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The toxicity of T-2 toxin and diacetoxyscirpenol in combination for broiler chickens
oOlS-62~/gl/wOlSS~SO2.~/O Copyright 0 1981 Pcrgamon Press Ltd
Fd Cosmrr. Toricol. Vol. 19, pp. I85 10 188. 1981 Printed in Great Britain. All rights reserved
THE TOXICITY OF T-2 TOXIN AND DIACETOXYSCIRPENOL IN COMBINATION FOR BROILER CHICKENS F.J. HOERR and W.W. CARLTON Department
of Veterinary
Microbiology, Pathology and Purdue University, Lafayette,
Public Health, IN
47907,
School USA
of Veterinary
Medicine,
and B. YAGEN Department
01 Natural
Products,
School Jerusalem,
(Received
of Pharmacy, Israel
2SAugust
The Hebrew
University,
1980)
Abstract-T-2 toxin and diacetoxyscirpenol,two naturally occurring trichothecenemycotoxins,were dissolved in dimethylsulphoxide:saline (1:9, v/v) and administered separately and in combinations to 7-day-old male broiler chickens. The 72-hr single oral dose LD,, for T-2 toxin was 4.0mg/kg body .weight and for diacetoxyscirpenol. it was 5.0 mg/kg. T-2 toxin administered as 14 daily oral doses had an LDse of 2,90mg/kg, and diacetoxyscirpenol had an LD,, of 4.15 mg/kg. Combinations of the toxins caused additive lethal effects in both the single- and multiple-dose tests.
INTRODUCTION The 12,13-epoxy-trichothecenes are a group of at least 40 structurally related toxic fungal metabolites, most of which are produced by Fusarium species (Mirocha, 1979). Four of these mycotoxins, T-2 toxin, diacetoxyscirpenol, deoxynivalenol and nivalenol, have been identified as toxic components of naturally contaminated feedstuffs. Of these four toxins, two have been found in a single feed sample on at least two occasions (Ghosal, Biswas, Srivastava, Chakrabarti & Basu Chaudhary, 1978; Mirocha, Schauerhamer, Christensen & Kommedahl, 1979),and several species of Fusarium can produce combinations of these and other trichothecenes when cultured under laboratory conditions (Ciegler, 1978; Ueno, Ishii, Sawano, Ohtsubo. Matsuda, Tanaka, Kurata & Ichinoe, 1977). Synergistic lethal toxicity of three metabelites of Penicillium uiridicatum (ochratoxin A, citrinin, and penicillic acid) was demonstrated in female mice (&sing, Lillehoj, Detroy 8~ Miller, 1976). This paper describes the toxicity of T-2 toxin and diacetoxyscirpenol, given separately and in combinations as single and multiple doses, in male broiler chickens. EXPERIMENTAL
Chickens, Male broiler chicks of the Hubbard x Hubbard strain were obtained when one day old from a commercial hatchery (Fairview Farms, Remington, IN). The parent flock was seronegative to Myc@ plasma gallisepticum and M. synouiae, and the chicks received Marek’s disease vaccine prior to shipment. They were reared in wire-floored brooding cages measuring 60 x 90 x 20cm, with a brooding temperature of 3540°C a room temperature of 22°C continuous illumination, and forced air ventilation. A maximum of 50 birds were housed in each cage until 185
they were 14 days old when the maximum density was reduced to 32 birds per cage. Non-medicated commercial starter mash for broiler chicks and fresh tap water were available ud lib. The chicks were acclimatized and observed daily but not handled until they were 7 days old, when the body weight was about 100g. Sequentially numbered aluminium wingbands were randomized, affixed to the wing web, and the birds were placed in dose groups according to the wingband number sequence. Toxins and solvent. T-2 toxin was prepared by B. Yagen and diacetoxyscirpenol was obtained from Makor Chemical, Ltd. (Jerusalem, Israel). Each toxin was dissolved in one part reagent-grade dimethylsulphoxide and