Scab Mite Research Articles

Cutaneous hypersensitivity reactions to Psoroptes ovis and Der p 1 in sheep previously infested with P. ovis—the sheep scab mite

We have previously shown that infestation with Psoroptes ovis induces an IgE response and intense tissue eosinophilia, typical of a Type I hypersensitivity response [Parasite Immunol. 22 (2000) 407]. Intradermal tests (IDSTs) suggest that there are also delayed and Arthus-type responses to this parasite. In order to study the nature of ovine cutaneous reactions to P. ovis, naı̈ve controls and experimentally infested sheep ( n=5) were challenged intradermally with mite antigen. Challenge elicited immediate ( P<0.001) and delayed ( P<0.005) wheal reactions in sensitised sheep. At 6 ( P<0.02) and 30 h ( P<0.001) the predominant infiltrating cells were eosinophils. To explore the role of circulating antibodies, naı̈ve sheep ( n=5) were subjected to Prausnitz–Kustner (PK) tests. These elicited immediate ( P<0.02) but not delayed wheal reactions. At 6 h eosinophils ( P<0.001) dominated the infiltrate. These results suggest that P. ovis allergens provoke an IgE-dependent immediate and late phase response and a cell-mediated eosinophil-rich delayed-type hypersensitivity response (ER-DTH).

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.

Identification of an antigen from the sheep scab mite, Psoroptes ovis, homologous with house dust mite group I allergens.

Infestation of sheep with the ectoparasitic mite Psoroptes ovis, results in a severe allergic dermatitis. Currently, little is known about the allergens/antigens that stimulate the allergic response. We have isolated an 836-bp cDNA from a P. ovis cDNA library which displays strong homology to cysteine proteases and, in particular, to the group I house dust mite allergens Der p 1, Der f 1 and Eur m 1. The cDNA was expressed in Escherchia coli, fused to a hexahistidine tag and the recombinant protein (Pso o 1) purified using a nickel-affinity column. The recombinant Pso o 1 was tested for recognition by immunoglobulin (Ig)G and IgE in serum from P. ovis naïve and P. ovis infested sheep. Using Western blots, both classes of antibody to Pso o 1 were detected in postinfestation serum. In enzyme-linked immunosorbent assays, a pronounced IgG-antibody response to Pso o 1 was detected in five of five sheep 3 weeks postinfestation. The IgE-antibody response to whole mite extract was poor in four of five animals. However, a marked IgE response occurred in the fifth animal, and IgE anti Pso o 1 was detected in the serum.

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.

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.

Cloning and sequence analysis of a cDNA encoding Pso o II, a mite group II allergen of the sheep scab mite (Acari: Psoroptidae).

Psoroptes ovis (Hering), the sheep scab mite, is responsible for psoroptic scabies of cattle and sheep. Reverse translation of 30 N-terminal amino acids of the major P. ovis allergen, previously chosen as a candidate immunogen and identified as a 16 kDa protein yielded a degenerate sequence used to design oligodeoxynucleotide polymerase chain reaction (PCR) primers. Use of the PCR primers with a P. ovis cDNA library succeeded in amplification of a 90 bp cDNA gene fragment that was cloned, sequenced, and used to select unique sequencing/PCR primers. Primer walking generated overlapping subclones which yielded the 588 nucleotide consensus sequence of the cDNA encoding the 143 amino acid P. ovis allergen precursor. Nucleotide and translated sequences of the cDNA were compared with sequences in GenBank and found to be homologous to mite group II allergens Lep d II (formerly Lep d I) of Lepidoglyphus destructor Schrank, DerfII of Dermatophagoides farinae Hughes, Der p II of Derrmatophagoides pteronyssinus (Trouessart), Tyr p II of Tyrophagus putrescentiae (Schrank), Eur m II of Euroglyphus maynei (Cooreman) and Gly d II of Glycophagus domesticus (De Geer). The mature P. ovis allergen is composed of 126 amino acids with a calculated molecular mass of 13,468 Da, three disulfide bonds, and pI of 6.06 with one potential o-glycosylation site at Thr116. We designate the P. ovis 16 kDa protein as Pso o II in conformity with nomenclature for mite group II allergens.

