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Background Psoroptes ovis var. cuniculi is a common ectoparasite of wild and domestic rabbits worldwide that causes economically devastating losses in commercial rabbit husbandry and significantly affects the overall health of rabbits. Serine proteinase inhibitor (serpin) is present in almost all organisms that are involved in host–pathogen interactions, inflammatory responses, and reproductive development, among others. However, very little research has been carried out on P. ovis var. cuniculi serpins. Methods Two serpin genes of P. ovis var. cuniculi (Pso c 27 and PsoSP2 cDNAs) were cloned and molecularly characterized. The transcriptional profiles and tissue localization of these two serpins in P. ovis var. cuniculi were investigated by quantitative real-time PCR and immunohistochemistry, respectively. The potential function of recombinant Pso c 27 and PsoSP2 (rPso c 27 and rPsoSP2) in the serodiagnosis of P. ovis var. cuniculi infestation in rabbits was evaluated using a newly devleoped indirect enzyme-linked immunosorbent assay. Results Both the 523-residue Pso c 27 and the 240-residue PsoSP2 proteins contained typical serpin domains and signatures. Both Pso c 27and PsoSP2 cDNAs were expressed throughout the life-cycle; specifically, the cDNAs showed significantly higher expression in female mites than in larva, nymph, and male mites (Pso c 27: F (3, 8)  = 1935.953, P  < 0.0001; PsoSP2: F (3, 8)  = 660.669, P  < 0.0001). The native Pso c 27 and PsoSP2 proteins localized in the ovary and mouthparts of adult female mites, respectively. Compared to rPsoSP2, rPso c 27 showed better diagnostic efficiency, with higher values of sensitivity, specificity, and area under the receiver operating characteristic curve (rPso c 27 vs rPsoSP2: 96.0 vs 90.0%; 90.91 vs 78.18%; 0.988 vs 0.964, respectively). Moreover, rPso c 27 showed seropositivity in 80% of the rabbits as early as the 2 weeks post-infestation, prior to visible clinical signs and microscopy-positive of skin scrapings. Conclusions These results suggest that these two serpins may play essential roles in reproductive development, serum-feeding, and pathogenicity of P. ovis var. cuniculi . Compared to PsoSP2, Pso c 27 appears to be a potential antigen for serodiagnosis of P. ovis var. cuniculi infestation in rabbits, especially at the early stage of infestation. Graphical Abstract

Background The ectoparasite Psoroptes ovis var. cuniculi causes substantial economic losses to the global rabbit industry. Currently, microscopy for identifying Psoroptes mite in skin scrapings, as the “diagnosis gold standard,” remains a challenge owing to its poor sensitivity in detecting low-level and/or early stage mite infestations. Additionally, Psoroptes infestations rapidly trigger cutaneous inflammation, thus the mites might produce some molecules to deal with the harmful effects of inflammation for their long-time survival on the host skin, but these molecules are still mostly unknown. Methods To seek a sensitive diagnostic method and illuminate the new antiinflammatory molecules, we characterized a novel cystatin of P. ovis var. cuniculi (PsoCys) using bioinformatics and molecular biology methods. Results The results showed that PsoCys comprised the classical features of the type II cystatin superfamily including an N-terminal glycine residue, a central QXVXG motif, and a C-terminal LW motif. In mixed stages of mites, the transcription level of PsoCys was significantly higher in “fed” mites than in “starved” mites ( P  < 0.001), and among the different life-cycle stages of “fed” mites, the expression of PsoCys was higher in adult males than in larva, nymph, and adult females ( P  < 0.001). The established indirect ELISA based on recombinant PsoCys (rPsoCys-iELISA) presented 95.4% sensitivity and 95.7% specificity. The area under the receiver operating characteristic curve (AUC) for this method was 0.991, indicating its excellent diagnostic performance. Moreover, rPsoCys-iELISA had advantages over microscopy for detecting low-level and/or early stage mite infestations (90% versus 40% in artificial infestation cases at 3 weeks post-infestation; 61.9% versus 22.6% in clinical cases). In addition, rPsoCys could inhibit the activity of papain and cathepsin B in vitro, and significantly suppressed mRNA levels of toll-like receptors (TLR 1, 2, 4, and 6) and downstream molecules (NF-κB, p38, MyD88, IL-10, and IFN-γ) in LPS-stimulated rabbit PBMCs, indicating its anti-inflammatory property. Conclusions Our findings indicated that PsoCys was a novel type II cystatin of Psoroptes mites, and it served as a potential serological diagnostic antigen for detecting low-level and/or early stage mite infestations, as well as a novel anti-inflammatory molecule of Psoroptes mites. Graphical abstract

Psoroptes ovis mites, the causative agent of sheep scab, can severely compromise sheep welfare and production. However, in subclinical infections, mite detection is difficult increasing the risk of spread. A recent serodiagnostic test, based on detecting host antibodies to the P ovis allergen, Pso o 2, has made the detection of subclinical infection possible. The use of this test was demonstrated in subclinical situations, through an opportunistic observational study on an extensive hill farm and a lowland flock with recently introduced, quarantined livestock. Twelve animals were tested from each group. Breeding ewes and lambs on the hill farm had seroprevalences of 16 per cent (12.5–17.8 per cent) and 8.3 per cent (4.8–10.1 per cent), respectively. Quarantined store lambs had a seroprevalence of 16.7 per cent (13.2–18.5 per cent); no evidence of P ovis was found in quarantined replacement ewes. By detecting subclinical infection, this serological test could be a powerful tool in sheep scab control, for quarantine procedures, accreditation programmes, and possibly regional or national eradication protocols.

