Biotin-avidin amplified ELISA for detection of antibodies to Sarcoptes scabiei in chamois (Rupicapra spp.)

Scabies is a major threat to the well being of mountain-dwelling Bovid hosts, Rupicapra rupicapra and Rupicapra pyrenaica. Severe outbreaks are in progress over a significant part of their distribution area and resource managers demand improved methods to monitor, analyse and possibly forecast the spread and effects of scabies at the population level. An amplified capture enzyme-linked immunosorbent assay was developed to detect antibodies to Sarcoptes scabiei in chamois (Rupicapra spp.) serum. The method used the biotin-avidin amplification system and was validated on a panel of 144 serum samples, of which 40 were obtained from scabietic and 104 from healthy unexposed individuals originating from a scabies-free area. The antigen, a whole body extract of the various developmental stages of S. scabiei, was prepared from mites actively leaving the skin lesions of naturally infested red foxes (Vulpes vulpes). The resulting LAB-ELISA was characterised by 93% sensitivity, 97% specificity and a high degree of repeatibility. A single seroreactor was found amongst 32 chamois affected with skin pathologies other than scabies, including infestations by other Acarina (Trombicula spp. and Ixodid ticks). Antibodies to S. scabiei were present in 26 out of 169 sera (15.4%) obtained by clinically healthy chamois within a scabies outbreak area, indicating that asymptomatic infestations by S. scabiei can be revealed by serological methods in the studied Caprinae hosts.     

Efficacy of ivermectin against Sarcoptes scabiei in pigs

The efficacy of orally administered ivermectin against Sarcoptes scabiei was investigated in pigs harbouring experimentally induced infections. Treatment at dosage rates of 300 and 500 microgram per kg body-weight provided 100% control as assessed by mite populations and clinical signs, while at a dose rate of 180 microgram per kg mite populations were substantially reduced but not eliminated.     

Development and Bayesian evaluation of an ELISA to detect specific antibodies to Sarcoptes scabiei var suis in the meat juice of pigs

Samples of ear scrapings, serum and diaphragmatic muscle were collected from 271 fattening pigs at the slaughterhouse. The scrapings were examined for the presence of mites, and tests for specific antibodies to Sarcoptes scabiei var suis in the serum and meat juice were made with an experimental ELISA. The cut-off value for the meat-juice ELISA was estimated at an optical density of 0.5 by receiver operating characteristic curve analysis, on the basis of the cut-off value for the serum ELISA of 0.4. The results of the three tests were used in a Bayesian model to estimate the characteristics of each test. The specificity of the tests of the ear scrapings was considered to be 1 and their sensitivity was estimated by Bayesian analysis to be 0.86, with a 95 per cent confidence interval (CI) of 0.73 to 0.99. The sensitivity of the meat juice ELISA (0.71, 95 per cent CI 0.6 to 0.8) and its specificity (0.77, 95 per cent CI 0.66 to 0.89) were comparable with the sensitivity (0.73, 95 per cent CI 0.6 to 0.8) and specificity (0.81, 95 per cent CI 0.69 to 0.95) of the serum ELISA.     

Responses of immunoglobulin-secreting cells in the skin of pigs during Sarcoptes scabiei infestation

Investigations using immunofluorescence and immunoperoxidase techniques revealed a gradual, but marked, increase in the numbers of immunoglobulin-secreting cells in the skin of pigs during the development of lesions of sarcoptic mange. This was followed by a marked decrease as the lesions regressed. The rise and fall in numbers of these immunoglobulin-secreting cells were observed in both newborn and older animals as they developed signs of cutaneous disease. IgG-secreting cells were the major immunoglobulin-containing cells, followed by IgM- and then IgA-containing cells in the skin of newborn animals. In older pigs, IgM-secreting cells were most prevalent, followed by IgG- and IgA-secreting cells. In multiple infestations, immunoglobulin-secreting cells in the dermis showed very little increase in numbers following the second infestation. Third, fourth and fifth infestations produced little or no increase.     

Experimental Sarcoptes scabiei infestation in pigs: (1) pathogenesis.

Animals were experimentally infested with Sarcoptes scabiei var suis at weekly intervals between birth and five weeks of age. Excoriations were observed on the luminal surface of the ear seven days after the initial infestation. Encrusted lesions developed in the ears of all pigs between the third and eighth weeks but spontaneously regressed and disappeared by the 14th week. A generalised pruritus, accompanied by focal erythematous skin lesions developed in a majority of pigs between seven and 11 weeks of age. The presence of pruritus was associated with an eosinophilia and histological changes in the skin which were consistent with an allergic reaction. The results are discussed in relation to their diagnostic significance and their importance in the control and eradication of the disease.     