Ectoparasites of livestock and companion animals in New Zealand

Extract Principal livestock species in New Zealand, namely sheep, cattle, goats, pigs, horses and deer, are hosts, collectively to at least 45 species of ectoparasites, whereas companion animals such as dogs, cats, rabbits, guinea pigs and ferrets, share about 30 species. Tenquist and Charleston (2001) provide a host/parasite checklist of all species, together with limited information on distribution and aspects of nomenclature. Many of the parasites are not host-specific and none is restricted to New Zealand. There is only one recorded eradication, that of the sheep scab mite, Psoroptes ovis, but the sheep ked, Melophagus ovinus, is very rare.

The growth of sheep scab lesions in relation to sheep breed and time of the year.

Numerically equal groups of between five and ten young Merino and Dorper sheep were artificially infested with sheep scab mites, Psoroptes ovis. Separate groups of sheep were infested during the early winter of two successive years as well as during the early summer of the second year. Lesion growth was measured at two-weekly intervals for a period of six weeks during the first winter and for eight weeks during the second winter and in the summer. The rate of lesion growth and mean lesion size on Merino sheep was significantly greater than that on Dorper sheep at each two-weekly recording during the winter of both years. The rate of growth of the lesions did not differ significantly between the two sheep breeds during summer, although, with the exception of eight weeks post-infestation, the mean lesion size on Merino sheep was significantly larger than that on Dorper sheep at each assessment.

Microfloral diversity of cultured and wild strains of Psoroptes ovis infesting sheep.

PCR amplification of 16S rDNA was used to determine the diversity of bacteria associated with 3 strains of sheep scab mite, Psoroptes ovis. Eight species of bacteria were identified by phylogenetic analysis of the PCR product sequences. Seven of these species are previously unreported in association with sheep scab mites. Five species were matched to Serratia marcesens, Proponibacteium acnes, Phyllobacterium rubiacearum, Pantoea agglomerans, Curacaobacter baltica, whereas the remaining 3 sequences matched unclassified sequences belonging to the gamma proteobacteria, pseudomonads and streptococci. Bacterial diversity of the in vivo cultured strain was very low and did not match the diversity of the 2 wild collected isolates. The diversity of the bacteria in relation to the disease of sheep scab and the possible importance of these bacteria in the diet of the mites are discussed.

Cutaneous and systemic responses during primary and challenge infestations of sheep with the sheep scab mite, Psoroptes ovis.

Lesional area was estimated and lesional mast cells and eosinophils were counted during primary and challenge infestations of sheep with Psoroptes ovis. In addition, circulating basophils and eosinophils and serum P. ovis-specific immunoglobulin IgE antibody were quantified. Expansion of lesional area was significantly less and serum IgE titres significantly greater in challenge than primary infestations. Lesional mast cell hyperplasia and massive eosinophil infiltration accompanied by raised titres of P. ovis antigen-specific IgE antibody were compatible with an IgE-mediated Type-1 allergic reaction, while detection of lesional basophils was suggestive of cutaneous basophil hypersensitivity. The temporal pattern of lesional mastocytosis and eosinophil infiltration and the role of these cells and serum IgE in the immune response is discussed.

Status of Dorper sheep as hosts of ectoparasites

Seventeen ixodid tick species or subspecies have been collected from Dorper sheep in South Africa. The diseases transmitted or the conditions caused by these ticks in sheep are discussed. Although Dorper sheep may be infested with the scab mite, Psoroptes ovis, this does not seem to cause production losses. These sheep may, however, serve as a source of mite infestation for woolled sheep. The majority of Dorper sheep are infested with the larvae of the nasal bot fly, Oestrus ovis, they may also be subject to myiasis caused by the larvae of two calliphorid flies. In certain regions of the country Dorper sheep are prone to serious fly worry by female Simulium chutteri. Although Dorper sheep may be infested by various louse species these do not seem to reach large numbers or cause production losses.

Identification and purification of a 16-kDa allergen from Psoroptes ovis (Acarina: Psoroptidae).

