Clinical efficacy of 9-oxo-10, 11-dehydroageraphorone extracted from Eupatorium adenophorum against Psoroptes cuniculiin rabbits
- Yang Hu†1,
- Fei Liao†1, 2,
- Yanchun Hu1Email author,
- Biao Luo1,
- Yajun He1,
- Quan Mo1,
- Zhicai Zuo1,
- Zhihua Ren1,
- Junliang Deng1 and
- Yahui Wei3
© Hu et al.; licensee BioMed Central Ltd. 2014
Received: 8 August 2014
Accepted: 13 December 2014
Published: 20 December 2014
Animal acariasis is one of the important veterinary skin diseases. Chemical drugs have been widely used to treat and control this kind of disease. But many chemicals control could increase resistance in target species, toxicity and environmental hazards. We found that the 9-oxo-10, 11-dehydroageraphorone (euptox A) extracted from E. adenophorum has strong toxicity against P. cuniculi in vitro, but the in vivo acaricidal actions of euptox A have yet to be investigated.
A 14-day experiment was performed using rabbits that were naturally infested with P. cuniculi on a farm. Rabbits were randomly divided into five groups; animals in groups A, B and C were treated in each ear topically with 4.0 ml of 2.0 and 1.0 g/L (w/v) euptox A, respectively. Animals in groups D and E were treated with ivermectin (by injection; positive controls) and glycerol with water only (by embrocation; negative controls), respectively. Each rabbit was treated twice with separate treatments on days 0 and 7. Rabbits were observed daily and detailed examinations were performed on days 0, 7 and 14, to inspect the presence or absence of mites and scabs/crusts. Seven days after the initial treatment, the mean clinical scores (presence of scabs/crusts) decreased from 3.48, 3.37, 3.43 and 3.45 to 0.37, 0.42, 0.78 and 0.38 in the ears of animals in groups A, B , C and D, respectively, which were similar to the observations recorded in the positive control rabbits. However, the clinical score for negative control rabbits did not increase significantly (P > 0.05) during the experiment, and this changed from 3.32 to 3.37 in the ears, and there were no significant differences in clinical efficacy between left and right ears. After two treatments (0 and 7 d), the rabbits in groups A, B, C and D had recovered completely 14 days after the last treatment and no recurrences of infection were observed.
These results indicate that euptox A was potent compounds for the effective control of animal P. cuniculi in vivo.
Animal acariasis, an important veterinary diseases, may reduce the productivity and the quality of animal products, even lead to death . At present, chemical drugs are widely used to treat and control the psoroptes and sarcoptic mange in veterinary clinic, and obtained the relative good treatment effectiveness, including ivermectin, and abamectin, etc. But the chemical control could increase resistance in target species, toxicity and environmental hazards .
A large number of reports have indicated that the secondary metabolites synthesized and accumulated in Eupatorium adenophorum (E. adenophorum) have wide biological activities. For example, there are reports of chronic respiratory disease and exercise intolerance in horses in Australia due to ingestion of E. adenophorum . The ethanol extract from leaves of E. adenophorum were anti-Inflammatory potential , acaricidal activity ,, antioxidant activity , and other extract form E. adenophorum torium had toxic activity against Oncomelania hupensis, the intermediate host snail of Schistosoma japonicum , Tinea  and Aphis gossypii . The acetone extract of E. adenophorum had strong toxicity against Cabbage aphids and Brevicoryne brassicae .
9-oxo-10, 11-dehydroageraphorone (euptox A), is the main toxin extracted from E. adenophorum ,. Euptox A takes a large proportion of E. adenophorum toxins ,, can cause not only the allergic bronchial pneumonia of horses which is characterized by pulmonary interstitial fibrosis, emphysema, alveolar epithelisation and reduced tolerance to exercise ,, but the contact dermatitis of other domestics animals like cattle and goats . Furthermore, according to some studies, for mice, lesions occur in the liver. The hepatic injury in these animals is characterised by multiple areas of focal necrosis of the parenchyma associated with degeneration and loss of the epithelium lining the small bile ducts ,. Euptox A belongs to a cadenine sesquiterpene. A large number of reports indicated that the cadenine sesquiterpene has wide biological activities such as antitumor activity , antigerminative activity , neurotrophic activity , larvicidal activity , antiprotozoal activity , and so on. Euptox A was found highly active against the fast growing A549, Hela and Hep-2, and its activity was concentration-dependent . In a previous study, euptox A exhibited strong toxicity against S. scabiei and P. cuniculi in vitro , but the in vivo acaricidal actions of euptox A have yet to be investigated.
Thus, the aim of this present study was to assess the clinical acaricidal efficacy of euptox A against P. cuniculi in rabbits in vivo.