Detection of antibodies in sera of weaned pigs after contact infection with Sarcoptes scabiei var. suis and after treatment with an antiparasitic agent by three different indirect ELISAs

Three commercial enzyme-linked immunosorbent assays (ELISAs) were compared for the detection of antibodies to Sarcoptes scabiei var. suis using experimental sera of six 8-week-old pigs after contact infection with Sarcoptes scabiei var. suis. Six non-infected pigs were monitored as a control group. Blood sera were taken once a week from all animals. After successful infection the pigs were treated with an antiparasitic agent (12 weeks post infection (p.i.)) and the antibody titres were monitored until they were negative. The antibody levels of the experimental pigs reached the cut-off level 5 weeks after introduction of an infected animal to the group and were positive by both the Sarcoptes-ELISA 2001 PIG and the Acar-Test P-ELISA. Four weeks after treatment mean results showed optical densities (% OD) below the cut-off level in the Sarcoptes-ELISA 2001 and 8 weeks after treatment in the Acar-Test P-ELISA. In the Chekit Sarcoptest pigs had elevated antibody levels in comparison to control animals, but ODs remained below the given cut-off level at all times. In a second examination with Chekit Sarcoptest (different lot) and at a lower cut-off level, the sera of most of the piglets tested positive. Eight weeks after treatment, four from six pigs still had positive OD values. Therefore this investigation showed a higher sensitivity for the Sarcoptes-ELISA 2001 and the Acar-Test P-ELISA than for the Chekit Sarcoptest. Different test sensitivities must be considered when serologic methods are used for the diagnosis of swine sarcoptic mange, especially for monitoring and controlling eradication programs.     

Efficacy of ivermectin against live mites and eggs of Sarcoptes scabiei in pigs

Sows infested with Sarcoptes scabiei var. suis (ssvs) were treated with 75, 150 and 300 micrograms/kg of ivermectin by a single subcutaneous injection at the neck region. Compared to the numbers of mites and eggs just before injection, those on post treatment weeks (PTW) 1, 2 and 4 showed significant decreases. Especially at 300 micrograms/kg, the counts showed almost all mites and eggs were eradicated on PTW 1, manifesting ivermectin to possess potential effect on ssve without apparent abnormal side effect. Potential mitocide effect of ivermectin on ssvs was revealed.     

疥螨与疥螨病研究进展

疥螨病通常是指由疥螨科、疥螨属的疥螨(Sarcoptes scabiei)寄生于人和其他哺乳动物皮肤表皮内，引起称为疥疮(又称疥癣、癞病)的一种顽固、接触性、传染性皮肤病，其以剧痒、结痂、脱毛和皮肤增厚为特征。1698年意大利的两位学者首次对疥螨病进行了说明和描述；然而，直到200年后疥螨     

Experimental quantification of the transmission of Sarcoptes scabiei var. suis among finishing pigs

In this study, the rate of S. scabiei var. suis transmission among finishing pigs was quantified in a contact transmission experiment. Forty piglets originating from a mange free farrow-to-finish herd were randomly allocated to three groups and one S. scabiei var. suis infested finishing pig was subsequently added to each of these groups. After 35 days, the three seeder pigs were removed from the groups and the remaining 40 pigs were re-allocated to five pens. Ear scrapings, to be examined for mites, were collected from each pig on days 1, 14, 28, 42, 56, and 84 of the experiment. Blood samples, to be tested for antibodies against S. scabiei, were collected from each pig on days 0, 14, 28, 42, 56, 70, 84 and 112 after the introduction of the seeder pigs.From the results of the ear scrapings and the blood samples the number of susceptible (not infested) and infested pigs was derived at the time of each sample collection and the number of new infestations in the intervals between the sample collections. From these data the infestation rate parameter beta (average number of new infestations per infested pig per day) was estimated by use of a Generalised Linear Model (GLM) and accordingly, beta was estimated at 0.056 (95% CI: 0.037-0.085) infestations per infested pig per day.Next, by use of beta, the transmission of S. scabiei was simulated in a population of 100 finishing pigs for 100 days after the introduction of a single infested pig. For this purpose, 500 simulations were done. The 90% confidence interval of the number of infested pigs at day 100 ranged from 12 to 88 (median: 63). It was concluded that transmission of S. scabiei among finishing pigs is slow. Due to the presumed lower contact rate between sows as compared to finishing pigs, it is anticipated that transmission of S. scabiei among sows will even be slower than among finishers These findings are of particular interest for the development of surveillance programmes for S. scabiei free herds.     

Sarcoptes scabiei infestation in a cat

Sarcoptes scabiei infestation was diagnosed in a cat. Clinical signs included thick, crusty, exudative dermatitis on the feet, caudal aspect of the thighs, and tail. Paronychia and dystrophic nails also were observed. The course of the disease was chronic. Concomitant and potentially predisposing additional health problems were suspected, but were not confirmed.     