Species delimitation of Psoroptes spp. and identity of the parasite in water buffaloes remain poorly defined. In this study, Psoroptes infestation on three water buffalo farms in Egypt was examined based on morphometric characteristics, especially the opisthosomal setae of adult male mites. Clinical investigations showed that 28% (196/700) of the sampled animals had mange infestation. Microscopic examinations of 80 skin scrapings indicated the occurrence of Psoroptes mites in 17 (21.3%) samples, Sarcoptes mites in 27 (33.7%) samples, and the concurrence of both in 36 (45.0%) samples. Morphologically, the Psoroptes parasite was identified as Psoroptes natalensis. DNA sequence analysis of the second internal transcribed spacer (ITS2) in 11 representative samples confirmed the diagnosis and suggested the presence of a distinct variety of Psoroptes natalensis in Egypt.

Ovine psoroptic mange (sheep scab) is an infection of substantial economic and animal welfare concern in the UK. Its prevalence has increased rapidly over the last 20 years and management is dependent on a small number of acaricidal compounds, many of which are also used to control a range of other endoparasites and ectoparasites. Here, the effects of the macrocyclic lactone (ML) moxidectin was considered using in vitro assays against mites from four farm populations where persistent treatment failure had been reported: two in West Wales, one from the England/Wales border and one in Herefordshire. The data demonstrate resistance in mites from all four farms. This is the first quantitative evidence of ML resistance in Psoroptes mites in the UK. Given the similarities in their mode of action it is highly likely that cross-resistance across the range of this class of compound will be found. The development of resistance to moxidectin is of considerable concern given the already high prevalence of scab infection in some regions; major difficulties in scab management should be anticipated if ML resistance becomes widely established in the UK.

BACKGROUND: Limited available sequence information has greatly impeded population genetics, phylogenetics and systematics studies in the subclass Acari (mites and ticks). Mitochondrial (mt) DNA is well known to provide genetic markers for investigations in these areas, but complete mt genomic data have been lacking for many Acari species. Herein, we present the complete mt genome of the scab mite Psoroptes cuniculi. METHODS: P. cuniculi was collected from a naturally infected New Zealand white rabbit from China and identified by morphological criteria. The complete mt genome of P. cuniculi was amplified by PCR and then sequenced. The relationships of this scab mite with selected members of the Acari were assessed by phylogenetic analysis of concatenated amino acid sequence datasets by Bayesian inference (BI), maximum likelihood (ML) and maximum parsimony (MP). RESULTS: This mt genome (14,247 bp) is circular and consists of 37 genes, including 13 genes for proteins, 22 genes for tRNA, 2 genes for rRNA. The gene arrangement in mt genome of P. cuniculi is the same as those of Dermatophagoides farinae (Pyroglyphidae) and Aleuroglyphus ovatus (Acaridae), but distinct from those of Steganacarus magnus (Steganacaridae) and Panonychus citri (Tetranychidae). Phylogenetic analyses using concatenated amino acid sequences of 12 protein-coding genes, with three different computational algorithms (BI, ML and MP), showed the division of subclass Acari into two superorders, supported the monophylies of the both superorders Parasitiformes and Acariformes; and the three orders Ixodida and Mesostigmata and Astigmata, but rejected the monophyly of the order Prostigmata. CONCLUSIONS: The mt genome of P. cuniculi represents the first mt genome of any member of the family Psoroptidae. Analysis of mt genome sequences in the present study has provided new insights into the phylogenetic relationships among several major lineages of Acari species.

Background Ovine psoroptic mange (sheep scab) is a highly pathogenic contagious infection caused by the mite Psoroptes ovis . Following 21 years in which scab was eradicated in the UK, it was inadvertently reintroduced in 1972 and, despite the implementation of a range of control methods, its prevalence increased steadily thereafter. Recent reports of resistance to macrocyclic lactone treatments may further exacerbate control problems. A better understanding of the factors that facilitate its transmission are required to allow improved management of this disease. Transmission of infection occurs within and between contiguous sheep farms via infected sheep-to-sheep or sheep–environment contact and through long-distance movements of infected sheep, such as through markets. Methods A stochastic metapopulation model was used to investigate the impact of different transmission routes on the spatial pattern of outbreaks. A range of model scenarios were considered following the initial infection of a cluster of highly connected contiguous farms. Results Scab spreads between clusters of neighbouring contiguous farms after introduction but when long-distance movements are excluded, infection then self-limits spatially at boundaries where farm connectivity is low. Inclusion of long-distance movements is required to generate the national patterns of disease spread observed. Conclusions Preventing the movement of scab infested sheep through sales and markets is essential for any national management programme. If effective movement control can be implemented, regional control in geographic areas where farm densities are high would allow more focussed cost-effective scab management. Graphical Abstract

