.  The physiological effects of TBZ, however, are far more subtle and so the onset of its effects is best judged by when predators treated with TBZ begin to refuse food.  That is, animals beginning to become ill cease feeding.  When such a thing has been indicated in the literature, the time appears to be about 60 minutes (see Ternent and Garshelis, 1999; Gustavson and Nicolaus, 1987).  For the same reasons, animals can be judged to have recovered from the effects of TBZ when they first are willing to consume alternative food normally.

Appendix 5. TBZ is 2-(4-Thiozolyl) benzimidazole.  It has been approved for many years as both a human and a veterinary pharmaceutical acting as a broad-spectrum preparation principally for removal of gut worms (see: Merk Index, 1968).  Since the body weight of humans and animals largely determines the effects of toxins, pharmaceuticals and nutrients, the dose of such substances is expressed as so many milligrams (mg: meaning thousandths of a gram) of the substance per Kilogram (Kg: meaning thousand grams) of the human or animal body weight.  The dose of TBZ necessary to kill 50% of test rats is 3100-3600 mg per Kg rat body weight, 3810 mg/Kg in mice and 3850 mg/Kg in rabbits (see: the Veterinary Drug Handbook, 1991).  These doses are over 19 times greater than the 200 mg/Kg maximum dose we propose to use with wolves. Dogs have been force fed doses of 200 mg TBZ/Kg their body weight daily for a full two years without any observable ill effects.  TBZ has been approved and used extensively as a broadly applied fungicide applied to human foods with a very good safety record.  The only environmental hazards so far identified in this agricultural use involve potential toxicity to shellfish and other marine animals when quantities of it wash into the sea.  As long as TBZ remains in baits that are then disposed of in a proper manner, none of it should ever escape into the environment at all.

Appendix 6. We do not recommend that in future baits continue to be introduced to wolves in this manner.  We initially attempted to introduce the baits in a more systematic, perhaps less biased manner but since the wolves were hungry, there was a risk to the CWC staff entering the enclosure to place the baits.  For safety concerns we opted to introduce the baits in this way and admit the potential bias.

The maintenance diet for wolves at CWC was varied, including chicken, fish, road killed deer and other game, horses and mules volunteered by well-wishers.  It was important that we choose a target prey that was unlikely to be donated to the CWC to help feed the wolves.  Since very few sheep were raised in the vicinity, this food had not been fed to the wolves at CWC for many years and so we considered that before we familiarized the wolves with it during the pre-test, sheep was unfamiliar to them and that once established, aversion to sheep would not materially affect their future maintenance.    

Appendix 7. Before acquiring an aversion, hungry predators typically approach, smell, taste, and then fully consume familiar and preferred foods.  The 4-day pretest functioned to demonstrate that the wolves engaged in normal feeding behavior and it also made the baits familiar enough to the wolves so that they might more readily consume treated baits as required.  In nature, consuming nutritious prey provides an internal reward that reinforces external feeding behavior.  That is, successful feeding on a particular food enhances the attractiveness of the taste and scent of the food, directing feeding behavior in a way that increases likelihood of successful feeding.  In nature, when the consequence of consuming prey with chemical defenses is a severe internal illness rather than an internal reward, the taste and scent of the particular food consumed before the illness becomes disgusting.  Rather then to continue to be attracted by this prey at a distance, predators are dissuaded and so seek other, more suitable prey. 

 

Systematic studies of illness-induced change in the attractiveness of the taste and scent of prey have identified several very common and practical measures of this change that should be applied to judging at the end of the proposed protocol whether or not a predator has acquired an aversion before it is released.  These well known behaviors shown by predators with prey aversions include “ignoring” or “smell-rejecting” target prey from a distance and “taste-rejecting” foods that they allow to enter their mouths.  Often “taste-rejection” behavior is complete with head-shaking, gagging and even vomiting at the taste of the now offending food.  We simply used these standard measures of aversion as the criteria for determining whether wolves in this demonstration had acquired an aversion. 

