Serum corticoid levels relative to handling stress in Alaskan moose ALBERTW. FRANZMANN Kenai Moose Research Center, Alaska Department of Fish and Game, Soldotna, Alaska 99669
ARTHURFLYNN Cleveland Metropolitan General Hospital, Case Western Reserve University School of Medicine, Cleveland, Ohio44109 AND
Can. J. Zool. Downloaded from www.nrcresearchpress.com by UNIV OF WISC MADISON on 08/07/13 For personal use only.
PAULD. ARNESON Kenai Moose Research Center, Alaska Department of Fish and Game, Soldotna, Alaska 99669 Received April 30,1975 FRANZMANN, A. W., A. FLYNN, and P. D. ARNESON. 1975.Serum corticoid levels relative to handling stress in Alaskan moose. Can. J. Zool. 53: 1424-1426. Blood serum 1I-hydroxycorticosteroidswere compared to visual evaluation of handling stress in Alaskan moose (Alces alces gigas) at the Kenai Moose Research Center (MRC). Moose were evaluated for handling excitability before and during handling when trapped, and were graded on a scale from 1 (not excited) to 5 (highly excited). There were significant differences in corticosteroid levels (a = 0.1) between each class comparison, except between classes 4and 5,suggesting that this analysis provided a means to classify and compare other blood chemistry values from similarly stressed moose. Other factors may influence the 11-hydroxycorticosteroidlevels, but handling stress had an overwhelming influence. Other methods to evaluate handling stress, such as body temperature, should also be considered, but when not feasible this method may be utilized. FRANZMANN, A. W., A. FLYNNet P. D. ARNESON.1975. Serum corticoid levels relative to handling stress in Alaskan moose. Can. J. Zool. 53: 1424-1426. On a Bvald les 1 I-hydroxycorticosteroi'desdu strum sanguin chez des orignaux d' Alaska (Alces alces gigas) au Centre de Recherches sur les Orignaux (MRC) de Kenai en tenant compte d'une estimation visuelle du stress dti a la manipulation. On a Bvalue I'excitabilite avant et pendant la manipulation au moment de la capture et I'excitabilitt fut cotke de 1 (excitabilitk nulle) a 5 (excitabilite trks elevke). I1 y a des differences significatives dans les niveaux de corticostCroides (a = 0.1) entre les classes, sauf entre les classes 4 et 5, ce qui permet de croire que cette analyse peut servir a classifier et a comparer d'autres facteurs chimiques du sang chez des orignaux ainsi stresses. D'autres facteurs influencent sans doute les niveaux des I I-hydroxycorticosteroides, mais le stress dCi a la manipulation a I'influence predominante. I1 existe d'autres mtthodes d'tvaluation du stress dCi Bla manipulation, par exemple la temperature du corps; on doit en tenir compte, mais la methode proposte ici donne des rtsultats adhuats lorsque les autres ne sont pas utilisables. [Traduit par le journal]
Introduction Evaluating blood chemistry parameters from animals, particularly wild-trapped and immobilized animals, may be confounded by handling stress (Franzmann 1972; Franzmann and Thorne 1970; Gartner et al. 1965, 1969; LeResche et al. 1974; Seal et al. 1972). The need was recognized to standardize the potential effects of handling stress, and an evaluation of individual animal excitability based on heart rate, respiratory rate, and rectal temperature was developed for bighorn sheep (Ovis canadensis) (Franzmann 1972). Each sheep was classified into one of five classes of excitability
(1, not excited; 2, slightly excited; 3, moderately excited; 4, excited; and 5, highly excited). LeResche et al. (1974) suggested that some blood chemicals may change more readily than others in response to stress and provide a means to standardize blood samples. Studies at the Kenai Moose Research Center (MRC), Alaska, have utilized a subjective classification of excitability, adapted from Franzmann (1 972), based upon activity before and during handling of Alaskan moose (Alces alces gigas). This paper reports the use of the classification system and the subsequent analysis of blood serum for 11hydroxycorticosteroids. These steroid hormones
1425
FRANZMANN ET AL.: SERUM CORTICOID LEVELS IN MOOSE
in serum are of clinical significance in the evaluation of adrenocortical function (Catt 1970), and we believed they had potential use as a measured blood value that could potentially reflect handling stress and provide a method to classify and compare samples from similarly stressed animals for other blood chemistry evaluation.
Can. J. Zool. Downloaded from www.nrcresearchpress.com by UNIV OF WISC MADISON on 08/07/13 For personal use only.
