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Yield of venom from the Osage copperhead, Agkistrodon contortrix phaeogaster
T°sic°a, VoL 20, No. 2, pp. 525 521. 1982 Printed in Great Britain.
0041-0101/82/020525-03 503 .00/0 p 1982 Pergam°a Prey Ltd.
YIELD OF VENOM FROM THE OSAGE COPPERHEAD, AGKISTRODON CONTORTRIX PHAEOGASTER CYNTHIA A. DE W Tr s Physiology Department of and Cell Biology, University of Kansas, Lawrence, KS 66045, U .S.A . (Accepted jor publication 8 September 1981)
C. A .
DE Wrr. Yield of venom from the Osage copperhead, Agkistrodon contortrix phaeogaster. Toxicon 20, 525-527, 1982 .-Venom was extracted from eighteen Osage copperheads, Agkistrodon contortrix phaeogaster, by electrical stimulation . Mean venom yield was 43 .4 mg/sna~e and mean
body length was 57.5 cm (snout-venu Solid content was 28 .3 mg dry venom/100 ml fresh venom. These yields were comparable to those obtained for three other subspecies .
there are five recognized subspecies of copperheads (Gloyd and Conant, 1943 ; Gloyd, 1969), venom yields are available only for Agkistrodon contortriz contortrix, A. c. laticinctus and A. c. mokasen. This paper reports venom yields for a fourth subspecies, A. c. phaeogaster. Eighteen adult Agkistrodon contortrix phaeogaster were collected at several sites in Douglas and western Johnson counties in Kansas during the spring and fall of 1979 and spring of 1980. The method ofcaptureentailed turning over rocks on sunny slopes to Imcover the snakes, then gently picking the snakes up with tongs and placing them in a bag. When gripped by the tongs, some snakes struck and thus lost some venom. After capture, the snakes were kept in cages at temperatures of21-24°C and were allowed at least two weeks to recover any venom lost during capture. Venom was then extracted by electrical stimulation (Johnson, 1938 ; Glenn,1972). After venom extraction the snakes were released, except for three snakes caught in the fall of 1979. They were fed mice and newborn rats every two to three weeks and were extracted three times during their captivity. Venom extraction was performed on four different occasions. Venom was collected because this subspecies'venom is not commercially available. Because a study ofvenom yield was not the original reason for extracting the snakes,neither the volume ofthe venom nor the length ofthe snakes was measured the first three times. However, on the fourth extraction, a large number of snakes were available and these parameters were measured . On each occasion, after all snakes were extracted, the pooled venom was centrifuged at 20,000 p for 30 min at 0°C to remove cellular debris. It was then frozen and lyophilized overnight. After lyophilization the venom was weighed, transferred to a dark vial and stored at -20°C. After weighing, the venom from all four extractions was placed in the same vial ALTHOUGH
" Address as of 1 September 1981 : Heart Section, Department of Medicine, General Hospital, 5-214 Ol Malmö, Sweden. 525
526
Short Communications
TABLE I. MEAN LENGTH AND VENOM YIELD FROM Agkistrodon contortrix phaeogasttr (18 SNAKFS) OBTAINED BY ELECTRICAL STIMULATION
Dates of extraction "
Number of snakes extracted
4 May, 1979 2 November, 1979 6 March, 1980 5 May, 1980 Total
Snake identification numbers
6 3 3 12 24
1-6 7-9 7-9 7-18 ~
Total dried venom (mg)
Total volume of fresh venom (ml)
Mean snout-vent length (cm)t
Mean venom yield (mg/snake)
270 105 171 495 1041
1.75
57 .5±2.5
45 34.9 57 41 .3 43 .4
"For 4 May,1979 and 2 November,1979 the snakes were caught approximately two weeks before extraction . The snakes extracted on 6 March 1980 were the same as those extracted 2November and the snakes extracted on 5 May included these same three snakes plus nine wild snakes caught approximately two weeks before. tMean t S.E. TABLE 2. MEAN LENGTHS AND VENOM YIELDS FROM FOUR SUBSFECB"S OF Agkistrodon contortrix
A. c. A. c. A. c. A. c.
Subspecies
Mean snout-vent length (cm)
phaeogasttr contwtrix mokastn laticinctus
57 .5 t 2.5 76t 73.5§ 66II
Mean venom yield mg/snake ml/snake mg/cm 43 .4 57.5$ 57.8§ SOt
0.146 0.20§ -
0.72 0.76 0.79 0.76
Percent solids" (mgJ100m1) 28.3 33 .9§ -
"Percent solids, in mg of dry venom per IOOmI of fresh venom. tRussell (1967) $Russell (1980). §Minion (1953) . ~~Conant (1975).