 

Appendix 8. Recommendations 

 

Please note: The Mexican wolf recovery team may be in a position to monitor wolves and their conflicts with domestic livestock.  None, however, have knowledge or practical experience with the successful use of CTA.  Therefore, we consider it absolutely essential to a successful first effort to use CTA to mitigate conflicts between Mexican wolves in each of the applications described below that the work proceed with the guidance of persons with knowledge and experience in the use of CTA.  In each of these three applications, this should include guidance and supervision in the construction of baits, administration of baits whether in captivity or in the field and the monitoring of the fates of baits and behavior of the wolves.  We understand that incidences of conflict between wolves and livestock are documented carefully and in a standard manner and so this procedure ought to serve as a measure of success or failure of the CTA effort.      

 

If the intention is to produce an aversion that reduces predatory attacks upon live prey, then we recommend the following:

 

·   · Nothing in our demonstration suggested that the baits we constructed were flawed in any way and so the details of bait construction described above including clean, fresh, dry hide typical of the target livestock prey securely sutured around the TBZ treated inner ground meat should be followed in future work.  Since the dose of 5.4 g of TBZ well mixed in 250 g of plain ground meat was safe, effective and not apparently detected by the wolves, this should be the standard preparation for baits intended to be consumed by predators the size of these wolves. 

 

·  · Wolves must be subjected to a food deprivation regimen that meets their minimum feeding needs over all, but that includes acute periods of food deprivation from 20-40 hours in duration immediately prior to introduction of baits during all three phases of the protocol.  We consider that a maintenance ration of 12 Kg (26 pounds) of meat per wolf per week distributed in four feedings each of 3 Kg every other day is both adequate to maintain fully grown wolves and sufficient also to induce them to participate in the protocol.  The weekly maintenance diet for wolves proceeding through this protocol should approximate 21% of their body weight as was the case in this demonstration.  The key to this process, however, is to remove all unconsumed food 2-3 hours after each feeding.

 

·   · Barring unforeseen difficulties, presuming that future practitioners can treat wolves separately, and presuming that the intention is solely to establish aversion to live prey and not including an additional effort to produce aversion to carcasses described below, the total amount of time required to complete the protocol might be less than that which we report here.  That is, the food deprivation regimen will require a few days to become established before the protocol itself can begin.  Then, there should be a pre-test of up to 4 days, a single day of treatment and recovery, and a post-test of up to 4 days.  This might total as little as a 10-12 days.

 

·   · Our choice of 60-minute trials reflected our desire to be able to document the full range of largely unpredictable behavior of wolves all maintained together in the same enclosure at the CWC.  In the real world when predator chooses whether or not to pursue prey, seconds are an eternity.  So, forcing hungry wolves to remain in close proximity to a food for which they have an aversion with no alternative food available is a grossly conservative measure of aversion.  If wolves are maintained separately in adequate facilities and moderately deprived, then they should seize and open a bait within a few minutes and very quickly consume the small amount of meat in the interior.  They may then interact with the hide for a time.  Therefore, pre-tests and the treatment might be terminated just as soon as inner meat of baits is consumed and remaining hide is ignored.  Post-testing might require longer to provide assurance that the criteria for successful aversion are met. 

 

·   · It is essential that all of the details of behavior throughout the protocol be documented with a video record.  It is simply not appropriate to attempt to surmise events on the basis of leavings after a trial.  To reduce bias as much as possible, the videotaping may be conducted as unobtrusively as possible.  In this way, review and scoring of events from the video record justify procedure and policy decisions.

 

·   · During the protocol, it is necessary that predators be prevented from consuming any other food for several hours before, during and after eating the baits.   In this way, only the taste and scent of well-constructed baits can be associated with illness during the treatment period.  This is partially accomplished by subjecting wolves to food deprivation as described above but in addition, care should be taken to clean all remnants of earlier feeding so that none might be consumed within three or more hours before or after the baiting period. 

