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.     

Dermatopathology and immunity in experimental Psoroptes ovis (Acari:Psoroptidae) infestation of naive and previously exposed Hereford cattle

Naive and previously exposed young Hereford cattle were infested with Psoroptes ovis and maintained in stanchions for 7 weeks. Skin biopsy samples were examined at 1, 2, 5, and 7 weeks after infestation was induced, compared with those from noninfested control calves, and correlated with changes in the mite populations and development of dermatitis. Gross and microscopic appearance of lesions were qualitatively similar in both groups of infested calves. The lesions occurred early in the previously exposed calves, but progressed slowly, whereas lesions in the naive calves appeared later, but were rapidly progressive. The basic histopathologic pattern was chronic exudative superficial perivascular dermatitis compatible with allergy. The mite populations on the naive calves revealed exponential growth and high fecundity, and reached high population densities, whereas mite populations on previously exposed calves had low growth rates and low fecundity and reached population densities 100 to 1,000 times less than those on naive calves. Seemingly, there was a hypersensitivity to mite antigens. The hypersensitivity response may have been a mechanism by which humoral immunity was delivered to the mite microenvironment and may have contributed to acquired resistance by controlling mite populations. The mechanism of population control seems to be related partially to decreased fecundity of female P ovis.     

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.     

The pathology of Psoroptes ovis infestation in cattle with a special emphasis on breed difference.

In cattle, infestations with P. ovis are responsible for a severe dermatitis. The disease is very common in some breeds of beef cattle whereas dairy cattle such as Frisian Holstein are considered as resistant. In order to investigate the factors responsible for this marked breed susceptibility, the immune response to P. ovis under experimental and field conditions has been studied using serology, lymphocyte transformation assay, intradermal tests and passive cutaneous anaphylaxis in susceptible Belgian White and Blue (BWB) and resistant Frisian Holstein (FH) cattle. The results of published studies are reviewed in this paper; original data on specific hypersensitivity reactions to P. ovisare also presented. The use of a sandwich ELISA allowed the detection of specific antibodies as early as 7 days post-infestation and very high titres were observed. There was a clear positive correlation between the antibody titre and the extent of lesions. This antibody response was associated with a marked in vitro lymphocyte proliferation but there was no difference between susceptible and resistant cattle. The passive cutaneous anaphylaxis test revealed the presence of specific Ig E in actively infested animals irrespective of the breed. Specific Ig E were detected fairly late during the course of an experimental infestation with P. ovis. The intradermal injection of P. cuniculi antigenic extract induced an immediate reaction in all animals including the controls; in contrast, a delayed hypersensitivity reaction was observed in infected BWB animals only. It is concluded that immunological as well as non-immunological factors may be responsible for the breed related susceptibility or resistance to P. ovis.     

Development of serum antibody activity as determined by enzyme-linked immunosorbent assay to Psoroptes ovis (Acarina:Psoroptidae) antigens in cattle infested with P. ovis

Calves infested with Psoroptes ovis (Hering, 1838) for the first time (naive) or previously infested calves were examined for serum antibody activity to P. ovis (obtained from rabbits) antigens by enzyme-linked immunosorbent assay (ELISA) to determine the temporal appearance of specific serum antibody activity. The development of the serum antibody activity to P. ovis antigens was then correlated with the development of lesions (% scab) and with changes in the number of P. ovis. The serum antibody activity [ELISA OD414 value (EODV) greater than 0.290] to P. ovis antigens in naive calves, in most cases, is first detected at the same time or slightly after the detection of mites and mite-caused lesions; and the development of specific serum antibody activity paralleled the increase in the P. ovis population and the percentage scab until 7 weeks post-infestation (PI). In previously infested calves, if serum antibody activity to P. ovis antigens was not already present from the previous infestation, specific serum antibody activity was detected at the same time and developed in a similar manner as in the naive calves. The serum antibody activity to P. ovis antigens could be detected after calves were relieved of their P. ovis burdens by pesticide treatment or after the P. ovis population began to decline when the calves were allowed to groom themselves. Serum antibody activity to P. ovis antigens was not detected in any of the control calves during the test period.     

