We investigated whether direct injection of hemostatic agents into injury sites under the guidance of contrast-enhanced ultrasound (CEUS) can decrease blood loss from liver trauma. Twelve adult healthy dogs underwent laparotomy, and a hepatic wound (6.0 x 4.5 cm) was created in each dog. All animals were divided randomly into two groups: the treatment group and the control group. In the treatment group, hemocoagulase Atrox (Solco Basle Ltd., Birsfelden, Switzerland) and absorbable cyanoacrylate were injected directly into the liver injury sites, and 0.9% normal saline was injected into the injury sites in the control group. Resuscitation volume in the treatment group was less than that in the control group (55 +/- 12 mL vs. 105 +/- 23 mL, p < 0.001). The control group presented bleeding from liver injury sites. Times of bleeding at the injury site in the treatment groups were 6 +/- 1 s. Both groups presented an-echo and/or hypo-echo contrast regions, with contrast material pooling in the injury sites before the therapy. After the injection, the contrast material pooling disappeared in the treatment group observed by CEUS. Gross examination of liver tissue revealed that the injury site that had been treated was covered by clot and glue membrane. Histopathologic examination showed that there was hemostitic glue embolizing the microvessels and inflammatory cell infiltration among hepatocytes. Direct injection of hemocoagulase Atrox and cyanoacrylate into the liver injury sites guided by CEUS can effectively reduce blood loss from liver trauma within a short time. This experimental study may suggest a possible treatment for liver trauma hemorrhage and provide an experimental evidence for treating blunt abdominal parenchyma organs trauma under the guidance of CEUS.