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.     

Host preference of the sheep scab mite,Psoroptes ovis

Sheep scab mites, Psoroptes ovis, collected from a Merino donor sheep, were used to infest Merino and Dorper sheep, and Angora and Boer goats. Mites were placed on the sheep on 1 or 2 occasions and on 5 occasions on the goats. All the animals were examined at regular intervals for the presence of scab lesions and living mites. Both sheep breeds developed lesions, but those on the Merino sheep were always larger than those on the Dorper sheep at the same intervals after infestation. None of the goats developed lesions or showed signs of irritation, or harboured any mites.     

Temporal pattern of isotype-specific antibody responses in primary and challenge infestations of sheep with Psoroptes ovis--the sheep scab mite

In sheep Psoroptes ovis provokes an allergic dermatitis with significant P. ovis antigen-specific IgE responses. The kinetics of the IgE response to primary and challenge infestations of P. ovis were reported earlier [Parasite Immunol. 22 (2000) 407]. The present study examines IgG, IgM and IgA responses to primary and challenge infestations of P. ovis and the profile of antigens/allergens reacting with IgG and IgE antibodies. Antigen-specific enzyme-linked immunosorbent assays (ELISAs) demonstrate that primary infestations elicited significant increases in levels of IgG and IgM but not IgA antibodies. IgG and IgM responses to primary and challenge infestations were not significantly different. Western blots of reduced P. ovis proteins indicate that IgG antibodies reacted with five major antigens following primary infestation and only three of these after challenge infestation. IgE antibodies bound to three major and five minor allergens after primary infestation and two additional minor allergens after challenge infestation. Immunodominant antigens >100 and <15 kDa and allergens >100 kDa were most consistent in stimulating substantial IgG and IgE antibody responses, respectively. These antigens/allergens may be exploited in immunodiagnosis and modulation of the host immune response.     

On-host ecology and off-host survival of the sheep scab mite Psoroptes ovis

These studies were conducted to investigate the possible role of certain aspects of the on-host ecology and off-host survival of the sheep scab mite, Psoroptes ovis, in the dissemination of infestation. All developmental stages of the mite occurred in the fleece both proximally or distal to the skin of infested Merino and Dorper sheep. A larger proportion of mites was present in the fleece of Dorper sheep distal to the skin in the late afternoon and early morning than at other times during the day. Immature and adult mites readily transferred to tufts of wool or hair placed on infested sheep of both breeds. No mites could be found on wool or hair rubbed off onto tree trunks or branches or other structures in enclosures housing heavily infested sheep, nor could any mites be collected from the soil of these enclosures, whereas more than 80% of mites artificially seeded onto soil samples were recovered. The longest mean off-host survival times for larvae, nymphs, and male and ovigerous female mites were recorded at 10 degrees C, and were 9.25 days (RH = 90%), 15 days (RH = 33% and 75%), 10.5 days (RH = 75% and 90%) and 11.25 days (RH = 90%) respectively. Under natural climatic conditions ovigerous females in glass vials containing Merino wool survived for 17 days compared to 15 days for females in vials without wool; this difference was, however, not significant. The mean off-host pre-hatch period for eggs varied between 5.9 days (T = 25 degrees C and RH = 33%) and 22.1 days (T = 10 degrees C and RH = 75%), while the longest time individual eggs took to hatch at the latter temperature and RH was 31 days.     

The behaviour of sheep with sheep scab, Psoroptes ovis infestation.

Methods of behavioural data collection as applied to the study of sheep scab are described. The behaviour of a number of marked animals within an affected flock was recorded using an event recorder. Focal sampled data were analysed for duration and frequency of the observed behaviours. In addition, the flock was observed at intervals and the instantaneous behaviour of each animal recorded. These data were compared with similar observational data from the same flock recorded at intervals during post-treatment recovery. Sheep scab resulted in pathological behaviours of rubbing, scratching and biting at the lesion, and these resulted in interruption of the normal behaviours, grazing, cudding and idling, but did not result in reduced levels of these behaviours. Infested sheep showed stereotypic mouthing behaviour, initiated by rubbing or scratching, or in some cases without any external stimulus. No stereotypic behaviour was seen in animals after treatment. The implications of the behavioural data for the welfare of the sheep is discussed.     