then diluted to a final volume with nine parts of physiological saline to make stock solutions of appropriate concentrations for administration to the high-dose groups. These stock toxin solutions were diluted to the desired concentrations with a stock solvent solution of dimethylsulphoxide:saline (1:9 v/v). The solvent exposure was 10 ml/kg body weight regardlessof the toxin dose. For the multiple dose study, the toxic solutions were prepared daily and administered within 3 hr of preparation. Toxin administration. Single oral dose 72-hr LDs, determinations were made for each toxin given separately and in dosage combinations presented in Table 1. The toxins were administered into the crop using a l-ml plastic tuberculin syringe and a curved 75cm dosing needle. Feed was withheld 4 hr prior to dosing The LD,e was calculated by the method of Litchfield & Wilcoxon (1949) and the joint potency was defined by the isobologram method of Hewlett (1969) for two drugs of separate activity. Ten birds were in each dose group including a solvent-only group. Multiple oral dose LD,, determinations were based on 14 consecutive daily doses administered to dose groups described in Table 1. The method of
186
F. J. HOERR. Table
W. W. CARLTON
and
1. T-2 toxin
and diacetoxyscirpenol single and Zl-day multiple
73hr
combinations oral toxicity
T-2 toxin: diacetoxyscirpenol dosages
(mg/kg body weight)
4.5 : I .75 4.0: 1.5 3.5:1.25 3.0: 1.0
no. in group)
4.0:0 (2.74-5.84:O)
3.25: 1.14 (3.01-3.51 : 1.06-1.22)
S/IO 4110 4/10 l/l0 l/l0
2.2612.26 (I ,74-2.94: 1.74-2.94)
IO/IO IO/IO S/IO S/IO
0:60 0:5.0 0:4.0 0:3.0 0:2.0 0: 1.0
1.03 : 3.05 (0.82-1~28:244-3.81)
4110 5110 5110 o/to O/IO O/IO Multiple-dose 5110 4110 O/IO O/IO
2.50: I.0 2.25 ~0.75 2GO:0~50 1.75:@25
518 3/8
1.75: 1.50: 1.25: 1.00:
1.75 1.50 1.25 I.00
5/8 318
1.0:2.50 0.75 12.25 0~50:2GG 0.25: 1.75
518
0:3.5 0:3,0 0:2.5
toxin administration was similar to that in the singledose study except that feed was withheld 2 to 3 hr prior to dose administration. For individual toxin administration and solvent-only controls, there were ten birds per dose group. Eight birds were in each group that received the combined toxins. RESULTS toxicity
The toxins administered alone and in combinations usually caused death within 24 hr of dosing. AfTected
intervals
test
IO/10 S/IO S/IO 4/10
1.75:4.5 1.5:4.0 1.25:3.5 1.0:3.0
Single-dose
for
toxin combinations with 95% confidence
(no. of deaths/
2.4:2.4 2.212.2 2.0 : 2.0 1.8:1.8 1.6: 1.6
3,o:o 2,5:0 2.0:0 1,5:0
and LD5,s tests
LD,, for Mortality
Single-dose 7110 9110 S/IO O/IO l/l0 O/IO
6.0:0 5.0:0 4.0:0 3.0:0 2.0:0 1.0:0
B. YAGEN
l/8 O/S 218 018 O/8 2/s
0:5.0 (0:4.03-6.20)
test 2.90:0 (2,3&3.57:0)
2.38 :0.88 (2.14-2.64:0.66-1.17)
1.55: 1.55 (l,38-1,74:1,38-1.74)
0.88 : 2.38 (0.64-1.21 :2.13-2.67)
W3 2110 l/IO 2/10
0:4,15 (0:3,29-5.23)
birds typically lost Ion/, of their body weight and had clear or white opaque liquid faecal droppings. Decreased spontaneous activity was followed by sternal recumbency and death. The single oral dose LD50 determinations are presented in Table 1. From these data plotted on an isobologram (Fig. 1). it was concluded that T-2 toxin and diacetoxyscirpenol have additive lethal toxicity when dosed in combination. The dashed line in Fig. 1 represents the line of additive action and the three combination LDso values lie on either side of this line, a feature of additive action.
T-2 toxin and diacetoxyscirpenol
4
1
2 T-2
Toxin,
3
4
mg/ kg
Fig. I. Isobologram of the joint potency of T-2 toxin and diacetoxyscirpenol as defined by the single oral dose LDSo. The dashed line represents assumed additive toxicity.
