Psoroptes ovis var. cuniculi is a common ectoparasite of the wild and domestic rabbits worldwide, which causes significant economic losses in commercial rabbit breeding. In China, the diagnosis of rabbits infested with P. ovis var. cuniculi currently relies on detection of clinical signs and Psoroptes mites in skin scrapings by microscopy examination. However, this method is not very efficient in detection of the low mite loads and/or sub-clinical infections. In the present study, we cloned and expressed an arginine kinase homolog gene from P. ovis var. cuniculi (Poc-AK) and used its recombinant protein rPoc-AK to develop an indirect enzyme-linked immunosorbent assay (iELISA) method for diagnosis of P. ovis var. cuniculi infestation in rabbits. The results showed that the rPoc-AK protein was ∼61 kDa and had no signal peptide. The rPoc-AK-based iELISA achieved a 94.4% sensitivity and a 88.2% specificity, and was able to detect P. ovis var. cuniculi infection as early as the 1st week post-infection, prior to the appearance of clinical signs. Further field study showed 24.94% (66.33/266) clinically normal rabbits were seropositive with the highest and lowest seropositive rates for California (35.71%) and Belgian (15.14%), respectively. These results suggested that the rPoc-AK has potential as a diagnostic antigen for early P. ovis var. cuniculi infestation in rabbits.

BackgroundSarcoptic mange caused by the mite Sarcoptes scabiei is a worldwide disease affecting both humans and animals. Here we report the molecular characterization and evaluation of a recombinant S. scabiei tropomyosin (SsTm) protein in a vaccination trial in rabbits.MethodsThe full-length cDNA was cloned in a bacterial pET vector, and the recombinant protein was expressed in BL21 (DE3) cells and purified. Using specific rabbit antiserum, tropomyosin was localized immunohistochemically in mite tissue sections. Vaccination trials with the recombiant SsTm was carried out in New Zealand rabbits.ResultsThe full-length open reading frame (ORF) of the 852 bp cloned gene from S. scabiei encodes a 32.9 kDa protein. The amino acid sequence showed 98.94%, 97.89% and 98.59% homology to Dermatophagoides farina and Dermatophagoides pteronyssinus group 10 allergens and Psoroptes ovis tropomyosin, respectively. Tropomyosin was localized immunohistochemically in mite tissue sections mainly in the mouthparts, legs and integument of the epidermis. The predicted cross-reactivity of SsTm indicated that it is an allergenic protein. While vaccination with the recombiant SsTm resulted in high levels of specific IgG (P < 0.01), a low IgE antibody response and no significant protection against S. scabiei challenge were observed. After challenge, specific IgG levels remained significantly higher than the control (P < 0.01), while changes of total IgE levels were not significant (P > 0.05). However, the lesion areas in the vaccination group decreased at the end of the experiment compared with controls.ConclusionsAlthough vaccination with recombinant SsTm did not efficiently control sarcoptic mange in rabbits, the immunogenic properties of tropomyosin suggest it may be developed as a vaccine with alternative adjuvants or delivery methods.

BackgroundScabies is caused by Sarcoptes scabiei burrowing into the stratum corneum of the host’s skin and is detrimental to the health of humans and animals. Vaccines are an attractive alternative to replace the acaricides currently used in their control.MethodsIn the present study, the S. scabiei chitinase-like protein 5 (SsCLP5) was characterized and recombinant SsCLP5 (rSsCLP5) was evaluated as a candidate vaccine protein for anti-mite protection in rabbits. The expression, characterization and immunolocalization of SsCLP5 were examined. Vaccination experiments were performed on three test groups (n = 12 per group) immunized with purified rSsCLP5. Control groups (n = 12 per group) were immunized with PBS, QuilA saponin or empty vector protein. After challenge, the inflammatory reaction and skin lesions were graded and rSsCLP5 indirect ELISA was used to detect antibody IgG levels in serum samples at the time of vaccination and post-challenge.ResultsThe results showed that rSsCLP5 had high immunoreactivity and immunogenicity. In S. scabiei, SsCLP5 had a wide distribution in the chewing mouthpart, legs and exoskeleton, especially the outer layer of the exoskeleton. Vaccination with rSsCLP5 resulted in 74.3% (26/35) of rabbits showing no detectable lesions after challenge with S. scabiei.ConclusionsOur data demonstrate that rSsCLP5 is a promising candidate for a recombinant protein-based vaccine against S. scabiei. This study also provides a method for studying scabies vaccine using rabbit as an animal model and a basis for screening more effective candidate proteins.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-3184-y) contains supplementary material, which is available to authorized users.

The mite Psoroptes cuniculi is globally widespread and has a serious impact on commercial rabbit breeding. In China, diagnosis of P. cuniculi is currently based on conventional clinical methods that entail numerous disadvantages, including their failure to diagnose subclinical infections. Hence, alternative measures are required, and dot-ELISA is one of the most promising strategies. We cloned and expressed the recombinant P. cuniculi troponin C gene for use as a basis for novel dot-ELISA assay to detect P. cuniculi infections in rabbits. This amplified sequence encoded a 153 amino acid protein of 17·6 kDa and theoretical pI 4·18 without signal peptide. The recombinant troponin C of P. cuniculi is an outer membrane protein and may also be a new P. cuniculi allergen. Results of dot-ELISA test showed that this novel assay had more than 90% sensitivity but low specificity in distinguishing infections with P. cuniculi or Sarcoptic scabiei, despite very high agreement between observers (97-99%; κ values ranged from 0·95 to 0·98 for inter- and intra-observer variability test). This study showed that this novel method, at present, lacks diagnostic utility. Therefore, although simple serological assays such as dot-ELISA show great promise as diagnostic tools, we suggest that troponin C is not a suitable diagnostic antigen candidate.

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