Study Area and Methods Moose were sampled at the MRC, which consists of four 2.6 km2enclosures located in the area of the 1947 Kenai burn, 35 km northeast of Soldotna, Alaska. Twenty-two fence-line traps are strategically located (13 within and 9 outside the enclosures) to facilitate the capture and handling of moose (LeResche and Lynch 1973). The traps, when set, were checked daily and when a moose was trapped and immobilized (Franzmann and Arneson 1974); the excitability of the animal before and during immobilization and handling was subjectively evaluated and given a score from one to five (increasing with excitability). Heart rate, respiratory rate, and body temperature were not determined or recorded before the evaluation to remove the effect of potential bias in the evaluation. Blood was collected, centrifuged, and the serum was frozen. In early 1974, samples were analyzed from each month sampled during 1973 to compare samples through all seasons. An attempt was made to obtain samples from each of the five excitability classes. No samples were available for May, but 11 were analyzed from the January sample and 10 were analyzed from each of the remaining months of the year for a total sample of 111. No selection was made based upon sex or age; however, a representative sample of the sex and age structure of the population was obtained. The serum samples were sent to the Department of Surgery, Cleveland Metropolitan General Hospital, Case Western Reserve University School of Medicine, Cleveland, Ohio, to be fluorometrically analyzed for plasma 11-hydroxycorticosteroidsby the rapid procedure for clinical screening as outlined by Mejer and Blanchard (1973).
Results Table 1 lists sample size, mean, and standard deviation for 11-hydroxycorticosteroidsfor each moose excitability class. Class 1 (not excited) had 7 samples with a mean of 26.6 pg/100 ml, class 2 (slightly excited) had 51 samples with a mean of 48.3 pg/100 ml, class 3 (moderately excited) had 28 samples with a mean of 61.9 pg/ 100 ml, class 4 (excited) had 10 samples with a mean of 104.2 pg/lOOml, and class 5 (highly excited) had 15 samples with a mean of 114.5 pg/ 100 ml. An analysis of variance was computed with the hypothesis that there were no differences
TABLE 1 Average 11-hydroxycorticosteroid levels (pg/100 ml) in moose blood serum for five excitability classes Excitability class
Sample size Mean SD
1
2
3
4
5
7 26.6 9.06
51 48.3 13.71
28 61.9 11.90
10 104.2 14.26
15 114.5 20.72
between excitability classes. This hypothesis was rejected at a 1% level of significance. Scheffe's S-value test was computed to determine where the differences occurred, and we found all classes were significantly different from each other except classes 4 and 5.
Discussion With the array of excitability-class means distributed in an increasing manner corresponding to the increase in excitability, and with significant differences between each excitability class, except between classes 4 and 5, we concluded that serum analysis of 1l-hydroxycorticosteroids provided a means to identify samples of similarly stressed moose for other blood chemistry analyses. It also was evident that a subjective evaluation of excitability stress before and during handling of captured moose was a valid procedure since the distinct corticosteroid class averages indicated that experienced observers consistently evaluated the excitability stress of moose. Since the two most highly rated excitability classes (4 and 5) were not significantly different, it may be feasible to use four rather than five excitability classes for moose. This would permit less dilution of sample size for standardization of sample. A suggested classification based on these data would be as follows: class 1, all values below 35 pg/100 ml; class 2, all values from 35 to 55 pg/100 ml; class 3, all values from 55 to 75 pg/100 ml; and class 4, all values from 75 pg/100 ml and above. We believed that seasonal, as well as age and sex, differences would influence findings, therefore, we made sure the data contained a generous number of samples comprising all seasons, sexes, and ages. The results indicate that the
Can. J. Zool. Downloaded from www.nrcresearchpress.com by UNIV OF WISC MADISON on 08/07/13 For personal use only.