Muh’ple-dose
toxicity
Chicks that died of multiple doseshad clinical signs similar to those described in the single-dose study. Most of the birds that died succumbed by the seventh day. The surviving chicks had decreases of body weight gain and packed cell volume. The LD,, values for the multiple dose study are presented in Table 1. Application of the isobologram method of analysis (Fig. 2) to these data indicates additive lethal toxicity for T-2 toxin and diacetoxyscirpenol when administered in combinations as 14 consecutive daily oral doses. DISCUSSION
This study defined the lethal effects of two naturally occurring trichothecene mycotoxins, T-2 toxin and diacetoxyscirpenol, administered in combination. The results indicated additive toxicity and should be useful in the correlation of severity of clinical disease with data obtained from the analysis of toxic feedstuffs. There are several reasons, however, why the results should be interpreted within the limits of conditions of this study. Only male chicks were used and sex has been shown to be an important variable when evaluating the interactions of mycotoxins. Thacker (1976) concluded that ochratoxin A and citrinin had synergistic lethal toxicity in female guinea-pigs but the effects were additive in males. This may be important for trichothecene mycotoxins because of their natural occurrencein association with zearalenone (Ghosal et a/. 1978; Mirocha et al. 1979) a commonly encountered mycotoxin with oestrogenic biological activity.
toxicity
187
Secondly, the interaction of T-2 toxin and diacetoxyscirpenol was defined only by the lethal toxicity. The sublethal toxicity of two mycotoxins in combination can be more complex. Sterigmatocystin and aflatoxin Br administered in combination to guinea-pigs caused a synergistic increase in the serum albumin, but decreases of a,-globulin, B-globulin, and complement activity were additive effects (Richard, Thurston, Lillehoj, Cysewski & Booth, 1978). The contributions of these various factors could modify the severity of clinical disease. A third reason involves understanding the role of mycotoxins and their contributions to diseasein the presence of other important factors. Hamilton (1977) reviewed substantial data and concluded that mycotoxins can have synergisticdetrimental effects in combination with nutritional deficiencies. The same may also hold true for infectious diseases, an area of special importance for the trichothecene mycotoxins because of their detrimental effects on the immune system (Boonchuvit, Hamilton & Burmeister, 1975). Thus, the effects of one or more mycotoxins in feedstuffs must be evaluated with regard to possible concurrent nutritional or infectious diseases. T-2 toxin was more toxic than diacetoxyscirpenol in both the single- and multiple-dose tests conducted in this study. These results differ from previously reported data that showed diacetoxyscirpenol to be more toxic than T-2 toxin in chickens (Chi, Robison, Mirocha & Reddy, 1978). The two derivatives differ structurally only in that T-2 toxin possessesan isovaleroxy group at carbon-8 and diacetoxyscirpenol has an Hz group in that position (Mirocha, 1979). While the differences in toxicity are small, they indicate that
\ 4 -‘\
g 3\ P \ F 5 6P
\
\
\
\
\
\
\
\
\
.
\
\
\
\ \
2-
\
‘Z B
. ‘\
\
i!! B
\ \ \
l-
\. \ \ \ \ \ \I
0 0
2
1 T-2
Toxin,
3
mg/kg/dw
Fig. 2. Isobologram of the joint potency of T-2 toxin and diacetoxyscirpenolas defined by the ICday multiple oral dose LD,,. The dashed line representsassumedadditive toxicity.
F. J. HOERR,
188
W. W. CARLTON
the functional group at the C-S position may have a role in the biotransformation of’ihe trichothecenes in chickens, as was found in rabbits (Ohta, Matsumoto, Ishii & Ueno, 1978). The metabolic role of the specific groups positioned at C-8 could, however, be different for the two species. In rabbits, the C-8 moieties of T-2 toxin and diacetoxyscirpenol did not affect de-acetylation of the C-4 position by hepatic carboxylesterase, which is the major mechanism of biotransformation, This type of comparative study has not yet been published for chickens. Acknowledgements-This work was supported by a Lilly Fellowship in Veterinary Pathology sponsored by Eli Lilly Co. and was published as Journal Paper No. 8085 of the Purdue University Agricultural Experiment Station, West Lafayette, Indiana 47907.
and B.
