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in Journal of the American Veterinary Medical Association



To examine the efficacies of combinations of 7 sulfonamides and 5 dihydrofolate reductase/thymidylate synthase (DHFR/TS) inhibitors against tachyzoites of Neospora caninum in cultured cells. Mutant tachyzoites that were resistant to pyrimethamine were produced and examined for resistance to other DHFR/TS inhibitors.

Design and Procedures

After 5 days of treatment, a cell culture flask lesion-based assay was used to determine efficacies of combinations of sulfonamides and DHFR/TS inhibitors against N caninum tachyzoites and to evaluate the sensitivity of pyrimethamine-resistant mutants of N caninum to test agents. Cultured cells that were infected with the appropriate strains of N caninum and treated or not treated (controls) with test agents were examined. Mutations were induced by chemical mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidme or by selection for growth in permissive concentration of pyrimethamine.


Synergism was detected for combinations of pyrimethamine, ormetoprim, trimethoprim, or diaveridine with the sulfonamides. Methotrexate did not have improved efficacy when combined with sulfonamides. Two mutants were produced that were resistant to pyrimethamine. Both mutants were resistant to other DHFR/TS inhibitors. Both mutants remained resistant to pyrimethamine in the absence of continuous exposure to the agent, indicating that the induced resistance was stable. Synergism was detected for combinations of DHFR/TS inhibitors and sulfonamides against these pyrimethamine-resistant mutants.


Combinations of suboptimal concentrations of sulfonamides with suboptimal concentrations of DHFR/TS inhibitors results in improved efficacy of the agents in a cell culture assay. Stable resistance to pyrimethamine can be induced in N caninum tachyzoites by use of chemical mutagenesis or by selection.

Clinical Relevance

In vitro evidence indicated that combination treatment, using sulfonamides and DHFR/TS inhibitors, may be effective in treating neosporosis. (Am J Vet Res 1996;57:68-72)

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in American Journal of Veterinary Research



To identify the lowest single dose of lufenuron injected SC that results in a 90% disruption of the flea (Ctenocephalides felis) life cycle for 6 months in cats.


40 domestic shorthair cats (20 males, 20 females) between 5 and 7 months old.


Cats were randomly assigned to 1 of 5 eight-cat groups and experimentally infested with C felis on days −8, −7, −6, and −4. On day 0, cats in the 4 treatment groups were treated with an injectable formulation of lufenuron at doses of 2.5, 5, 10, or 20 mg/kg of body weight, respectively. Control cats received the injectable formulation without lufenuron. Experimental infestations were repeated and flea eggs collected at various intervals for 196 days after treatment. Eggs were placed in media and incubated in an insectary for 28 days to determine effects of injectable lufenuron on egg and larval development. Number of adults that emerged from eggs were compared among groups.


Lufenuron injected once at a dose of 10 or 20 mg/kg, but not at 2.5 or 5 mg/kg, resulted in a 90% decrease in number of adult fleas emerging from eggs for 196 days after treatment.

Conclusions and Clinical Relevance

Results indicate that control of flea egg and larval development for at least 6 months can be achieved in cats with a single SC injection of lufenuron (10 mg/kg). The injectable formulation may provide veterinarians and cat owners an alternative to the tablet formulation of lufenuron. (Am J Vet Res 1999;60:1513–1515)

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in American Journal of Veterinary Research