Failure of a single treatment with ivermectin to control sheep scab (Psoroptes ovis) on artificially infested sheep

Ivermectin at 200 micrograms/kg bodyweight given either as a single subcutaneous injection or as an oral drench failed to eradicate Psoroptes ovis from artificially infested sheep. The oral drench reduced the mite populations by 43 per cent within 24 hours but no further significant decline was recorded over 38 days. The subcutaneous injection reduced the mite populations by 90 per cent after 10 days but live P ovis were present on all the treated animals 84 days after treatment. The efficacy of treatment was less the higher the initial mite burden. The injection had no effect on clinical sheep scab, and the disease continued to progress despite the mite mortality.     

A simple pump-assisted method for collecting live,undamaged Psoroptes ovis from sheep using circulating saline

The availability of large numbers of undamaged sheep scab mites, Psoroptes ovis, would be beneficial for discovery screening and development trials. There are several reported procedures for removing scab mites from sheep but they have limitations. To overcome this, a simple but versatile method employing the use of pumped saline was developed to remove all stages of the P. ovis mite from sheep. The method takes no more than 2 min to remove mites from the selected site with relative ease and is not affected by the condition of the fleece or lesion. The number of mites removed with the new method was 5-10 times more than detected by visual examination. These mites were undamaged and survived off-host for up to 16 days. The robust, portable equipment is easy to use under field conditions, making this method suitable for use as a diagnostic tool for early detection and monitoring of scab mites thus providing opportunities for development of novel alternative control strategies.     

The construction of a cDNA expression library for the sheep scab mite Psoroptes ovis.

The need for alternative control strategies for sheep scab is critical. One approach is to develop vaccines based on 'concealed' antigens derived from Psoroptes ovis. This strategy requires the identification and characterisation of potential target antigens, which has been hampered by the problem of limited biological material for isolation of protein antigens. To aid the discovery of P. ovis antigens and to provide a resource for generating recombinant protein, we constructed a P. ovis cDNA expression library, using total RNA isolated from 250 mg of mixed-stage P. ovis and the Clontech SMART cDNA synthesis kit. The presence of P. ovis-specific sequences was confirmed using PCR amplification and sequencing of actin. The sequences of cDNA inserts from six random clones included one with high homology to the Dermatophagoides pteronyssinus (house dust mite) antigen p Dp15. This is a glutathione S-transferase known to be an important house dust mite antigen. We conclude that this library will be a useful tool for the identification of potential target antigens for the immunological control of P. ovis and to further our understanding of the pathology of sheep scab.     

The rate of spread of sheep scab within small groups of Merino and Dorper sheep

A single Merino sheep, artificially infested with the sheep scab mite, Psoroptes ovis, and a similarly infested Dorper sheep were placed with 9 uninfested Merino or 9 uninfested Dorper sheep respectively during winter and the rate of spread of infestation on the uninfested sheep observed. The same procedure was repeated in summer. It took 14 and 8 weeks respectively in winter before all sheep in the 2 groups displayed lesions of sheep scab, whereas in summer it took 10 and 12 weeks before all sheep had lesions.     

The proteinases of Psoroptes ovis, the sheep scab mite--their diversity and substrate specificity

The sheep scab mite, Psoroptes ovis, causes severe dermatitis in infected sheep with severe welfare and production implications. The dermatitis has the characteristics of an immediate hypersensitivity type reaction which, by analogy to other mite species, including the house dust mites (Dermatophagoides spp.), is likely to be invoked by a variety of allergens including mite-derived proteinases. Here, the proteinases in P. ovis extracts were characterised using substrate gel analysis, inhibitor sensitivity and their ability to degrade a variety of potential natural protein substrates. These analyses showed that mites contain several proteinases which could be differentiated on the basis of molecular size and inhibitor sensitivity with cysteine, metalloproteinases and aspartyl proteinases predominating. These proteinases degraded collagen and fibronectin, possibly indicative of a role in lesion initiation, they degraded several blood proteins, a property which may aid mite feeding and they degraded immunoglobulin G, possibly aiding immuno-evasion. Because proteinases, particularly the cysteine class, are demonstrably allergenic in other mite infestations, these proteinases clearly merit further immunological and biochemical definition.     