Results from the four extractions are given in Table 1. The average values for several other subspecies of copperheads are given in Table 2. Differences in venom yields in wild versus captive snakes can be seen in Table 1 . The snakes extracted on 6 March were first extracted on 2 November shortly after their capture. After being kept for four months their venom yield increased. There are many other parameters that can affect the yield of venom, including the sex of the snake, its general health, the time of year, whether it has eaten recently, etc. It is düiïcult to take all parameters into account when measuring venom yields. In this study the sex ofthe snake was not noted as all thevenom was pooled, and the general health ofthe wild snakes could not be determined during such a short captivity. Nevertheless, the yields in this study fall within the range of values reported for the species as a whole. Acknowledgcmcnts-I thank HANK GUARISCO for technical assistance and ANNIKA NltssoN, MARK KozueowsKl and HARVEY LILLYWHITE for the critical review of this manuscript. Tbis research was supported by graduate teaching funds provided by the University of Kansas. REFERENCES
CONANT, R. (1975) A Field Guide to Reptiles and Amphibians ojEastern and Central North America. Boston : Houghton Mi®in Co. GLENN, J. L. (1972) Yield of venom obtained from Crotales atrox by electrical stimulation. Toxicon 10, 575. LLOYD, H. K. (1969) Two additional subspecies of North American crotalid snakes, genus Agkistrodon. Proc. biol. Soc. Wash 82, 219.
Short Communications
52 7
Gt .om, H . K. and CONANT, R . (1943) A synopsis of the American forms of Agkistrodon. Bull. Chicago Acad. Sci. 7, 147 . JOHNSON, C . M. (1938) A new method for stripping venomous snakes . Am . J. trop. Med. 18, 385 . Mixrox, S. A. (1953) Variation in venom samples from copperheads (Agkistrodon contortrix mokeson) and timber rattlers (Crotales horridus horridus). Copeia 1953, 212. Russet, F. E . (1967) Pharmacology of animal venoms . Clip. Pharmac. Then. S, 849. ~RussEt.t ., F. E . (1980) Snake Venom Poisoning. Philadelphia : J. B . Lippinwtt Co.
0041-0101/82/020525-03 503 .00/0 p 1982 Pergam°a Prey Ltd.
YIELD OF VENOM FROM THE OSAGE COPPERHEAD, AGKISTRODON CONTORTRIX PHAEOGASTER CYNTHIA A. DE W Tr s Physiology Department of and Cell Biology, University of Kansas, Lawrence, KS 66045, U .S.A . (Accepted jor publication 8 September 1981)
C. A .
DE Wrr. Yield of venom from the Osage copperhead, Agkistrodon contortrix phaeogaster. Toxicon 20, 525-527, 1982 .-Venom was extracted from eighteen Osage copperheads, Agkistrodon contortrix phaeogaster, by electrical stimulation . Mean venom yield was 43 .4 mg/sna~e and mean
body length was 57.5 cm (snout-venu Solid content was 28 .3 mg dry venom/100 ml fresh venom. These yields were comparable to those obtained for three other subspecies .