Parasites and obesity are health problems worldwide. Rabbits are production animals yielding one of the healthiest meats, also taking advantage of skin, hair, limbs and excreta. It is among the most frequent pets in some countries and widely used as animal model in research. Psoroptes cuniculi is a mite of high transmission rate, affecting welfare and production and obesity causes multiple metabolic, endocrine and immunological disorders, being an emerging problem in domestic animals. Obesity and acarosis are prolonged stressors, modifying the activity of the hypothalamic-pituitary-adrenal axis that can induce metabolic and behavioral disorders. Alterations caused by comorbidities could be similar to or different from those induced by each morbidity separately. We analyzed the influence of obesity on the infection degree with P . cuniculi and on behavior and production. Rabbit does induced to obesity were infected and mated; behaviors in the open field test, obesity estimation indices and productive parameters at delivery and weaning were analyzed. The acarosis induced a decrease in feed intake and a decrease in body weight, a decrease in locomotor, exploratory and chinning behaviors in normal weight and obese does. The infection induced 23% mortality at birth, obesity 45% and comorbidity 74%, while in normal weight rabbits a 6.5% was observed. Weight gain from birth to weaning was lower in the comorbid group, reaching a litter weight of 4.5±0.13 kg in healthy normal weight does and 2.6±0.67 kg in comorbid does. The disturbances induced by the comorbidity were magnified in both behavioral and productive parameters.

Psoroptes ovis is a mite species that feeds on sheep, cattle, other ungulates, rabbits, and horses, which can develop into a severe exudative dermatitis known as psoroptic mange. The macrocyclic lactone (ML) family of acaricides are commonly used to control psoroptic mange. However, certain strains of cattle and sheep mites have developed resistance against MLs, which has led to reduced treatment efficacy and even treatment failure. Here we investigated the genetic basis of ML resistance in P. ovis mites collected from cattle across Belgium. We compared gene expression between susceptible and resistant mites before and after exposure to ivermectin and genetic diversity between a single susceptible and resistant populations. We generated chromosomal genome assemblies of P. ovis derived from sheep and cattle respectively and correlated genomic diversity of susceptible and resistant P. ovis populations sampled across Belgium. Gene expression data revealed constitutive over-expression of a cytochrome P450 monooxygenase (CYP) gene and two tandemly located UDP-glucuronosyltransferase (UGT) genes among others. On investigation of the genomic data, we observed copy number variation at both loci in population genomic data. The CYP gene is not amplified in the susceptible population but occurs in multiple copies in all resistant populations and is associated with a peak in F st between resistant and susceptible populations indicative of selection. By contrast, the two UGT genes are massively and tandemly amplified in all populations including the susceptible dataset with weaker F st between populations than the amplified CYP gene. Hence, distinct mechanisms of amplification and gene regulation are occurring at these putative resistance loci in P. ovis.

Psoroptes ovis, a common surface-living mite of domestic and wild animals worldwide, results in huge economic losses and serious welfare issues in the animal industry. P. ovis infestation rapidly causes massive eosinophil infiltration in skin lesions, and increasing research revealed that eosinophils might play an important role in the pathogenesis of P. ovis infestation. Intradermal injection of P. ovis antigen invoked massive eosinophil infiltration, suggesting that this mite should contain some relative molecules involved in eosinophil accumulation in the skin. However, these active molecules have not yet been identified. Herein, we identified macrophage migration inhibitor factor (MIF) in P. ovis (PsoMIF) using bioinformatics and molecular biology methods. Sequence analyses revealed that PsoMIF appeared with high similarity to the topology of monomer and trimer formation with host MIF (RMSD = 0.28 angstroms and 2.826 angstroms, respectively) but with differences in tautomerase and thiol-protein oxidoreductase active sites. Reverse transcription PCR analysis (qRT-PCR) results showed that PsoMIF was expressed throughout all the developmental stages of P. ovis, particularly with the highest expression in female mites. Immunolocalization revealed that MIF protein located in the ovary and oviduct of female mites and also localized throughout the stratum spinosum, stratum granulosum, and even basal layers of the epidermis in skin lesions caused by P. ovis. rPsoMIF significantly upregulated eosinophil-related gene expression both in vitro (PBMC: CCL5, CCL11; HaCaT: IL-3, IL-4, IL-5, CCL5, CCL11) and in vivo (rabbit: IL-5, CCL5, CCL11, P-selectin, ICAM-1). Moreover, rPsoMIF could induce cutaneous eosinophil accumulation in a rabbit model and increased the vascular permeability in a mouse model. Our findings indicated that PsoMIF served as one of the key molecules contributing to skin eosinophil accumulation in P. ovis infection of rabbits.