· Captive predators should be proceed through the CTA protocol separately.  If this cannot be done, then baits should be introduced in such a way as to absolutely minimize competition for any single bait that might produce contamination or under-dosing.  The immediate environment should be clean and the substrate clean concrete, gravel or some other substance so that all surrounding contaminants are eliminated.  We strongly suggest that wolves be maintained in a facility large enough for them to move freely toward and away from baits without the introduction of bias that always results from forcing them to interact with baits at close proximity.  If wolves are housed and observed separately, we suggest that interaction with baits take place in facilities that are at least four times the body length of the wolves, including the tail and excluding sleeping areas and that clean water be maintained at all times away from where baits are to be placed.  If wolves have been transferred to their holding facility recently, it is necessary that they demonstrate willingness to consume their maintenance diet and otherwise behave normally before beginning the protocol. 

· Since interest in and interaction with the baits could well be more a function of boredom than predation, we strongly recommend that the wolf environment be greatly enriched with objects and devices designed to interest and occupy the wolves.  An impoverished environment combined with making all stimuli other than food essentially irrelevant to the wolves strongly biases behavioral outcomes, especially during the post-test of this protocol. 

· Although ever wolf in our demonstration required only a single full treatment to form an aversion to baits, this may not always be so.  It is always possible that a wolf might need more than a single full treatment with TBZ before it fulfills the criteria for a successful aversion.  If captive wolves are offered a TBZ bait only once during the treatment period, the first bait offered during the “post-test” should also contain TBZ in case a second treatment is needed.  A wolf that has acquired an aversion to the taste and scent of target livestock should demonstrate a dramatic change in behavior during post-testing compared with its behavior during pre-testing.  That is, the outer surface smell and taste of the bait should be a barrier to a wolf with a satisfactory aversion.  Evidence of this could take the form of ignoring the bait or smell-rejecting the bait.  Wolves might attempt to bury the bait or cover it over.  Since the taste of baits will have become disgusting, if they pick the bait up, they will tend do so only with the incisors dangling the bait from the front of their mouth rather than to hold the bait fully within the mouth.  Some individuals with satisfactory aversion to the bait may nevertheless interact with it by smelling or tasting it.  But their response to the smell and taste of the bait should be pronounced rejection of it and may be accompanied with head shaking, gagging and even retching.  After a few minutes of this, such animals often begin to avoid the bait at a distance and this avoidance should continue during a second post-test. In all of these interactions, if the wolf refuses to interact with the inner contents, the TBZ in the inner meat is irrelevant.  To be assured that this is the case, these wolves can be exposed the next day to another bait without TBZ.  If avoidance at a distance continues then the animal may be considered to have acquired the desired aversion.

· Regardless of whether or not wolves meet the criteria for successful establishment of aversion through this protocol, the taste and scent of the baits used in the protocol must always be close approximations of the taste and scent of livestock on the range where the wolves will be released.