Psoroptes ovis, (Hering), the sheep scab mite, is the causative agent of an allergic dermatitis of sheep and cattle. Recent studies of the host immune response to this ectoparasite have provided information that suggests control may be achieved by immune intervention. A significant effector in protection of the host from clinical lesions is host behavioral grooming. Host grooming is believed to be intensified by a pruritic immediate hypersensitivity response to mite allergens. Knowledge of potential P. ovis allergens is limited. This article reports on the identification and SDS-PAGE continuous elution purification of a 16-kDa polypeptide that elicits immediate type hypersensitivity in calves and has sequence homology with known group II mite allergens, Lep d 2 of Lepidoglyphus destructor (Schrank), and Der f II of Dermatophagoides farinae (Hughes). This P. ovis allergen appears to be a good vaccine candidate for further study and cloning.

Identification of bacterial species associated with the sheep scab mite (Psoroptes ovis) by using amplified genes coding for 16S rRNA.

This was the first molecular study of the bacterial flora of the sheep scab mite (Psoroptes ovis). A sequence analysis of genes coding for 16S rRNA revealed that Serratia marcescens and bacteria closely related to Staphylococcus intermedius or Staphylococcus chromogens and Alloiococcus otitidis were present. These bacteria were associated with skin lesions, dermatitis, and otitis media caused by P. ovis.

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.

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, susceptability 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.

Immunological control of scab mites: digestive enzymes as candidate compounds

Phosphatases, C4 and C8 esterases, leucine and valine aminopeptidases, n-acetyl-β-glucosaminidase, β-glucosidase, β-galactosidase and β-glucuronidase were active in extracts of scab mites ( Psoroptes spp.) raised on sheep or rabbits. Trypsin and chymotrypsin activities were not detected. Haemoglobin was hydrolysed by a detergent-soluble fraction of the mite extracts in a pH-dependent fashion with an optimum of pH 3–5. Acid proteinase activity was greater in mites raised on rabbits than in those raised on sheep. Inhibitors of cysteine, serine and metallo-proteinases failed to inhibit the hydrolysis of H–Pro–Thr–Glu–Phe–Phe(NO 2)Arg–Leu–OH while pepstatin A, a specific inhibitor of aspartic proteinases, totally inhibited its hydrolysis at a concentration of 1 nM.

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.

Antler Size and Winter Mortality of Elk: Effects of Environment, Birth Year, and Parasites

Correlates with antler size of 215 male elk (Cervus elaphus) that died during 5 winters on the National Elk Refuge, Wyoming, were investigated. The elk were 1.5-14.5 (X̄ = 6.5, SE = 0.19) years old. Males with clinical infestations of scab mites (Psoroptes) had antlers with more points, greater beam circumference, and greater mass than males not afflicted with mites. Development of antlers and infestation with scabies were correlated positively with age. Controlling for age, antler size was correlated with March and April temperatures during the year that they grew (nutritional effect) and weather conditions while the males were in utero. Population size, adult sex ratio, and supplemental winter feeding of elk at the National Elk Refuge exhibited no apparent influences on antler size.

Sheep Scab Mite (Acari: Psoroptidae): General Morphology and Immunocytochemistry Using Serum from Infested Animals

The basic morphology of the digestive, reproductive, and nervous Systems of the sheep scab mite, Psoroptes ovis Hering, was defined by light microscopy. An understanding of the stricture of the digestive system of P. ovis is prerequisite to identification of target antigens in specific regions of the digestive tract using immunocytochemistry. Sera from a mite-infested animal and an animal with repeated mite infestations no longer able to maintain a population of P. ovis (acquired resistance) were used to determine antibody specificity to the mite digestive system, and these were compared with the sera from uninfested animals. Sera from naive animals with mite infestations and those animals with repeated infestations had antibodies that were specific against the digestive tract, whereas sera from uninfested control animals did not demonstrate any antibody specificity to mite antigens. Immunocytochemistry demonstrated that polyclonal antibodies to mite gut antigens were present in infested naive animals as well as in multiply infested animals, indicating that gut antigens have a possible role in host resistance.