Immune responses to Staphylococcus aureus and Psoroptes ovis in sheep infected with P. ovis--the sheep scab mite

In sheep, lesions caused by Psoroptes ovis, the sheep scab mite, may become colonized by Staphylococcus aureus. The present study compares clinical signs, lesional area and the immune response to P. ovis and S. aureus in P. ovis-infested sheep with and without secondary S. aureus infection. No differences were detected in the clinical signs or lesional areas in the S. aureus-positive and -negative sheep. However, 6 weeks after infestation an IgG but not IgE isotype antibody response to S. aureus was detected in the S. aureus-positive but not the S. aureus-negative group of sheep. This response targeted S. aureus antigens with molecular weights of approximately 36, 38, 50 and 65 kDa. In addition, 6 weeks after infestation an IgE response to P. ovis was detected in the S. aureus-positive but not the S. aureus-negative group of sheep.     

The efficacy of showers for control of ectoparasites of sheep.

Ectoparasites of sheep in Britain include the tick Ixodes ricinus, the blowfly, Lucilia sericata, the ked, Melophagus ovinus and the lice, Damalinia ovis and Linognathus ovillus. The most important ectoparasite, however, is the mite Psoroptes communis ovis which causes sheep scab. This notifiable disease was eradicated from Britain in 1952 but was reported again in 1973. The control of ectoparasites depends largely on the efficient application of insecticide to the fleece and skin. Plunge dipping in a bath is generally regarded as the most efficient method and is compulsory for the control of sheep scab but it is not always the most convenient. After the eradication of sheep scab in 1952 plunge dipping was no longer compulsory and jetting, spraying and showering techniques were then employed for the other ectoparasites.     

Lipid ingestion from sheep epidermis by Psoroptes ovis (Acari: Psoroptidae)

Skin biopsies from three groups of sheep infested with Psoroptes ovis were cryofixed in liquid nitrogen to preserve outer epidermis with its lipid. From one group of five sheep (Group A), biopsies were taken from relatively healthy skin near the edge of scab lesions where mites had congregated. Frozen vertical sections from these biopsies were stained with Oil Red O and haematoxylin before mounting. Red lipid globules were plentiful within the body cavity of sectioned mites, but ingested lipid could not be distinguished from endogenous mite body stores by this technique. In a second group of five sheep (Group B), enclosed lumbar skin areas were coloured red with the lipophilic stains Oil Red O or Sudan IV. These enclosed coloured skin areas were inoculated with P. ovis and sampled by skin biopsy 1 or 4 days later. Cryofixed biopsies were cut into vertical frozen sections and mounted without staining for examination. Red-coloured lipid within mites, matching red-coloured lipid on outer epidermis, was evidence for the epidermal origin of P. ovis ingesta in the early stages of an infestation. From two other sheep (Group C), cryofixed biopsies were examined by scanning electron microscope and mites were seen with mouthparts embedded in abraded outer epidermis, but the precise depth of epidermal penetration was not determined. A light microscope survey of 3198 frozen vertical skin sections from the first 10 sheep (Groups A and B) showed that inner stratum corneum of the epidermis was the deepest penetration recorded for gnathosomes of P. ovis cryofixed in situ by liquid nitrogen. No structure of P. ovis was identified in dermal tissues.     

不同温湿度和培养环境对离体绵羊痒螨生存活力的影响

目的 测不同温度及培养环境对离体绵羊痒螨生存能力的影响,进一步了解痒螨的传播机制,为制定防治瘁螨的有效措施提供理论基础。方法 置室内外自然温度和实验室恒温条件,以及5种相对湿度：50、60、70、80及90％RH,培养观察绵羊痒螨不同虫态（卵、幼虫、幼虫静止期、若虫、若虫静止期、雌虫、雄虫）的生存发育情况;同时观察痒螨在水、痂皮屑、羊毛、羊粪、土壤、皮组织等6种条件下的生存活力。结果 痒螨的最适生长发育温度及繁殖期在21℃-28℃范围内,随着温度的升高,痒螨存活时间缩短,绵羊痒螨对4℃低温有一定耐受力,但对35℃以上的高温耐受力较弱。在同一温度下,相对湿度越高,各虫态存活时间越长。结论 体外观察实验表明痒螨对外界环境有较强的适应能力,在缺乏营养的条件下仍能存活较长一段时间。     

