Abstract
There have been many reports concerning the toxicological effects of the alkylating agent, apholate (2, 2,4,4,6-hexahydro-2,2,4,4,6,6-hexakis (1-aziridinyl)-1,3,5,2,4,-6-triaza-triphosphorine) in insects. Toxicological studies have been conducted in cattle (1), sheep (2), rats (3), mice (4), and fowls (5,6), but work with fish has been limited (7).
The delayed lethal syndrome resulting from treatment of animals with alkylating agents was repoted by several workers, but the actual cause of death is not known (8). The present paper describes the hematolgical effects of oral administration of apholate to channel catfish Ictalurus punctatus), and the delay in lethality, even with massive doses.
Materids and Methods. Channel catfish fingerlings, weighing 5-20 g, were maintained at 22° in 5 or 10-gal aquariums 10-14 days before, and during, experimentation. Single oral doses of apholate (crystallized from ethyl acetate) were administered in no. 5 gelatin capsules.
In the study of hematological changes produced by apholate, total erythrocyte and leukocyte counts were made by the method of Hesser (9), and differential leukocyte counts were made on thin smears stained with Wright's stain, using the classification of Jakowska (10). The blood was obtained by cardiac puncture using heparin as anticoagulant. Preliminary (normal) counts were made on 90 animals, and 2 weeks were allowed for recovery from cardiac puncture before treatment. Individual fish were marked for identification by small notches in the caudal fin, so that cell counts on each animal could be compared. The animals were then divided into three groups at random; 30 controls received empty capsules, 30 animals received 200 mg of apholate/kg; and 30 animals received 4010 mg of apholate/ kg. At 3, 6, and 9 days after treatment, blood samples were obtained again from 10 animals from each group. The animals were sacrificed and sexed by microscopic examination of the gonads. Significance of changes in blood cell counts was tested by ranking.
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