there are five recognized subspecies of copperheads (Gloyd and Conant, 1943 ; Gloyd, 1969), venom yields are available only for Agkistrodon contortriz contortrix, A. c. laticinctus and A. c. mokasen. This paper reports venom yields for a fourth subspecies, A. c. phaeogaster. Eighteen adult Agkistrodon contortrix phaeogaster were collected at several sites in Douglas and western Johnson counties in Kansas during the spring and fall of 1979 and spring of 1980. The method ofcaptureentailed turning over rocks on sunny slopes to Imcover the snakes, then gently picking the snakes up with tongs and placing them in a bag. When gripped by the tongs, some snakes struck and thus lost some venom. After capture, the snakes were kept in cages at temperatures of21-24°C and were allowed at least two weeks to recover any venom lost during capture. Venom was then extracted by electrical stimulation (Johnson, 1938 ; Glenn,1972). After venom extraction the snakes were released, except for three snakes caught in the fall of 1979. They were fed mice and newborn rats every two to three weeks and were extracted three times during their captivity. Venom extraction was performed on four different occasions. Venom was collected because this subspecies'venom is not commercially available. Because a study ofvenom yield was not the original reason for extracting the snakes,neither the volume ofthe venom nor the length ofthe snakes was measured the first three times. However, on the fourth extraction, a large number of snakes were available and these parameters were measured . On each occasion, after all snakes were extracted, the pooled venom was centrifuged at 20,000 p for 30 min at 0°C to remove cellular debris. It was then frozen and lyophilized overnight. After lyophilization the venom was weighed, transferred to a dark vial and stored at -20°C. After weighing, the venom from all four extractions was placed in the same vial ALTHOUGH
" Address as of 1 September 1981 : Heart Section, Department of Medicine, General Hospital, 5-214 Ol Malmö, Sweden. 525
526
Short Communications
TABLE I. MEAN LENGTH AND VENOM YIELD FROM Agkistrodon contortrix phaeogasttr (18 SNAKFS) OBTAINED BY ELECTRICAL STIMULATION
Dates of extraction "
Number of snakes extracted
4 May, 1979 2 November, 1979 6 March, 1980 5 May, 1980 Total
Snake identification numbers
6 3 3 12 24
1-6 7-9 7-9 7-18 ~
Total dried venom (mg)
Total volume of fresh venom (ml)
Mean snout-vent length (cm)t
Mean venom yield (mg/snake)
270 105 171 495 1041
1.75
57 .5±2.5
45 34.9 57 41 .3 43 .4
"For 4 May,1979 and 2 November,1979 the snakes were caught approximately two weeks before extraction . The snakes extracted on 6 March 1980 were the same as those extracted 2November and the snakes extracted on 5 May included these same three snakes plus nine wild snakes caught approximately two weeks before. tMean t S.E. TABLE 2. MEAN LENGTHS AND VENOM YIELDS FROM FOUR SUBSFECB"S OF Agkistrodon contortrix
A. c. A. c. A. c. A. c.
Subspecies
Mean snout-vent length (cm)
phaeogasttr contwtrix mokastn laticinctus
57 .5 t 2.5 76t 73.5§ 66II
Mean venom yield mg/snake ml/snake mg/cm 43 .4 57.5$ 57.8§ SOt
0.146 0.20§ -
0.72 0.76 0.79 0.76
Percent solids" (mgJ100m1) 28.3 33 .9§ -
"Percent solids, in mg of dry venom per IOOmI of fresh venom. tRussell (1967) $Russell (1980). §Minion (1953) . ~~Conant (1975).
Results from the four extractions are given in Table 1. The average values for several other subspecies of copperheads are given in Table 2. Differences in venom yields in wild versus captive snakes can be seen in Table 1 . The snakes extracted on 6 March were first extracted on 2 November shortly after their capture. After being kept for four months their venom yield increased. There are many other parameters that can affect the yield of venom, including the sex of the snake, its general health, the time of year, whether it has eaten recently, etc. It is düiïcult to take all parameters into account when measuring venom yields. In this study the sex ofthe snake was not noted as all thevenom was pooled, and the general health ofthe wild snakes could not be determined during such a short captivity. Nevertheless, the yields in this study fall within the range of values reported for the species as a whole. Acknowledgcmcnts-I thank HANK GUARISCO for technical assistance and ANNIKA NltssoN, MARK KozueowsKl and HARVEY LILLYWHITE for the critical review of this manuscript. Tbis research was supported by graduate teaching funds provided by the University of Kansas. REFERENCES
CONANT, R. (1975) A Field Guide to Reptiles and Amphibians ojEastern and Central North America. Boston : Houghton Mi®in Co. GLENN, J. L. (1972) Yield of venom obtained from Crotales atrox by electrical stimulation. Toxicon 10, 575. LLOYD, H. K. (1969) Two additional subspecies of North American crotalid snakes, genus Agkistrodon. Proc. biol. Soc. Wash 82, 219.
Short Communications
52 7
Gt .om, H . K. and CONANT, R . (1943) A synopsis of the American forms of Agkistrodon. Bull. Chicago Acad. Sci. 7, 147 . JOHNSON, C . M. (1938) A new method for stripping venomous snakes . Am . J. trop. Med. 18, 385 . Mixrox, S. A. (1953) Variation in venom samples from copperheads (Agkistrodon contortrix mokeson) and timber rattlers (Crotales horridus horridus). Copeia 1953, 212. Russet, F. E . (1967) Pharmacology of animal venoms . Clip. Pharmac. Then. S, 849. ~RussEt.t ., F. E . (1980) Snake Venom Poisoning. Philadelphia : J. B . Lippinwtt Co.