Extraction and purification of euptox A
E. adenophorum was collected from Xichang City (102°30222′E, 27°873213′S) of Sichuan Province, Southwest China in July, 2012. Euptox A was extraction from E. adenophorum by the Key laboratory of Animal Disease and Human Health of Sichuan Agricultural University, China -.
Thirty rabbits that were naturally infected with P.cuniculi were obtained from a farm affected by an outbreak. All the animals were of similar age, weight and clinical score (in terms of the presence/absence of scabs and crusts). In all 30 rabbits, there were no significant difference between the level of infection in the ears, and had not been treated with acaricides and no other infectious diseases were known to be present. Sampling procedures adhered to institutional ethical and animal care guidelines and all methods were conducted in accordance with the Guide for the Care and Use of Laboratory Animals adopted and promulgated by the United National Institutes of health. All experimental protocols were approved by review committee for the use of human or animal subjects of College of veterinary medicine, Sichuan Agricultural University.
Acaricidal activity in vivo
The euptox A was diluted from the concentration of 4 mg/ml to 1 mg/ml (4 mg/ml, 2 mg/ml and 1 mg/ml) in 10% glycerin.
Parameters used to evaluate the clinical score of infection and degree of recovery
Infection and degree of recovery
Absence of scabs and/or mites
Irritation in ear canal but no mites observed
Small number of scabs in the ear canal, mites present
External ear canal filled with scabs, mites present
Scabs in ear canal and proximal 1/4 of pinna, mites present
1/2 pinna filled with scabs, mites present
3/4 of the pinna filled with scabs, mites present
All internal surface of the pinna full of scabs, mites present
Statistical analysis was performed using SPSS software (SPSS, version 20.0)  to assess the presence of statistically significant differences in infection and recovery scores, and therapeutic effects of different concentrations of euptox A and different treatment times. Significance values were corrected for with Duncan’s multiple comparisons test .
The clinical score of infection and degree of the recovery from infection of the external ear margins in rabbits (mean ± standard error)
3.48 ± 0.27A(A)
3.37 ± 0.21A(A)
3.43 ± 0.22A(A)
3.45 ± 0.23A(A)
3.23 ± 0.21A(A)
0.37 ± 0.19B(B)
0.42 ± 0.17B(B)
0.78 ± 0.15C(B)
0.38 ± 0.15B(B)
3.37 ± 0.23A(A)
0.00 ± 0.00B(C)
0.00 ± 0.00B(C)
0.00 ± 0.00B(C)
0.00 ± .0.00B(C)
3.45 ± 0.23A(A)
The effects of euptox A on rabbit itching mite negative transformation rate
Negative transformation rate %
24.33 ± 1.97
0.00 ± 0.00
0.00 ± 0.00
21.50 ± 2.17
0.00 ± 0.00
0.00 ± 0.00
18.61 ± 2.17
2.17 ± 1.72
0.00 ± 0.00
24.61 ± 2.07
0.00 ± 0.00
0.00 ± 0.00
19.83 ± 2.13
22.83 ± 2.48
24.00 ± 2.37
Good clinical efficacy was achieved with the euptox A extract from E. adenophorum, against the scab mites, P. cuniculi. The clinical acaricidal efficacy of euptox A was similar to that of injectable ivermectin, and this observation is consistent with the euptox A has strong toxicity against S. scabiei and P. cuniculi in vitro , whilst consistent with petroleum ether extract form E.adenophorum against P. cuniculi in vitro . The clinical acaricidal efficacy was showed to be time- and concentration-dependent, with the euptox A displaying similar effects to the alcohol extract from E. adenophorum against P. cuniculi. In the current study, the euptox A showed better clinical acaricidal efficacy (2ml/ml) than the alcohol extract from E. adenophorum (1 g/ml) , whilst the petroleum ether extract of neem oil comparatively weak bioactivity (LC50, 500.0 ll/ml) .
E. adenophorum has been reported to have hepatotoxic effects in rodents ,, which could limit its development as an acaricidal agent. But the acute toxicity test and skin hypersensitive test of euptox A had been finished, the euptox A were not irritant to the skin and toxicity of rabbits. Combining our previous results with this trial, we firmly believe that, euptox A shows a good clinical therapeutic effect on animal acariasis.
We believe that, after further in-depth study, euptox A, a potent herbal drug, will be more widely applied in treatments for humans and animals. This study provides a new way for utilization of E. adenophorum and the euptox A has the potential as acaricidal drugs in livestock, and future possible scope of the product in small animal vet medicine, e.g. against Octodectes ear mites and other common ectoparasites of dogs and cats. But in order to make the research systematic, the mechanism of action has yet to be determined.
Yang Hu and Fei Liao should be considered as first authors.
This research was supported by Science and Technology Support Program of Sichuan Province (Grant No. 2015SZ0201), Special Fund for Agroscientific Research in the Public Interest (Grant No. 201203062) and Chang-jiang Scholars and the Innovative Research Team in University (Grant No. IRT0848).
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