Failure to protect goats following vaccination with soluble proteins of Sarcoptes scabiei: evidence for a role for IgE antibody in protection

Developing an anti-scabies vaccine is thought to be a feasible alternative to chemical control, since animals which have recovered from sarcoptic mange become resistant against mite reinfestation. The purpose of this study was to evaluate the protective value of immune responses developed in animals after immunisation with soluble mite proteins. Soluble proteins from Sarcoptes scabiei were extracted then subjected to ion exchange chromatography, and proteins from the column were eluted step-wise with 0%, 10%, 25% and 50% of 1 M solution of NaCl in a Tris buffer. Each protein fraction was concentrated and dialysed against PBS. To evaluate the immunogenicity of the fractions, 36 goats were allocated into six groups, group1 goats were unvaccinated, group 2 were vaccinated with intact soluble mite proteins, and groups 3-6 were vaccinated respectively with the fractionated proteins. Vaccinations were conducted four times with 1 mg protein/dose and 4-week intervals between vaccinations. One week after the last vaccination, all goats were challenged with approximately 2000 live mites on the auricles and infestations were allowed to progress for 6 weeks. The severity of lesions caused by the infestation was assessed throughout the study. The challenge caused mange or encrustation dermatitis in all animals and no differences in severity of lesions were observed between vaccinated and unvaccinated control goats. Vaccination with each fraction of the mite proteins invoked high levels of scabies-specific IgG in the serum of all animals but failed to induce specific IgE as determined by Elisa. In contrast, goats challenged experimentally with a primary or repeated mite challenge developed strong serum IgE and IgG antibody responses to Sarcoptes antigens. The latter animals were shown in a previous study to be resistant to reinfestation. The lack of immune protection in the vaccinated animals may be attributed to the absence of protective levels of IgE antibody, and the present findings indicate that allergens and IgE antibody is important in immunity to S. scabiei infection.     

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.     

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.     

Simulation analysis of the population dynamics of the mite, Psoroptes ovis, infesting sheep.

The pattern of population growth of the ectoparasitic mite, Psoroptes ovis (Acari: Psoroptidae), on its ovine host is considered through the development of a Leslie matrix-based, simulation model. The model is parameterised using experimental data in conjunction with reanalysis of published data. The model shows that on sheep P. ovis populations grow at a rate of approximately 11% per day and the population doubles every 6.3 days. Additional rates of adult mortality, in excess of 50% per day, need to be imposed to prevent population growth. The predictions of the model are tested by comparison of the expected numbers of mites with the numbers recorded in lesions either on naturally infested sheep where the date of infestation can be estimated or on one artificially infested animal, where the initial number of mites and date of infestation are known precisely. In both cases the observed number of mites in lesions relate closely to the numbers expected from the simulations. The model simulations do not support the concept of a 'lag' phase as distinct from the 'growth' phase in the changing pattern of mite abundance on an infested sheep and suggests that the observed pattern of growth is a natural function of an exponential increase in numbers. The development of such models and their use in explaining the demographic processes which drive mite population dynamics are discussed.     

Bone marrow response in cattle with chronic dermatitis caused by Psoroptes ovis

Six Hereford heifer calves were infested with Psoroptes ovis and compared to six uninfested control calves. Infested calves developed severe exudative dermatitis which covered 100% of the body surface by 7 weeks after infestation. They developed moderate nonregenerative anemia, decreased numbers of neutrophils and lymphocytes, and increased plasma protein and fibrinogen concentrations. Sternal marrow core biopsies taken 7 weeks after infestation were 75 to 90% cellular, while cores from uninfested calves were 10 to 20% cellular. Comparison of myelograms between the two groups of calves revealed increased M:E ratio, myelocyte and metamyelocyte hyperplasia with depletion of mature neutrophils and eosinophils in the P. ovis-infested calves. It was concluded that P. ovis-infested calves develop myeloid hyperplasia and that mange-related neutropenia is not due to bone marrow suppression by a mite or scab-associated toxin.     

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.     

Effects of Psoroptes ovis (Acarina:Psoroptidae) on certain biochemical constituents of cattle serum

Sera from Psoroptes ovis-infested calves were examined for 17 selected biochemical constituents to determine if P. ovis caused changes in any of these constituents. In order to recognize a scabies effect, there had to be a statistically significant (P less than or equal to 0.10) scabies exposure X time interaction and the changes in the biochemical constituents should have been correlated with the changes in the mite infestation. Five serum biochemical constituents fulfilled these criteria. Total protein, alpha-, beta-, and gamma-globulin were increased and total cholesterol was decreased at the peak of the scabies infestation. Precipitating antibodies to a P. ovis antigen were detected by immunodiffusion in some of the stored sera of the infested calves just after the peak of the infestation. Total protein, beta-, and gamma-globulin values were probably increased as a result of antibody production; alpha-globulin values were probably increased as a result of inflammation. As the scabies infestation declined, the mean values of all the above constituents from the infested calves, except cholesterol, began declining. Decreases in total cholesterol occurred, but they could not be entirely attributed to scabies. Examination of the serum biochemical constituents from a heavily infested calf that died during the test suggested that, in addition to antibody production and inflammation, kidney and liver damage may have occurred.     

