The use of Chenopodium ambrosioides L. has shown to be promising in the management of gastrointestinal nematodes. The objective of this study was to quantitate the yield and characterize the chemical composition of the essential oil of C. ambrosioides, as well as to evaluate the in vitro effect of the ethanolic extract and the essential oil in L3 of Ancylostoma spp. and the in vivo effect(s) of the essential oil in dogs. The effects of the ethanol extract and essential oil on Ancylostoma spp. were evaluated in vitro by exposing larvae to the extract at concentrations ranging from 0.005 g mL -1 to 0.2 g mL -1 and to essential oil at concentrations of 50, 100, and 150 μL mL -1 . For the in vivo test, 26 healthy dogs, naturally infected by Ancylostoma spp., were divided into three groups: F1 -cookies were administered without active principle; F2 -herbal cookies containing 37.5 μL g -1 essential oil of C. ambrosioides L.; F3 -cookies plus a commercial formulation containing febantel, pyrantel, praziquantel, and ivermectin. Complete blood counts and serum biochemistry for AST, ALT, AF, urea, creatinine, total protein, and albumin were performed. The yield of the essential oil was 0.3% m v -1 , and its major components included α-terpinene (1.24%), p-cymene (4.83%), and (Z)-ascaridole (87%) and E (5.04%) isomers. The concentrations of C. ambrosioides L. ethanol extract used were ineffective against Ancylostoma spp. larvae. The essential oil at a concentration of 150 μL mL -1 was effective against L3 larvae. In the in vivo study in dogs, the herbal cookies containing C. ambrosioides L essential oil reduced the number of eggs per gram of feces. g mL -1 e 0,2 g mL -1 , do óleo essencial nas concentrações de 50 µL mL -1 , 100 µL mL -1 e 150 µL mL -1 . Para o teste in vivo utilizaram-se 26 cães, hígidos, infectados naturalmente por Ancylostoma spp., divididos em três grupos: F1 -foram administrados biscoitos sem princípio ativo; F2 -biscoitos fitoterápicos com óleo essencial de C. ambrosioides L na concentração de 37,5 µL g -1 ; F3 biscoitos acrescido de formulação comercial a base de febantel, pirantel, praziquantel e ivermectina. Foi realizado hemograma completo e bioquímica sérica para AST, ALT, FA, ureia, creatinina, proteína total e albumina. Verificouse o rendimento de 0,3% m v -1 do óleo essencial e na sua composição química a presença de α-terpineno (1,24 %), o p-cimeno (4,83 %) e ascaridol, nos seus isômeros Z (87%) e E (5,04 %), como componente majoritário. O extrato etanólico de C. ambrosioides L., nas concentrações utilizadas, apresentou insuficiente atividade para o efeito larvicida. O óleo essencial na concentração de 150 µL mL -1 foi eficaz contra L 3 . A formulação de biscoito fitoterápico com óleo essencial C. ambrosioides L reduziu o número de ovos por grama de fezes dos cães. Palavras-chave: Erva-de-santa-maria. Helmintos. Óleo essencial. Planta medicinal.