· What to do with “failures”.   Failure in this protocol is much more easily prevented than corrected and the failures discussed below are very unlikely if the protocol is carefully followed.  Possible failures to meet criteria for successful aversion can occur in the following ways: 1. If the animal fails to engage in the sort of rejection behaviors described above and instead, opens the first post-test bait and then fully consumes the inner meat then aversion to both the exterior and the interior of the bait failed in this particular case.  This could be because the TBZ in the interior was insufficient to produce the effect or the wolf detected important differences in the inner and outer scent and taste of the bait during treatment versus the post-test, or the wolf might simply be an unusual individual responding to unusual circumstances.  However, since the wolf consumed the bait as before, it will be treated again with the TBZ in the first “post-test” bait.  After recovering from this second treatment, this animal should meet the post-test criteria for successful aversion on the basis of avoidance of the outer surface as long as the outer taste and scent of treated bait is the same as that for post-test baits. If required avoidance does not develop, refer to #2 below. 2. If an animal is not dissuaded from the outer surface, opens the bait but refuses to fully consume the inner TBZ treated meat then it is possible that the TBZ was detected or, if not detected, aversion to the outer surface failed but aversion to inner meat succeeded.  To resolve between these two possibilities it is necessary to post-test a second time without TBZ in the meat.  If the bait is opened again and plain meat is rejected as before, then aversion to the inner meat succeeded but aversion to outer taste and scent of the bait has failed, almost certainly because of contamination of the outer surface.  If plain meat without TBZ is eaten, then it is likely that somehow the animal detected the TBZ in the meat the first time.  Both of these problems will required an additional treatment to correct but if wolves refuse to consume inner meat with or without TBZ, they cannot be treated again using the same inner meat to establish aversion to the outer surface of the baits.  It might still be possible to establish an aversion to the outer taste and scent of baits and of livestock by changing the bait interior.  Very little is know concerning how to amend a failed aversion, but we suggest that this might be attempted by including some kind of food other than the ground meat originally used.  If it is apparent that the animal initially detected the TBZ in the first meat bait, then the appropriate physiological dose of TBZ may be placed into a larger volume of inner food to help mask the TBZ as long as this volume does not exceed what the wolf is willing to consume in a single meal.  After this, the wolf should be again exposed to baits with the same outer taste and scent.  If this wolf finally avoids the bait from a distance then it may be considered as a success.  If it continues to consume the baits, then it should be scored accordingly and the decision to release or not release this wolf should be made in the light of programmatic needs.  It may be possible to treat this particular animal in the field under more natural settings, should the wolf conflict with livestock.  As with all of the other wolves released after the protocol, the permanent record for wolves that have had difficulties should be maintained and used to interpret future events in the field after their release.

· If they can be induced to voluntarily participate in the protocol, this procedure may be used to treat recently captured animals before they may be released again into habitat with livestock.  The principal difficulty in this case may simply be one of inducing such animals to consume food under the stress of captivity.  This may require that such animals be held in captivity for a longer period of time than might otherwise be planned before their release and it may require that other measures be attempted to induce them to participate.

· Once treated animals have demonstrated aversion to the outer surface of untreated meat baits they may be released and then monitored carefully to determine whether they engage in depredations upon target livestock.  It is during monitoring of wolves in the field that the proper test of the strength and longevity of the aversion effect may take place.  Attempts at measuring these features under captive conditions will inevitably introduce serious bias.

If the intention is to reduce conflicts with livestock by also dissuading predators from scavenging livestock carcasses, then a slight expansion of this protocol could be highly effective: 