Haematological and immunological response of unrestrained cattle to Psoroptes ovis, the sheep scab mite

Cows were infected twice with 600 and 500 nymphs and adults of a bovine strain of Psoroptes ovis with a nine-week interval. The haematological response and the non-specific mitogen- and antigen-induced responsiveness of the peripheral blood lymphocytes of the animals was followed. Dermal reactivity to P ovis antigen injection was studied five weeks after reinfection. After the first infection with 600 mites none of the infected animals developed clinical psoroptic mange but a leucocytosis developed, contributed to primarily by an eosinophilia and by a slight lymphocytosis. Antigen-induced lymphocyte blastogenesis was used to measure the antigen-sensitive cell population in peripheral blood and this population showed a maximum increase 10 days after infection; however, antigen-sensitive cells remained above normal levels until reinfection. Upon challenge infection with 500 mites the infected animals showed an immediate hypersensitivity type reaction with a marked pruritus, scratching and exudation. Thereafter the lesions healed rapidly and none of the animals developed clinical mange. This clinical reaction was accompanied by a secondary eosinophilia but no change was apparent in the other blood elements. A marked increase in the blastogenic response of the peripheral blood lymphocytes was also apparent and this peaked three weeks after challenge. Following the intradermal injection of P ovis antigen there was an immediate swelling of the injection site in all infected and control animals and skin thickness was maximal one hour after injection. Thereafter there was a clear distinction in dermal reactions between P ovis infected and control animals; after 48 hours reactions were not seen in the control animals while marked dermal reactions were still present in the P ovis infected group.     

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.     

Enzyme-linked immunosorbent assay for the detection of antibodies to the sheep scab mite Psoroptes ovis

An enzyme-linked immunosorbent assay has been shown to detect antibodies in the circulation of sheep infected with Psoroptes ovis. Strong positive reactions were obtained from 19 sheep with four-month-old infections. No cross reaction was observed with sera obtained from sheep infected with either Fasciola hepatica, Nematodirus battus, Ostertagia circumcincta or Damalinia ovis.     

Localisation and characterisation of ovine immunglobulin within the sheep scab mite, Psoroptes ovis

Ovine IgG was detected in homogenates of repeatedly washed Psoroptes ovis. Some of the immunoglobulin in the homogenates was fragmented although the host IgG present in mite washings was largely intact. The host immunoglobulin was immuno-localised to the surface or cytoplasm of the gut cells of feeding stages of freshly harvested P. ovis examined by cryosectioning. A similar distribution of rabbit IgG was detected in P. cuniculi. The IgG demonstrated in the mite gut represented partially digested as well as intact immunoglobulin. The presence of intact host immunoglobulin suggests that P. ovis may be susceptible to vaccination by the gut antigen approach, a method used successfully for blood-feeding ectoparasites like Boophilus microplus.     

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.     

Control of the sheep scab mite Psoroptes ovis in vivo and in vitro using fungal pathogens