Isotype-specific ELISAs for the detection of antibodies to bovine respiratory syncytial virus

Isotype-specific ELISAs for the detection of antibodies to bovine respiratory syncytial virus (BRSV) are described. BRSV-specific IgG1 and IgG2 were determined in indirect double antibody sandwich assays. For IgA and IgM antibody capture assays were used. The isotype specificity of the assays was confirmed by the observation that samples with a high titre of BRSV-specific antibodies of particular isotype were negative in the assays for the other isotypes and vice versa. Comparison of the results obtained in the ELISAs and in the virus neutralisation test showed that acute phase antibodies were more efficiently detected in the latter. It also showed that the presence of BRSV-specific IgA was not correlated with neutralising activity in vitro. The serum antibody response of BRSV-infected seronegative calves from the field consisted of a nearly simultaneous increase of IgM, IgA and IgG1-antibodies in the acute phase of the disease, while the IgG2-response followed at various intervals thereafter. In young animals with maternal antibodies a different pattern was found. There was no increase in IgG1 and IgG2, but six of eight animals showed a weak IgM response and two of these six calves also showed a weak and short lasting IgA response. Because maternal antibodies are insufficiently effective in protecting calves against BRSV, the presence of such antibodies at mucosal surfaces was investigated. Maternal immunity was found to be restricted to IgG1 antibodies in serum. This agrees with the failure of maternal antibodies to protect mucosal surfaces against BRSV infection.     

Sarcoptes scabiei infestation of a donkey in the UK

Sarcoptes scabiei infestation was identified as the cause of pruritic dermatitis in a donkey in the UK. Treatment with i.m. doramectin and topical selenium sulphide was successful in eliminating clinical signs. Foxes were identified as the possible source of infestation. Sarcoptic mange should be considered as a potential differential diagnosis for pruritic dermatitis in equids in the UK.     

Immunoblot analysis of IgG antibody response to Sarcoptes scabiei in swine

This study was performed to determine the frequencies and specificities of IgG antibodies binding to component of Sarcoptes scabiei extracts in swine with hypersensitive and chronic mange. The hypersensitive form is characterised by pruritus and the presence of small red papules over the flanks and belly. The chronic form is characterised by crusts, which contain large numbers of mites and are attached to the skin; the lesions are most commonly found on the internal pinna extending into the auditory canal. S. scabiei mite extract was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with subsequent immunoblotting. IgG-binding proteins were detected with individual sera from 30 hypersensitive and 21 chronically infected pigs; eight "Specific Pathogen Free" pigs were used as negative controls. Seven protein bands with molecular weights ranging from >220 to 30 Kilodalton (KDD) (>220, 218, 110, 80, 66, 52, 36 KDD) strongly bound with IgG antibodies; five out of these seven components (218, 110, 80, 66, 52 KDD) bound also with sera from negative pigs. There is a statistically significant difference in the antigenic recognition spectra between hypersensitive and chronically infected pigs; component of >220 KDD is more frequently recognized by chronically infected pigs (P=0.0006, chi(2)=11.74), in contrast component of 36 KDD is more frequently recognized by hypersensitive pigs (P=0.001, chi(2)=10). Our results clearly indicate there is a difference in the reactivity to antigenic peptides/proteins of S. scabiei mite between hypersensitive and chronically infected pigs, and revealed that only two antigens may be considered S. scabiei-specific and used for diagnostic purposes in swine.     

Transmission of Sarcoptes scabiei in Swine by Fomites

Transmission of sarcoptic mange by fomites was investigated by placing mange-free piglets in pens for either fixed or variable periods of time during the first six days following removal of mange infected swine. Transmission occurred in pigs with as little as 24 hours exposure to fomites. Clinical signs of pruritis and focal erythematous skin lesions developed in various pigs from four and a half to 13 weeks after exposure. Pigs with the longer exposure developed clinical signs more rapidly than those with a shorter exposure. Four of six pigs developed a chronic form of the disease characterized by thickened encrustations and scurf from which mites were readily demonstrated. The remaining two pigs developed only the pruritic form and mites were never found in numerous skin scrapings examined.     

A comparison of two ELISAs for the detection of antibodies to bovine leucosis virus in bulk-milk

OBJECTIVE: To estimate the sensitivity, specificity and detection limits for two bulk-milk enzyme-linked immunosorbent assays, the Svanovir BLV-gp51-Ab and the Lactelisa BLV Ab Bi indirect tank 250, for the detection of antibody to bovine leucosis virus in milk. PROCEDURE: Milk samples from 27 cows known to have enzootic bovine leucosis (EBL) were serially diluted with milk from a herd known to be free from the disease. The dilution at which antibodies could no longer be detected by each test was determined. A total of 1959 bulk-milk samples submitted to a laboratory for the Victorian (EBL) eradication program were tested with both the Svanovir and the Lactelisa assays. A Bayesian approach was used to calculate maximum-likelihood estimates of test sensitivity and specificity. An additional 660 bulk-milk samples were tested with both the Svanovir and the Lactelisa assays. Herds that had positive results on either or both of the assays were subjected to blood or milk testing of individual cattle. RESULTS: The dilution of milk at which the Svanovir assay failed to detect enzootic bovine leucosis antibody in half of the samples was 1 in 40, whereas the comparable value for the Lactelisa was 1 in 200. Computer modeling of the operating characteristics of the Svanovir assay indicated that the sensitivity of that assay would be considerably lower than that for the Lactelisa, and the specificity was estimated to be higher. Evaluation of the assays using 660 bulk-milk samples showed that the Lactelisa assay detected four infected herds that were not detected by the Svanovir test. No false positive results were recorded for either assay. CONCLUSION: Use of the Lactelisa assay in the Victorian EBL eradication program will enhance disease detection and eradication, but may also result in an increased frequency of false positive bulk-milk test results.