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CAN. J. ZOOL. VOL. 53, 1975
influence of these other factors on corticosteroid CATT,K. J. 1970. Adrenal cortex.lancet l970(i): 1275. A. W. 1972. Environmental sources of varialevels was minor compared to handling stress. FRANZMANN, tion of bighorn sheep physiologic values. J. Wildl. ManIn this study, the overwhelming influence of age. 36: 924-932. handling stress was represented by I l-hydroxy- FRANZMANN, A. W., and P. D. ARNESON. 1974. Immobilicorticosteroid levels. zation of Alaskan moose. J. Zoo Anim. Med. 5: 26-32. A. W., andE. T. THORNE.1970. Physiologic We do not suggest that 1 l-hydroxycortico- FRANZMANN, values in wild bighorn sheep (Ovis canadensis canasteroid analysis be the sole criteria for evaluating densis) at capture, after handling, and after captivity. J. handling stress, but that it be considered as an Am. Vet. Med. Assoc. 157: 647-650. additional tool to supplement this evaluation. GARTNER, R. J. W., L. L. CALLOW,C. K. GRAZIEN, and P. PEPPER.1969. Variations in the concentration of blood Body temperature has provided a means to do constituents in relation to handling cattle. Res. Vet. Sci. this (Franzmann 1972), but may not be feasible 10: 7-12. in all instances. Without body temperature, a GARTNER, R. J . W., J . W. RYLEY,and A. W. BEATTIE. handling stress evaluation can be made by this 1965. The influence of degree of excitation on certain blood constituents in beef cattle. Aust. J. Exp. Biol. procedure. Other blood chemistries that poMed. Sci. 43: 713-724. tentially could reflect handling stress should be LERESCHE, R. E., and G. W. LYNCH.1973. A trap for investigated. free-ranging moose. J . Wildl. Manage. 37: 87-89.
Acknowledgments The Kenai Moose Research Center is a cooperative project of the Alaska Department of Fish and Game and the U.S. Fish and Wildlife Service, Kenai National Moose Range. This work was supported, in part, by Federal Aid in Wildlife Restoration Projects W-17-R. We thank J. L. Oldemeyer for assistance with statistical analysis and for critical review of the manuscript. K. B. Schneider and D. M. McKnight also reviewed the manuscript and provided helpful suggestions.
LERESCHE,R. E., U. S . SEAL,P. D. KARNS,and A. W. 1974. A review of blood chemistry of FRANZMANN. moose and other Cervidae, with emphasis on nutritional assessment. Nat. Can. (Quebec), 101: 263-290. MEJER,L. E . , and R. C. BLANCHARD. 1973. Fluorometric determination of plasma 11-hydroxycorticosteroids.1 . Rapid procedure for clinical screening. Clin. Chem. 19: 710-717. SEAL,U. S., J. J. OZOGA,A. W. ERICKSON,and L. J. VERME.1972. Effects of immobilization on blood analysis of white-tailed deer. J. Wildl. Manage. 36: 10341040.
ARTHURFLYNN Cleveland Metropolitan General Hospital, Case Western Reserve University School of Medicine, Cleveland, Ohio44109 AND
Can. J. Zool. Downloaded from www.nrcresearchpress.com by UNIV OF WISC MADISON on 08/07/13 For personal use only.
PAULD. ARNESON Kenai Moose Research Center, Alaska Department of Fish and Game, Soldotna, Alaska 99669 Received April 30,1975 FRANZMANN, A. W., A. FLYNN, and P. D. ARNESON. 1975.Serum corticoid levels relative to handling stress in Alaskan moose. Can. J. Zool. 53: 1424-1426. Blood serum 1I-hydroxycorticosteroidswere compared to visual evaluation of handling stress in Alaskan moose (Alces alces gigas) at the Kenai Moose Research Center (MRC). Moose were evaluated for handling excitability before and during handling when trapped, and were graded on a scale from 1 (not excited) to 5 (highly excited). There were significant differences in corticosteroid levels (a = 0.1) between each class comparison, except between classes 4and 5,suggesting that this analysis provided a means to classify and compare other blood chemistry values from similarly stressed moose. Other factors may influence the 11-hydroxycorticosteroidlevels, but handling stress had an overwhelming influence. Other methods to evaluate handling stress, such as body temperature, should also be considered, but when not feasible this method may be utilized. FRANZMANN, A. W., A. FLYNNet P. D. ARNESON.1975. Serum corticoid levels relative to handling stress in Alaskan moose. Can. J. Zool. 53: 1424-1426. On a Bvald les 1 I-hydroxycorticosteroi'desdu strum sanguin chez des orignaux d' Alaska (Alces alces gigas) au Centre de Recherches sur les Orignaux (MRC) de Kenai en tenant compte d'une estimation visuelle du stress dti a la manipulation. On a Bvalue I'excitabilite avant et pendant la manipulation au moment de la capture et I'excitabilitt fut cotke de 1 (excitabilitk nulle) a 5 (excitabilite trks elevke). I1 y a des differences significatives dans les niveaux de corticostCroides (a = 0.1) entre les classes, sauf entre les classes 4 et 5, ce qui permet de croire que cette analyse peut servir a classifier et a comparer d'autres facteurs chimiques du sang chez des orignaux ainsi stresses. D'autres facteurs influencent sans doute les niveaux des I I-hydroxycorticosteroides, mais le stress dCi a la manipulation a I'influence predominante. I1 existe d'autres mtthodes d'tvaluation du stress dCi Bla manipulation, par exemple la temperature du corps; on doit en tenir compte, mais la methode proposte ici donne des rtsultats adhuats lorsque les autres ne sont pas utilisables. [Traduit par le journal]