· Predators may begin to attack livestock after having scavenged livestock carcasses.  Even if scavenging does not act as a gateway for predation on live prey there are many places where predators are shot or poisoned on sight and so reducing the attractiveness of carcasses may generally reduce the frequency of lethal conflicts with livestock.  To the human nose, decomposing carcasses do not smell like fresh live animals and so non-human predators may also discriminate at some point between carcasses and live prey.  In this way, predators with aversions established with fresh meat baits may nevertheless be attracted to livestock carcasses whose taste and scent sufficiently differ from the baits, with the usual lethal consequences.  The process of putrefaction of whole carcasses is time and temperature-dependent and the products of anaerobic decomposition such as cadaverine, putrescine, and hydrogen sulfide are highly predictable.  Consequently, a fresh bait composed of ground meat tightly wrapped with hide should roughly follow the same course of decomposition events if allowed to remain for a time under warm conditions.  It seems likely that the taste and scent cues of such baits could be reasonable approximations of livestock carcasses and so might be used in an expanded protocol designed to produce aversions that dissuade scavenging as well as predation. This, then, is a microbiological process with important behavioral and practical consequences.  We do not know at this time at what point a bait must decompose so that its taste and scent become altered enough that a predator with an aversion to fresh baits becomes willing to consume the decomposed bait.  Whatever that level of change is will be the limit of influence the aversion protocol will have upon the critical problem of scavenging of livestock remains.  We also do not know how the presence of TBZ might affect the decomposition process.  It is possible that since it has been used as a fungicide TBZ could alter the process in a way that might allow wolves to discriminate between decomposed baits with TBZ and decomposing carcasses.  We also do not know if TBZ would survive in tact through the decomposition process.  To our knowledge, only one attempt to answer this kind of question has been reported.  Nicolaus, et al. 1992 reported that oral estrogen retained its potency as an aversion agent when allowed to remain in eggs through advanced decomposition. We therefore recommend that an attempt be made to determine the safety and efficacy of decomposed TBZ baits using “generic” wolves rather than the much more rare Mexican wolves.  It should be a relatively simple task to repeat the CTA protocol with TBZ meat baits that have decomposed to a point at which they differ markedly from fresh baits and the wolves are still willing to consume them. If it can be shown that decomposed TBZ baits are safe and effective, captive predators with aversion to the exterior of fresh baits may then be offered TBZ-treated baits that have begun to decompose.  If these predators avoid decomposed baits also, then they should systematically be offered TBZ-treated baits with greater and greater decomposition until they finally consume the bait and so are treated.  They should then be post-tested with equally decomposed bait to demonstrate aversion before they may finally be released.  We presume if wolves are consistent in their behavior toward these baits it might be sufficient to simply maintain baits at a known temperature for a known time in order to streamline the second treatment designed to reduce scavenging. 

Establishing Aversions Among Free-ranging Predators

· Nothing requires that the aversion process be carried out among captive predators.  In fact, captive study of predators and behavioral manipulations of captive predators almost always lacks external validity since the conditions of captivity introduce strong and arbitrary biases.  So, once it became known that captive predators such as coyotes and wolves could in principle form an aversion that inhibited attack upon live prey, field trials attempting to establish aversions among free-ranging predators were undertaken (Appendix 1). 

· When done experimentally in the field, ideally several locations populated with predators with a history of depredations upon livestock are chosen randomly as designated treatment areas while others are randomly selected to be controls.  In the treatment locations the process usually requires two phases.  The first phase requires adequate baiting of the free-ranging predator populations in each of the treatment sites and the second requires monitoring of the livestock predation rates attributable to the aversion treated predator populations.  While this is done, predator and livestock prey populations in locations assigned to the control group are merely monitored without aversion treatment. 

· The baiting process in treatment sites requires that the location and movements of predators be either known or approximated as closely as possible.  It requires that a sufficient number of properly constructed baits be placed in an optimum pattern over space and time so that free-ranging predators readily find and consume them.  It requires that this consumption of baits be monitored closely enough so that the fate of each of the baits is known or can be surmised on the basis of the physical evidence.  It requires that baits be recovered and replaced until it is clear that the predator population as a whole has acquired an aversion to the baits and so avoids them at a distance.

· Before, during and after this baiting period, depredation rates by these predator populations upon target livestock is monitored by whatever means is available.  Just the same is monitored in control locations that did not receive treatment.  In general, the difference in depredation rates between these two groups is a measure of the effect of the aversion process.

· Situations in the field are seldom ideal for this sort of experimental design and so what is actually done is a compromise between what must at minimum be done and what can at maximum be done.  Very often the only way to know if an aversion is or is not having an effect is to compare the historical records of depredations at the same locations before aversion treatment with depredation rates after treatment.

· Of course if there is reason to believe that it is safe and effective to do so, TBZ baits placed into the field may be allowed to remain to decompose to some extent in order to confront the problem of scavenging of livestock carcasses. If the observer finds that at some point these are consumed for a while but then are avoided as were the fresh baits before them, this indicates that predators may be dissuaded from scavenging as well.

 

Literature Cited

 

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Bernstein, I.L. 1978. Learned taste aversions in children receiving chemotherapy.  Science. 200:1302-1303.

 

Bernstein, I.L. and Webster, M.M. 1980. Learned taste aversions in humans. Physiol. Behav. 25:363-366.