Hamilton, P. B. (1977). Interrelationships of mycotoxins with nutrition. Fedn Proc. Fedn Am. Sots exp. Eiol. Med. 36, 1899. Hewlett. P. S. (1969). Measurement of the potencies of drug mixtures. Biometrics 25, 477. Litchfield. J. T., Jr. & Wilcoxon, F. (1949). A simplified method of evaluating dose-effect experiments. J. Pharmm. exp. Ther. %. 99. Mirocha, .C. J. (1979). Trichothecene toxins produced by Fusarium. In Conierence on Mycoroxins it1 Animal Feeds und Grains Related to Animul He&h. Edited by W. Shimoda. p. 288. Report No. FDA/BVM-79/139. Mirocha. C. J.. Schauerhamer, B., Christensen, C. M. & Kommedahl. T. (1979). Zearalenone, deoxynivalenol, and T-2 toxin associated with stalk-rot in corn. Appl. envir.
Microbial.
REFERENCES
38, 557.
Ohta. M. Matsumoto, H.. Ishii. K. & Ueno. Y. (1978). Metabolism of trichothecene mycotoxins. II. Substrate specificity of microsomal deacetylation of trichothecenes. J. Biochem.
Boonchuvit, B., Hamilton, P. B. & Burmeister. H. R. (1975). Interaction of T-2 toxin with Salmonella infections in chickens. Poulrrv Sci. 54. 1693. Chi, M. S., Robison, T. S...Mirocha, C. J. & Reddy, K. R. (1978). Acute toxicity of 12,13-epoxytrichothecenes in one-day-old broiler chicks. Appl. enuir. Microhiol. 35, 636. Ciegler, A. (1978). Trichothecenes: occurrence and toxicoses. J. Fd Prof. 41, 399. Ghosal, S., Biswas. S., Srivastava, R. S., Chakrabarti, D. K. & Basu Chaudhary, K. C. (1978). Toxic substances produced by Fusarium V: Occurrence of zearalenone. diacetoxyscirpenol, and T-2 toxin in moldy corn infected with Fusarium monilforme Sheld. J. phorm. Sci. 67. 1768.
YAGEN
Jap.
84, 697.
Richard, J. L.. Thurston, J. R., Lillehoj. E. B., Cysewski. S. J. & Booth. G. D. (1978). Complement activity. serum protein, and hepatic changes in guinea pigs given sterigmatocystin or allatoxin. alone or in combination. Am. J. uer. R&. 39, 163. Sansine. G. A.. Lillehoi. E. B.. Detrov. R. W. & Miller, M. A’. (1976). Synergistic toxic effects of citrinin, ochratoxin A and penicillic acid in mice. Toxicon 14, 213. Thacker, H. L. (1976). Citrinin and ochratoxin A mycotoxicosis in guinea-pigs. p. 207. PhD Dissertation, Purdue University, West Lafayette, IN. Ueno, Y., Ishii. K., Sawano, M., Ohtsubo, K., Matsuda, Y.. Tanaka, T.. Kurata, H. & Ichinoe. M. (1977). Toxicological approaches to the metabolites of Fusoriu. XI. Trichothecenes and zearalenone from Fustrrium species isolated from river sediments. Jcrp. J. e.\p. Med. 47, 177.
Fd Cosmrr. Toricol. Vol. 19, pp. I85 10 188. 1981 Printed in Great Britain. All rights reserved
THE TOXICITY OF T-2 TOXIN AND DIACETOXYSCIRPENOL IN COMBINATION FOR BROILER CHICKENS F.J. HOERR and W.W. CARLTON Department
of Veterinary
Microbiology, Pathology and Purdue University, Lafayette,
Public Health, IN
47907,
School USA
of Veterinary
Medicine,
and B. YAGEN Department
01 Natural
Products,
School Jerusalem,
(Received
of Pharmacy, Israel
2SAugust
The Hebrew
University,
1980)
Abstract-T-2 toxin and diacetoxyscirpenol,two naturally occurring trichothecenemycotoxins,were dissolved in dimethylsulphoxide:saline (1:9, v/v) and administered separately and in combinations to 7-day-old male broiler chickens. The 72-hr single oral dose LD,, for T-2 toxin was 4.0mg/kg body .weight and for diacetoxyscirpenol. it was 5.0 mg/kg. T-2 toxin administered as 14 daily oral doses had an LDse of 2,90mg/kg, and diacetoxyscirpenol had an LD,, of 4.15 mg/kg. Combinations of the toxins caused additive lethal effects in both the single- and multiple-dose tests.