Expression and characterisation of a Psoroptes ovis glutathione S-transferase

The astigmatid mite Psoroptes ovis is the causative agent of sheep scab, a highly contagious parasitic disease of sheep. Infection causes severe allergic dermatitis, resulting in damage to the fleece and hide, loss of condition and occasional mortality. Interest in the P. ovis allergens led us to characterise a glutathione S-transferase (GST) which displays homology to GST allergens isolated from the house dust mite, Dermatophagoides pteronyssinus and the cockroach, Blatella germanica. A cDNA encoding a mu-class GST from P. ovis was expressed in Escherichia coli and the recombinant protein purified for biochemical analysis. SDS-PAGE analysis indicated that the purified product was homogeneous and had an apparent molecular weight of 30 kDa. The recombinant GST (rGST) is active towards the substrate 1-chloro-2,4-dinitrobenzene (CDNB), whereas 1,2-dichloro-4-nitrobenzene (DCNB) is a poor substrate. The recombinant protein was also tested for recognition by IgE and IgG antibodies in serum from P. ovis na?ve and P. ovis infested sheep. Neither IgE nor IgG antibodies were detected to the rGST. Prausnitz--Küstner testing with rGST did not provoke a characteristic weal and flare response. Biopsies collected at the PK test sites were stained for eosinophils, neutrophils, mast cells and basophils. Neutrophil, mast cell and basophil counts were not significantly different to the controls. Eosinophil numbers were significantly higher than controls, but were not due to an IgE response.     

The effects of temperature and humidity on the off-host survival of Psoroptes ovis and Psoroptes cuniculi.

The effects of both temperature and humidity on the survival of the mites Psoroptes ovis and Psoroptes cuniculi were considered in laboratory assays. When P. ovis and P. cuniculi were maintained at 95% humidity, maximum survival decreased linearly with increasing temperature, from approximately 15 days at 9 degrees C to 5 days at 30 degrees C. There was no significant difference between P. ovis and P. cuniculi in the effects of temperature on maximum survival. Adult male P. ovis and P. cuniculi had lower mean maximum survival than any of the other life cycle stages. There was a small but significant effect of humidity on survival for P. cuniculi; LT50 values were greater at 75-85% r.h. than at 55-65% r.h. The influence of off-host survival and the infestation of naive sheep from mites in the environment on the epidemiology of sheep scab are discussed.     

Approaches to vaccines for Psoroptes ovis (sheep scab)

A brief outline of the history of sheep scab in the UK is presented together with the current chemical methods used for its control and problems associated with these. Possible alternative approaches to control are discussed, as are selected aspects of the physiology of Psoroptes ovis and the pathogenesis of the scab lesion from the perspective of control through immunisation. Evidence is provided that immunity to the disease can indeed be acquired, both naturally after a previous infection and following inoculation of mite proteins in adjuvant. These results support the view that control by vaccination may be possible, although little is known to date about the antigens involved or the mechanism of protection.     

The effect of immunosuppression with cyclosporin A on the development of sheep scab

The present study confirms that following infection with the ectoparasitic sheep scab mite, Psoroptes ovis, there is a rapid (within 24 h) inflammatory influx of eosinophils and apoptosis of the keratinocytes at the site of infection. In order to investigate whether these inflammatory reactions are important in the maintenance of mite infection, a group of animals were treated daily after the establishment of infection with the potent anti-inflammatory drug, Cyclosporin A. The course of infection was monitored by determining the lesion area and mite numbers, systemic antibody and blood eosinophils, as well as the inflammatory cells and T cell sub-populations within the lesion throughout the 6-week duration of the experiment. These parameters were compared with those in a similar infected control (non-treated) group. In control infected animals, the lesion area and mite numbers increased steadily throughout the 6-week period. In contrast, lesion area and mite numbers were severely depressed in the group which received Cyclosporin A. Local and systemic eosinophils, and systemic antibody were also significantly reduced in the drug treated animals, compared to controls. Surprisingly however, the number of lesional pan T cells, T helper cells, gammadeltaT cells and dendritic cells in Cyclosporin A treated animals were either the same, or significantly (P < 0.05) enhanced when compared to the control infected animals at the termination of the experiment. The results will be discussed in terms of the role of the dermal inflammatory response in the establishment and maintenance of the sheep scab mite.     