Introduction Evaluating blood chemistry parameters from animals, particularly wild-trapped and immobilized animals, may be confounded by handling stress (Franzmann 1972; Franzmann and Thorne 1970; Gartner et al. 1965, 1969; LeResche et al. 1974; Seal et al. 1972). The need was recognized to standardize the potential effects of handling stress, and an evaluation of individual animal excitability based on heart rate, respiratory rate, and rectal temperature was developed for bighorn sheep (Ovis canadensis) (Franzmann 1972). Each sheep was classified into one of five classes of excitability
(1, not excited; 2, slightly excited; 3, moderately excited; 4, excited; and 5, highly excited). LeResche et al. (1974) suggested that some blood chemicals may change more readily than others in response to stress and provide a means to standardize blood samples. Studies at the Kenai Moose Research Center (MRC), Alaska, have utilized a subjective classification of excitability, adapted from Franzmann (1 972), based upon activity before and during handling of Alaskan moose (Alces alces gigas). This paper reports the use of the classification system and the subsequent analysis of blood serum for 11hydroxycorticosteroids. These steroid hormones
1425
FRANZMANN ET AL.: SERUM CORTICOID LEVELS IN MOOSE
in serum are of clinical significance in the evaluation of adrenocortical function (Catt 1970), and we believed they had potential use as a measured blood value that could potentially reflect handling stress and provide a method to classify and compare samples from similarly stressed animals for other blood chemistry evaluation.
Can. J. Zool. Downloaded from www.nrcresearchpress.com by UNIV OF WISC MADISON on 08/07/13 For personal use only.
Study Area and Methods Moose were sampled at the MRC, which consists of four 2.6 km2enclosures located in the area of the 1947 Kenai burn, 35 km northeast of Soldotna, Alaska. Twenty-two fence-line traps are strategically located (13 within and 9 outside the enclosures) to facilitate the capture and handling of moose (LeResche and Lynch 1973). The traps, when set, were checked daily and when a moose was trapped and immobilized (Franzmann and Arneson 1974); the excitability of the animal before and during immobilization and handling was subjectively evaluated and given a score from one to five (increasing with excitability). Heart rate, respiratory rate, and body temperature were not determined or recorded before the evaluation to remove the effect of potential bias in the evaluation. Blood was collected, centrifuged, and the serum was frozen. In early 1974, samples were analyzed from each month sampled during 1973 to compare samples through all seasons. An attempt was made to obtain samples from each of the five excitability classes. No samples were available for May, but 11 were analyzed from the January sample and 10 were analyzed from each of the remaining months of the year for a total sample of 111. No selection was made based upon sex or age; however, a representative sample of the sex and age structure of the population was obtained. The serum samples were sent to the Department of Surgery, Cleveland Metropolitan General Hospital, Case Western Reserve University School of Medicine, Cleveland, Ohio, to be fluorometrically analyzed for plasma 11-hydroxycorticosteroidsby the rapid procedure for clinical screening as outlined by Mejer and Blanchard (1973).
Results Table 1 lists sample size, mean, and standard deviation for 11-hydroxycorticosteroidsfor each moose excitability class. Class 1 (not excited) had 7 samples with a mean of 26.6 pg/100 ml, class 2 (slightly excited) had 51 samples with a mean of 48.3 pg/100 ml, class 3 (moderately excited) had 28 samples with a mean of 61.9 pg/ 100 ml, class 4 (excited) had 10 samples with a mean of 104.2 pg/lOOml, and class 5 (highly excited) had 15 samples with a mean of 114.5 pg/ 100 ml. An analysis of variance was computed with the hypothesis that there were no differences
TABLE 1 Average 11-hydroxycorticosteroid levels (pg/100 ml) in moose blood serum for five excitability classes Excitability class
Sample size Mean SD
1
2
3
4
5
7 26.6 9.06
51 48.3 13.71
28 61.9 11.90
10 104.2 14.26
15 114.5 20.72
between excitability classes. This hypothesis was rejected at a 1% level of significance. Scheffe's S-value test was computed to determine where the differences occurred, and we found all classes were significantly different from each other except classes 4 and 5.