INTRODUCTION The 12,13-epoxy-trichothecenes are a group of at least 40 structurally related toxic fungal metabolites, most of which are produced by Fusarium species (Mirocha, 1979). Four of these mycotoxins, T-2 toxin, diacetoxyscirpenol, deoxynivalenol and nivalenol, have been identified as toxic components of naturally contaminated feedstuffs. Of these four toxins, two have been found in a single feed sample on at least two occasions (Ghosal, Biswas, Srivastava, Chakrabarti & Basu Chaudhary, 1978; Mirocha, Schauerhamer, Christensen & Kommedahl, 1979),and several species of Fusarium can produce combinations of these and other trichothecenes when cultured under laboratory conditions (Ciegler, 1978; Ueno, Ishii, Sawano, Ohtsubo. Matsuda, Tanaka, Kurata & Ichinoe, 1977). Synergistic lethal toxicity of three metabelites of Penicillium uiridicatum (ochratoxin A, citrinin, and penicillic acid) was demonstrated in female mice (&sing, Lillehoj, Detroy 8~ Miller, 1976). This paper describes the toxicity of T-2 toxin and diacetoxyscirpenol, given separately and in combinations as single and multiple doses, in male broiler chickens. EXPERIMENTAL
Chickens, Male broiler chicks of the Hubbard x Hubbard strain were obtained when one day old from a commercial hatchery (Fairview Farms, Remington, IN). The parent flock was seronegative to Myc@ plasma gallisepticum and M. synouiae, and the chicks received Marek’s disease vaccine prior to shipment. They were reared in wire-floored brooding cages measuring 60 x 90 x 20cm, with a brooding temperature of 3540°C a room temperature of 22°C continuous illumination, and forced air ventilation. A maximum of 50 birds were housed in each cage until 185
they were 14 days old when the maximum density was reduced to 32 birds per cage. Non-medicated commercial starter mash for broiler chicks and fresh tap water were available ud lib. The chicks were acclimatized and observed daily but not handled until they were 7 days old, when the body weight was about 100g. Sequentially numbered aluminium wingbands were randomized, affixed to the wing web, and the birds were placed in dose groups according to the wingband number sequence. Toxins and solvent. T-2 toxin was prepared by B. Yagen and diacetoxyscirpenol was obtained from Makor Chemical, Ltd. (Jerusalem, Israel). Each toxin was dissolved in one part reagent-grade dimethylsulphoxide and then diluted to a final volume with nine parts of physiological saline to make stock solutions of appropriate concentrations for administration to the high-dose groups. These stock toxin solutions were diluted to the desired concentrations with a stock solvent solution of dimethylsulphoxide:saline (1:9 v/v). The solvent exposure was 10 ml/kg body weight regardlessof the toxin dose. For the multiple dose study, the toxic solutions were prepared daily and administered within 3 hr of preparation. Toxin administration. Single oral dose 72-hr LDs, determinations were made for each toxin given separately and in dosage combinations presented in Table 1. The toxins were administered into the crop using a l-ml plastic tuberculin syringe and a curved 75cm dosing needle. Feed was withheld 4 hr prior to dosing The LD,e was calculated by the method of Litchfield & Wilcoxon (1949) and the joint potency was defined by the isobologram method of Hewlett (1969) for two drugs of separate activity. Ten birds were in each dose group including a solvent-only group. Multiple oral dose LD,, determinations were based on 14 consecutive daily doses administered to dose groups described in Table 1. The method of
186
F. J. HOERR. Table
W. W. CARLTON
and
1. T-2 toxin
and diacetoxyscirpenol single and Zl-day multiple
73hr
combinations oral toxicity
T-2 toxin: diacetoxyscirpenol dosages