5%AVM预防绵羊疥癣病的长效试验

目的检测5%AVM对绵羊疥癣病的预防效果。方法通过痒螨检查确定试验羊,设Ⅰ、Ⅱ、Ⅲ5%AVM三个剂量组,Ⅳ为1%阿维菌素注射液组,Ⅴ为螨净组,Ⅵ为空白对照组,每个组7只试验羊;Ⅶ为传染源组。按羊的体重皮下注射或药浴,观察近90d。结果Ⅰ组61d发病;Ⅱ、Ⅲ组92d发病;Ⅳ组35d发病;Ⅴ组没有发病;Ⅵ组21d发病。结论5%AVM有效预防绵羊疥癣病近90d。     

Diagnosis of psoroptic sheep scab with an improved enzyme-linked immunosorbent assay

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of specific antibodies against crude Psoroptes antigen. The diagnostic sensitivity was 93.7% in 191 sheep with clinical signs associated with mange. These animals originated from 29 flocks in which psoroptic mites were detected. All of 59 sheep infested with Psoroptes ovis were seropositive. Additionally, in 49% of 70 clinically unaffected sheep originating from P. ovis-infested flocks, specific antibodies could be detected, suggesting that asymptomatic infestations can be diagnosed by serology. The specificity of the ELISA was 96.5% as determined with 254 sheep originating from 44 flocks without clinical mange. Cross-reactivity in a low range was detected with selected sera of sheep with clinical chorioptic or forage mite infestations. Four sheep seroconverted 2 weeks after experimental P. ovis infestation, i.e. 2 weeks before clinical signs became obvious. After successful doramectin treatment of 14 sheep with naturally acquired P. ovis infestation, the ELISA values declined slowly but remained positive in seven cases beyond 17 weeks.     

Outbreak of sheep scab on two adjacent common alpine pastures in the canton of Schwyz (Switzerland)

Sheep scab, which is caused by the mite Psoroptes ovis, is a notifiable disease in Switzerland. In the framework of an epidemiological study in the canton of Schwyz, a sheep scab outbreak on 2 adjacent alpine pastures with 62 flocks (1770 sheep) was followed up clinically, parasitologically and serologically. No mass treatment of the flocks with organophosphates or avermectins had been undertaken before the flocks were taken up to the alpine pastures, but they were treated according to their serological status. Supervised treatment of seropositive judged flocks (at least one seropositive or at least 2 serologically equivocal animals per flock) with Doramectin was undertaken, whilst seronegative judged flocks remained untreated. Sheep returned from the summer pastures in early September 2001 and within three months scab infestation was diagnosed in 53 flocks (85.5%). These infested flocks were scattered all over the canton. Furthermore, the transfer of sheep from 6 infested flocks resulted in the transmission of the disease to 10 new flocks. Infested flocks were kept in quarantine and sanitized. The complexity of the traditional use of shared alpine pastures and the intense, uncontrolled trading with animals render the control of sheep scab difficult.     