 

Bernstein, I.L. 1985. Learned food aversions in the progression of cancer and its treatment. Ann. New York Acad. Sci.  443:365-380.

 

Cowan, D.P., Reynolds, J.C. and E.L. Gill. 2000. Reducing predation through conditioned taste aversion. PP 281-299. In L.M. Gosling and W.T. Sutherland (Eds). Behaviour and Conservation. Cambridge University Press, Cambridge.

 

Dimmick, C.R. and L.K. Nicolaus. 1990. Efficiency of conditioned aversion in reducing depredations by crows.  Jour. Appl. Ecol. 27:200-209.

 

Ellins, S.R., Catalono, S.M. and S.A. Schechinger. 1977. Conditioned taste aversion: A field application to coyote predation on sheep. Behav. Biol. 20:91-95.

 

Ellins, S.R., Gustavson, C.R. and J. Garcia. 1978. Conditioned taste aversion in predators: Response to Sterner and Shumake. Behav. Biol. 24:554-556.

 

Gill, E.L. Whiterow, A., and D.P. Cowan. 1999. A comparative assessment of potential conditioned taste aversion agents for vertebrate management.  Appl. Anim. Behav. Sci. 67:229-240.

 

Gustavson, C.R., Garcia, J. Hankins, W.G. and K.W. Rusiniak. 1974. Coyote control by aversive conditioning. Science. 184:581-583.

 

Gustavson, C.R., Kelly, D.J., Sweeney, M. and S. Thomas. 1976. Prey lithium aversions I: Coyotes and wolves. Behav. Biol. 17:61-72.

 

Gustavson, C.R., Kelly, D.J., Sweeney, M. and S. Thomas. 1976. Final report to the Washington Game Department, Olympia, WA.

 

Gustavson, C.R., Jowsey, J.R. and D.N. Milligan. 1982. A three-year evaluation of taste aversion coyote control in Saskatchewan. Jour. Range Manag. 35:57-59.

 

Gustavson, C.R. and Nicolaus, L.K. 1987. Taste aversion conditioning in wolves, coyotes, and other canids: Retrospect and prospect. Pp169-203 In H.Frank (Ed), Man and Wolf: Advances, issues, and problems in captive wolf research.  W.Junk, Boston.

 

Massei, G., and D.P. Cowan. 2002. Strength and persistence of conditioned taste aversion in rats: Evaluation of 11 potential compounds.  Appl. Anim. Behav. Sci. 75:249-260.

 

Massei, G., Lyon, A. and D.P. Cowan. 2003. Potential compounds for inducing conditioned taste aversion in ferrets. New Zealand Journal of Zoology. 30:95-100

 

Massei, G., Lyon, A.J., and D.P. Cowan. 2003. Conditioned taste aversion can reduce egg predation by rats. Jour. Widl. Manage. 66(4):1134-1140.

 

Nicolaus, L.K., Hoffman, T.E., and C.R. Gustavson. 1979. Taste aversion conditioning in raccoons (Procyon lotor). Paper presented to the Portland International Wolf Symposium, August 1979.

 

Nicolaus, L.K., Hoffman, T.E. and C.R. Gustavson. 1982. Taste-aversion conditionig in free-ranging raccoons, Procyon lotor. Northwest Science. 56:156-169.

 

Nicolaus, L.K., Cassel, J.F., Carlson, R.B., and C.R. Gustavson. 1983. Taste-aversion conditioning of crows to control predation on eggs. Science. 220:212-214.

 

Nicolaus, L.K. 1987. Conditioned aversions in a guild of egg predators: Implications for aposematism and prey defense mimicry. Amer. Midl. Natur. 117:405-419.

 

Nicolaus, L. K., and D. Nellis. 1987. The first evaluation of the use of conditioned taste aversion to control predation by mongooses upon eggs. Appl. Anim. Behav. Sci. 17:329-334.

 

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