(mg/kg body weight)
4.5 : I .75 4.0: 1.5 3.5:1.25 3.0: 1.0
no. in group)
4.0:0 (2.74-5.84:O)
3.25: 1.14 (3.01-3.51 : 1.06-1.22)
S/IO 4110 4/10 l/l0 l/l0
2.2612.26 (I ,74-2.94: 1.74-2.94)
IO/IO IO/IO S/IO S/IO
0:60 0:5.0 0:4.0 0:3.0 0:2.0 0: 1.0
1.03 : 3.05 (0.82-1~28:244-3.81)
4110 5110 5110 o/to O/IO O/IO Multiple-dose 5110 4110 O/IO O/IO
2.50: I.0 2.25 ~0.75 2GO:0~50 1.75:@25
518 3/8
1.75: 1.50: 1.25: 1.00:
1.75 1.50 1.25 I.00
5/8 318
1.0:2.50 0.75 12.25 0~50:2GG 0.25: 1.75
518
0:3.5 0:3,0 0:2.5
toxin administration was similar to that in the singledose study except that feed was withheld 2 to 3 hr prior to dose administration. For individual toxin administration and solvent-only controls, there were ten birds per dose group. Eight birds were in each group that received the combined toxins. RESULTS toxicity
The toxins administered alone and in combinations usually caused death within 24 hr of dosing. AfTected
intervals
test
IO/10 S/IO S/IO 4/10
1.75:4.5 1.5:4.0 1.25:3.5 1.0:3.0
Single-dose
for
toxin combinations with 95% confidence
(no. of deaths/
2.4:2.4 2.212.2 2.0 : 2.0 1.8:1.8 1.6: 1.6
3,o:o 2,5:0 2.0:0 1,5:0
and LD5,s tests
LD,, for Mortality
Single-dose 7110 9110 S/IO O/IO l/l0 O/IO
6.0:0 5.0:0 4.0:0 3.0:0 2.0:0 1.0:0
B. YAGEN
l/8 O/S 218 018 O/8 2/s
0:5.0 (0:4.03-6.20)
test 2.90:0 (2,3&3.57:0)
2.38 :0.88 (2.14-2.64:0.66-1.17)
1.55: 1.55 (l,38-1,74:1,38-1.74)
0.88 : 2.38 (0.64-1.21 :2.13-2.67)
W3 2110 l/IO 2/10
0:4,15 (0:3,29-5.23)
birds typically lost Ion/, of their body weight and had clear or white opaque liquid faecal droppings. Decreased spontaneous activity was followed by sternal recumbency and death. The single oral dose LD50 determinations are presented in Table 1. From these data plotted on an isobologram (Fig. 1). it was concluded that T-2 toxin and diacetoxyscirpenol have additive lethal toxicity when dosed in combination. The dashed line in Fig. 1 represents the line of additive action and the three combination LDso values lie on either side of this line, a feature of additive action.
T-2 toxin and diacetoxyscirpenol
4
1
2 T-2
Toxin,
3
4
mg/ kg
Fig. I. Isobologram of the joint potency of T-2 toxin and diacetoxyscirpenol as defined by the single oral dose LDSo. The dashed line represents assumed additive toxicity.
Muh’ple-dose
toxicity
Chicks that died of multiple doseshad clinical signs similar to those described in the single-dose study. Most of the birds that died succumbed by the seventh day. The surviving chicks had decreases of body weight gain and packed cell volume. The LD,, values for the multiple dose study are presented in Table 1. Application of the isobologram method of analysis (Fig. 2) to these data indicates additive lethal toxicity for T-2 toxin and diacetoxyscirpenol when administered in combinations as 14 consecutive daily oral doses. DISCUSSION
This study defined the lethal effects of two naturally occurring trichothecene mycotoxins, T-2 toxin and diacetoxyscirpenol, administered in combination. The results indicated additive toxicity and should be useful in the correlation of severity of clinical disease with data obtained from the analysis of toxic feedstuffs. There are several reasons, however, why the results should be interpreted within the limits of conditions of this study. Only male chicks were used and sex has been shown to be an important variable when evaluating the interactions of mycotoxins. Thacker (1976) concluded that ochratoxin A and citrinin had synergistic lethal toxicity in female guinea-pigs but the effects were additive in males. This may be important for trichothecene mycotoxins because of their natural occurrencein association with zearalenone (Ghosal et a/. 1978; Mirocha et al. 1979) a commonly encountered mycotoxin with oestrogenic biological activity.