Clinical development and serological antibody responses in sheep and rabbits experimentally infested with Psoroptes ovis and Psoroptes cuniculi

Psoroptes ovis of sheep origin, and Psoroptes cuniculi of rabbit origin were used in experimental infestations. In experiment I, groups of four rabbits and four sheep were infested with 50-100 mites of each isolate on the skin of the back (skin infestation, SI) or in the external auditory canal (aural infestation, AI). In rabbits, SI and AI with P. cuniculi and AI with P. ovis induced in all animals typical ear lesions and pronounced antibody reactions to P. cuniculi antigens in ELISA. After SI of rabbits with P. ovis no clinical signs were detected, no mites could be reisolated and no specific antibodies were detected. In sheep, P. ovis SI induced mange whereas AI did not induce typical clinical signs and mites could not be reisolated. In both these animal groups, ELISA revealed pronounced and comparable specific antibody reactions. After SI and AI with P. cuniculi no clinical symptoms were observed and no mites could be reisolated. Nevertheless, low levels of specific antibody were detected. In experiment II, clinical progression and antibody reactions to P. ovis SI in naive sheep were compared with sheep previously exposed to P. ovis or P. cuniculi. In both pre-exposed groups of animals, clinical signs appeared within 2 days after challenge infestation and three days earlier than in primarily infested sheep. Subsequently, no obvious difference in the clinical progression was observed between the three groups of animals. The results of this study document antigenetic crossreactivity of the two morphologically and genetically distinguishable Psoroptes species but differences in their biological behaviour and virulence which both are of epidemiological and taxonomic relevance.     

The in vitro response of sheep scab mites to pyrethroid insecticides.

The response of sheep scab mites to pyrethroid insecticides and organophosphate compounds was studied in vitro with the objective of finding a simple test for detecting insecticide resistance in scab mites. Psoroptes cuniculi from rabbits or P. ovis from sheep were enclosed in small 'tea bags' made from heat sealable paper prior to dipping in insecticide. Mites failed to die 24 h after a 1 min dip in working concentrations of insecticidal sheep dips. With flumethrin a variety of different conditions were tested but most failed to improve the efficacy of flumethrin. It is suggested with freshly collected mites pyrethroids may be taken up by ingestion.     

Treatment and control of sheep scab (Psoroptes ovis) with ivermectin under field conditions in South Africa.

Four trials including 11,266 sheep were conducted in South Africa to evaluate the efficacy of the systemic parasiticide ivermectin against field outbreaks of sheep scab (Psoroptes ovis) when two doses of approximately 200 micrograms/kg were administered subcutaneously seven days apart (days 0 and 7). As sheep scab is a notifiable disease in South Africa, it was not possible to include an untreated control group. The prevalence of clinically affected animals in the four treated flocks varied from 0.4 per cent to 99 per cent before the two treatments. After the treatments, there were no signs of active clinical infection in any of the sheep between days 28 and 30, or at subsequent examinations. P ovis mites were recovered from scrapings from 114 of 127 indicator sheep before the treatment but no mites were recovered from them between days 28 and 30 or 42 and 58 after the treatments.     

Ultrastructure of the alimentary canal of the sheep scab mite, Psoroptes ovis (Acari: Psoroptidae).

The mite Psoroptes ovis causes sheep scab disease in flocks throughout much of the world. A serious impediment to the development of novel control measures for this mite is our inability to produce in vitro colonies of the mite. Here, we describe the alimentary canal of the mite with the particular aim of determining what it feeds on, as part of a longer term goal, to develop in vitro culture techniques for P. ovis. The alimentary canal of P. ovis consists of a cuticle-lined foregut and hindgut separated by a microvilli-lined midgut. The foregut is divided into a pre-oral cavity and a muscular pharynx and oesophagus. The midgut consists of three ultrastructurally discrete areas: a stomach with bi-lobed ventriculi, a colon and a post-colon. The stomach is composed of two cell types. The most common cells (Type 1) are either cuboidal or squamous depending on the degree of gut distension and possess short microvilli, a single basally located nucleus, extensive rough endoplasmic reticulum and other components suggesting active secretion. The less common cells (Type 2), possess an extensive apical network of tubules and basally food vacuoles suggesting intracellular digestion. These cells extend into the gut lumen, become free-floating and degenerate. The colon and post-colon are composed of cells similar to Type 1 stomach cells but the post-colon epithelium possesses significantly longer microvilli. Cells from these areas have not been observed to leave the surrounding epithelium and enter the gut lumen, but it is suggested that significant absorption occurs in these two areas. Faecal pellets, often containing a significant number of bacteria, leave the digestive system through the cuticle-lined anal atrium.