Discussion With the array of excitability-class means distributed in an increasing manner corresponding to the increase in excitability, and with significant differences between each excitability class, except between classes 4 and 5, we concluded that serum analysis of 1l-hydroxycorticosteroids provided a means to identify samples of similarly stressed moose for other blood chemistry analyses. It also was evident that a subjective evaluation of excitability stress before and during handling of captured moose was a valid procedure since the distinct corticosteroid class averages indicated that experienced observers consistently evaluated the excitability stress of moose. Since the two most highly rated excitability classes (4 and 5) were not significantly different, it may be feasible to use four rather than five excitability classes for moose. This would permit less dilution of sample size for standardization of sample. A suggested classification based on these data would be as follows: class 1, all values below 35 pg/100 ml; class 2, all values from 35 to 55 pg/100 ml; class 3, all values from 55 to 75 pg/100 ml; and class 4, all values from 75 pg/100 ml and above. We believed that seasonal, as well as age and sex, differences would influence findings, therefore, we made sure the data contained a generous number of samples comprising all seasons, sexes, and ages. The results indicate that the
Can. J. Zool. Downloaded from www.nrcresearchpress.com by UNIV OF WISC MADISON on 08/07/13 For personal use only.
1426
CAN. J. ZOOL. VOL. 53, 1975
influence of these other factors on corticosteroid CATT,K. J. 1970. Adrenal cortex.lancet l970(i): 1275. A. W. 1972. Environmental sources of varialevels was minor compared to handling stress. FRANZMANN, tion of bighorn sheep physiologic values. J. Wildl. ManIn this study, the overwhelming influence of age. 36: 924-932. handling stress was represented by I l-hydroxy- FRANZMANN, A. W., and P. D. ARNESON. 1974. Immobilicorticosteroid levels. zation of Alaskan moose. J. Zoo Anim. Med. 5: 26-32. A. W., andE. T. THORNE.1970. Physiologic We do not suggest that 1 l-hydroxycortico- FRANZMANN, values in wild bighorn sheep (Ovis canadensis canasteroid analysis be the sole criteria for evaluating densis) at capture, after handling, and after captivity. J. handling stress, but that it be considered as an Am. Vet. Med. Assoc. 157: 647-650. additional tool to supplement this evaluation. GARTNER, R. J. W., L. L. CALLOW,C. K. GRAZIEN, and P. PEPPER.1969. Variations in the concentration of blood Body temperature has provided a means to do constituents in relation to handling cattle. Res. Vet. Sci. this (Franzmann 1972), but may not be feasible 10: 7-12. in all instances. Without body temperature, a GARTNER, R. J . W., J . W. RYLEY,and A. W. BEATTIE. handling stress evaluation can be made by this 1965. The influence of degree of excitation on certain blood constituents in beef cattle. Aust. J. Exp. Biol. procedure. Other blood chemistries that poMed. Sci. 43: 713-724. tentially could reflect handling stress should be LERESCHE, R. E., and G. W. LYNCH.1973. A trap for investigated. free-ranging moose. J . Wildl. Manage. 37: 87-89.
Acknowledgments The Kenai Moose Research Center is a cooperative project of the Alaska Department of Fish and Game and the U.S. Fish and Wildlife Service, Kenai National Moose Range. This work was supported, in part, by Federal Aid in Wildlife Restoration Projects W-17-R. We thank J. L. Oldemeyer for assistance with statistical analysis and for critical review of the manuscript. K. B. Schneider and D. M. McKnight also reviewed the manuscript and provided helpful suggestions.
LERESCHE,R. E., U. S . SEAL,P. D. KARNS,and A. W. 1974. A review of blood chemistry of FRANZMANN. moose and other Cervidae, with emphasis on nutritional assessment. Nat. Can. (Quebec), 101: 263-290. MEJER,L. E . , and R. C. BLANCHARD. 1973. Fluorometric determination of plasma 11-hydroxycorticosteroids.1 . Rapid procedure for clinical screening. Clin. Chem. 19: 710-717. SEAL,U. S., J. J. OZOGA,A. W. ERICKSON,and L. J. VERME.1972. Effects of immobilization on blood analysis of white-tailed deer. J. Wildl. Manage. 36: 10341040.