As part of a research programme designed to identify biological agents for the control of sheep scab, the pathogenicity of the fungus Metarhizium anisopliae to Psoroptes mites in the presence of sheepskin and wool was examined in the laboratory. No inhibitory effects of skin and wool were observed and high levels of infection were recorded. Subsequently the pathogenicity of formulations of both M. anisopliae and Beauveria bassiana to Psoroptes ovis was studied in vivo. For this, 36 batches of 20 adult female Psoroptes mites were confined in 25 mm diameter chambers which were attached to the backs of 6 scab-naive sheep. In some treatments, mites were exposed to the fungal pathogens for 48 h in vitro prior to being placed on the host, while other treatments involved mites with no prior exposure placed directly onto the skin of a host treated with a fungal pathogen. After 48 h on the host, mites were removed, incubated individually and all fungal infections were recorded. Fungal infection was observed in all treatments, except untreated controls. However, B. bassiana infected a significantly greater number of mites than M. anisopliae with all the formulations examined. Infection rates were highest when mites were exposed to dry conidia (>90%) and lowest with M. anisopliae in diatomaceous earth. Overall, the infection rate was not affected by whether or not the mites were given prior exposure to the conidia, before being placed on the sheep. The results demonstrate that Psoroptes mites can become infected by entomopathogenic fungi on the skin of sheep and provides a first demonstration of the potential of this technology for the control of sheep scab.     

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.     

Effects of the scab mite Psoroptes ovis on the haematology and live mass of Merino and Dorper sheep

Five Merino and five Dorper sheep were artificially infested with the sheep scab mite Psoroptes ovis and the effect of infestation on their haematology, serum protein levels and live mass recorded for a period of 14 weeks. The reaction of the Merino sheep to infestation was more severe than that of the Dorper sheep. Haematological values fluctuated within the normal range during the assessment period. The mean haemoglobin concentration of the Merino sheep declined until antiparastic treatment was administered 10 weeks after infestation, after which it gradually increased. The lymphocyte counts of both breeds of sheep declined from 2 weeks to 10 weeks post-infestation, but increased after treatment, while the highest eosinophil counts were recorded in the Merino sheep at the height of the acute disease 8-10 weeks post-infestation. Serum albumin values for both breeds and serum globulin values for the Merino sheep were higher than normal during the entire 14-week observation period. A decrease in serum albumin and an increase in serum globulin concentration occurred at the height of infestation in both breeds. The mean live mass of a second group of five infested Merino sheep decreased by 6.4 kg over a 16-week period compared to a gain of 4.56 kg for five infested Dorper sheep.     

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.     

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.     

Some observations on the biology and control of the sheep scab mite Psoroptes ovis (Hering) in Britain

Sheep scab was eradicated from Britain in 1952 but reappeared in 1973. The host specificity of the sheep scab mite is discussed; sheep are the only reservoir of the disease in Britain. Control is by single dipping in the approved acaricides, HCH, diazinon or propetamphos. Acaricides are approved only if the minimum use rate (Maintenance level) will give at least 4 weeks' protection. The method of testing is described. Traditionally, compulsory dipping has been in the winter when the disease is most active. Although the mites were originally thought to enter a latent phase during the summer, new observations indicate that there is little difference between their summer and winter behaviour. In Britain, dipping is now compulsory in both the summer and winter.     

Characterisation of lesional infiltrates of dendritic cells and T cell subtypes during primary infestation of sheep with Psoroptes ovis, the sheep scab mite

Earlier studies of cattle and sheep have demonstrated that Psoroptes ovis infestations provoke an intense immunoinflammatory response dominated by eosinophils accompanied by a substantial infiltrate of lymphocytes. However, the kinetics of the lymphocyte response and the subtypes involved have not been characterised. We employed two groups of sheep to investigate the early (1-21 days) and later (21-63 days) infiltration of lymphocyte subpopulations and dendritic cells in primary infestations of sheep with P. ovis. Immunohistochemistry indicated that by 4 days after infestation numbers of CD4+ and CD45RA+ cells in lesional skin had increased significantly (P<0.03 and P<0.005, respectively) and that a significant increase in gammadelta T cells and dendritic cells (CD1b+) had occurred by 8 days (P<0.02 and P<0.01, respectively). Numbers of lymphocyte and dendritic cells declined from 49 to 63 days after infestation. Our observations suggest that mite-derived products exert a profound influence on the early recruitment of lymphocytes that may significantly influence the genesis of the adaptive immune response.     

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.