Oestrus ovis (Diptera: Oestridae): sheep humoral immune response to purified excreted/secreted salivary gland 28 kDa antigen complex from second and third instar larvae

The potential of Oestrus ovis larvae excretory-secretory products (ESP), crude extract, salivary gland contents (SGc) and digestive tube contents (DTc) proteins from O. ovis larvae as antigens in serological assay were analyzed and compared. This study demonstrated that excretory-secretory proteins were produced mainly by digestive tube and salivary gland. The main antigenic component of ESP was a 28kDa protein complex (pc28) synthesized by the salivary gland ring. After purification, its diagnostic value was estimated by an ELISA using sheep sera sampled in summer (n=30) and winter (n=69) in order to investigate the relative importance of wintering hypobiotic first instar larvae on animal responsiveness. ELISA test based on pc28 antigen was compared to a reference ELISA test based on second instar larvae crude extract. After comparison it appeared that the pc28 test was much more specific (87%) and more sensitive (90%) in summer. Anti-pc28 antibody titers were significantly higher in summer than in winter demonstrating the impact of summer developing larvae on the humoral systemic immune response. This data was correlated to the maturation of larvae in summer as the number of larvae at necropsy was significantly higher in winter, when the larval population is only represented by first instar larvae, than in summer. No correlation was observed between the mean antibody titers and the number of larvae.     

Lymphocyte responsiveness in cattle previously infested and uninfested with Hypoderma lineatum (de Vill.) and H. bovis (L.) (Diptera: Oestridae)

Cattle exposed to their third consecutive warble (Hypoderma lineatum and H. bovis) infestation had significantly reduced apparent and accumulative grub populations and produced significantly fewer grubs than animals exposed to their first infestation. These resistant animals had enhanced reactivity to mitogens (concanavalin A (con A) and pokeweed) upon reinfestation and a strong antigen-specific response at 2 months post-infestation. Their responsiveness to con A and antigen was also enhanced at 7 months post-infestation when grubs appeared in the backs, and the response to antigen continued at 10 months post-infestation. Responsiveness to mitogens in previously uninfested animals was similar to the control response throughout the infestation apart from the suppression of the con A response at 7 months post-infestation. These animals finally responded to antigen at 10 months post-infestation. These results suggest that acquired resistance to hypodermosis has a cellular basis with participation of both B- and T-cell components.     

Overberg research projects. VI. The biology and control of Oestrus ovis in sheep in the winter rainfall areas of the southern Cape

Oestrus ovis was endemic on all the farms included in a survey conducted in the southern Cape, but each farm had its own unique seasonal pattern of infestation. Flock sheep were infested 10-12 months and tracers 5-9 months of the year. Sporadic infestations occurred in winter and spring, while peaks were reached in summer and autumn. Development of O. ovis larvae deposited in autumn was retarded for up to 5 1/2 months. Pupae of O. ovis formed from 27 April-9 August, with the exception of a single pupae formed on 29 June, failed to produce flies. Pupal periods ranged from 30 days in January to 80 days in June. Strategic anthelmintic treatments in May, August and November and a tactical treatment in March are recommended.     

Inter- and intra-specific variation within the genus Psoroptes (Acari: Psoroptidae).

In this paper the taxonomy and relative importance of various species within the genus Psoroptes are reviewed with respect to inter- and intra-specific variations in cross infestivity and host specificity, site of infestation and morphological characters. Intra-specific variation within populations of the sheep scab mite, Psoroptes ovis will then be discussed critically with respect to virulence (the relative growth of the scab lesion and mite burdens), epidemiology, susceptibility to the systemic acaricide, ivermectin, and resistance to synthetic pyrethroid and organophosphate acaricides. Finally, it is suggested that the ear mite, P. cuniculi, and the sheep scab mite, P. ovis are variants of the same species. Information was collected from published literature and from the results of unpublished studies carried out at the VLA, Weybridge over the last 10 years.     

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.     

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 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.