toxicity
187
Secondly, the interaction of T-2 toxin and diacetoxyscirpenol was defined only by the lethal toxicity. The sublethal toxicity of two mycotoxins in combination can be more complex. Sterigmatocystin and aflatoxin Br administered in combination to guinea-pigs caused a synergistic increase in the serum albumin, but decreases of a,-globulin, B-globulin, and complement activity were additive effects (Richard, Thurston, Lillehoj, Cysewski & Booth, 1978). The contributions of these various factors could modify the severity of clinical disease. A third reason involves understanding the role of mycotoxins and their contributions to diseasein the presence of other important factors. Hamilton (1977) reviewed substantial data and concluded that mycotoxins can have synergisticdetrimental effects in combination with nutritional deficiencies. The same may also hold true for infectious diseases, an area of special importance for the trichothecene mycotoxins because of their detrimental effects on the immune system (Boonchuvit, Hamilton & Burmeister, 1975). Thus, the effects of one or more mycotoxins in feedstuffs must be evaluated with regard to possible concurrent nutritional or infectious diseases. T-2 toxin was more toxic than diacetoxyscirpenol in both the single- and multiple-dose tests conducted in this study. These results differ from previously reported data that showed diacetoxyscirpenol to be more toxic than T-2 toxin in chickens (Chi, Robison, Mirocha & Reddy, 1978). The two derivatives differ structurally only in that T-2 toxin possessesan isovaleroxy group at carbon-8 and diacetoxyscirpenol has an Hz group in that position (Mirocha, 1979). While the differences in toxicity are small, they indicate that
\ 4 -‘\
g 3\ P \ F 5 6P
\
\
\
\
\
\
\
\
\
.
\
\
\
\ \
2-
\
‘Z B
. ‘\
\
i!! B
\ \ \
l-
\. \ \ \ \ \ \I
0 0
2
1 T-2
Toxin,
3
mg/kg/dw
Fig. 2. Isobologram of the joint potency of T-2 toxin and diacetoxyscirpenolas defined by the ICday multiple oral dose LD,,. The dashed line representsassumedadditive toxicity.
F. J. HOERR,
188
W. W. CARLTON
the functional group at the C-S position may have a role in the biotransformation of’ihe trichothecenes in chickens, as was found in rabbits (Ohta, Matsumoto, Ishii & Ueno, 1978). The metabolic role of the specific groups positioned at C-8 could, however, be different for the two species. In rabbits, the C-8 moieties of T-2 toxin and diacetoxyscirpenol did not affect de-acetylation of the C-4 position by hepatic carboxylesterase, which is the major mechanism of biotransformation, This type of comparative study has not yet been published for chickens. Acknowledgements-This work was supported by a Lilly Fellowship in Veterinary Pathology sponsored by Eli Lilly Co. and was published as Journal Paper No. 8085 of the Purdue University Agricultural Experiment Station, West Lafayette, Indiana 47907.
and B.
Hamilton, P. B. (1977). Interrelationships of mycotoxins with nutrition. Fedn Proc. Fedn Am. Sots exp. Eiol. Med. 36, 1899. Hewlett. P. S. (1969). Measurement of the potencies of drug mixtures. Biometrics 25, 477. Litchfield. J. T., Jr. & Wilcoxon, F. (1949). A simplified method of evaluating dose-effect experiments. J. Pharmm. exp. Ther. %. 99. Mirocha, .C. J. (1979). Trichothecene toxins produced by Fusarium. In Conierence on Mycoroxins it1 Animal Feeds und Grains Related to Animul He&h. Edited by W. Shimoda. p. 288. Report No. FDA/BVM-79/139. Mirocha. C. J.. Schauerhamer, B., Christensen, C. M. & Kommedahl. T. (1979). Zearalenone, deoxynivalenol, and T-2 toxin associated with stalk-rot in corn. Appl. envir.
Microbial.
REFERENCES
38, 557.
Ohta. M. Matsumoto, H.. Ishii. K. & Ueno. Y. (1978). Metabolism of trichothecene mycotoxins. II. Substrate specificity of microsomal deacetylation of trichothecenes. J. Biochem.
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