Orientia tsutsugamushi dynamics in vectors and hosts: ecology and risk factors for foci of scrub typhus transmission in northern Thailand.

BACKGROUND: Scrub typhus is an important neglected vector-borne zoonotic disease across the Asia-Pacific region, with an expanding known distribution. The disease ecology is poorly understood, despite the large global burden of disease. The key determinants of high-risk areas of transmission to humans are unknown. METHODS: Small mammals and chiggers were collected over an 18-month period at three sites of differing ecological profiles with high scrub typhus transmission in Chiang Rai Province, northern Thailand. Field samples were identified and tested for Orientia tsutsugamushi by real-time PCR. The rates and dynamics of infection were recorded, and positive and negative individuals were mapped over time at the scale of single villages. Ecological analyses were performed to describe the species richness, community structure and interactions between infected and uninfected species and habitats. Generalised linear modelling (GLM) was applied to examine these interactions. RESULTS: The site with the highest rates of human infection was associated with the highest number of infected chigger pools (41%), individual chiggers (16%), proportion of the known vector species Leptotrombidium deliense (71%) and chigger index (151). Chigger species diversity was lowest (Shannon diversity index H': 1.77) and rodent density appeared to be high. There were no consistent discrete foci of infection identified at any of the study sites. The small mammals Rattus tanezumi and Bandicota indica and the chiggers L. deliense and Walchia kritochaeta emerged as central nodes in the network analysis. In the GLM, the end of the dry season, and to a lesser extent the end of the wet season, was associated with O. tsutsugamushi-infected small mammals and chiggers. A clear positive association was seen between O. tsutsugamushi-positive chigger pools and the combination of O. tsutsugamushi-positive chigger pools and O. tsutsugamushi-positive small mammals with lowland habitats. CONCLUSIONS: These findings begin to reveal some of the factors that may determine high-risk foci of scrub typhus at a fine local scale. Understanding these factors may allow practical public health interventions to reduce disease risk. Further studies are needed in areas with diverse ecology.

Microbiota in Rosacea.

Rosacea is a complex facial skin condition associated with abnormal inflammation and vascular dysfunction. Next to the known trigger factors, the role of microbiota in the development and aggravation of rosacea continues to raise interest. Demodex folliculorum mites, Helicobacter pylori, Staphylococcus epidermidis, Chlamydia pneumoniae, and the Demodex-associated bacterium, Bacillus oleronius are microbes that have been linked with rosacea. However, the results of studies which assessed their involvement in the disease have been inconsistent and inconclusive. Microbiological research in many different disciplines exploded in recent years as methods to analyze complex microbial communities at the taxonomic and phylogenetic levels became available. Here, we provide an update on the microorganisms implicated in rosacea and review the potential pathogenic role of microbes in the development of rosacea.

Association between the poultry red mite Dermanyssus gallinae and potential avian pathogenic Escherichia coli (APEC).

Avian pathogenic Escherichia coli (APEC) is a group of strains responsible for avian colibacillosis, an impactful disease for poultry farming. The spread of APEC is mainly horizontal, and insects may play a role in their dissemination. However, no data are available about the interaction of APEC with Dermanyssus gallinae, a major arthropodal parasite of poultry. Escherichia coli was detected in the microbiome of the mite, but no specific data have been published till now. Therefore, the presence of the most diffused APEC-associated serogroups has been assessed by PCR in mites collected from 30 flocks of 21 Italian commercial laying hens farms. Escherichia coli was found in 53.3% of the tested groups, corresponding to 66.7% of farms. The most frequent serogroup was O2, but O8, O78, and O109 were also detected. More detailed investigations were carried out in a laying hen farm that was experiencing colibacillosis by APEC O2. The same serogroup was found in both hens and mites, and the maximum likelihood estimation (MLE) of the infection rate (IR) resulted in 24.39 infected mites per thousand, with a pathogen load of 171.47 E. coli O2 cells per mite. The results strongly support the hypothesis of an association between PRM and APEC, confirmed by the contemporary circulation of the same serogroup in both hens and mites, suggesting the potential of D. gallinae as a reservoir or APEC.

Lactobacillus salivarius A3iob Reduces the Incidence of Varroa destructor and Nosema Spp. in Commercial Apiaries Located in the Northwest of Argentina.

Lactobacillus salivarius A3iob was administered to productive colonies belonging to commercial apiaries of small beekeepers (around 30-50 hives each one), from four departments of the province of Jujuy (Argentina): Yala, Tilquiza, El Carmen, and Los Alisos. The incidence of Varroa destructor and Nosema spp., before and after winter, was monitored during 2 years of study (2014-2015). Depending on the geographical location of each apiary and the application time, a monthly dose of the bacteria (105 CFU/mL) reduced the levels of varroasis between 50 and 80%. Interestingly, L. salivarius A3iob cells remitted the percentage of the mites to undetectable values in an apiary treated with flumethrin (at Yala, Yungas region).On the other hand, the spore levels of Nosema spp. in the lactobacilli-treated colonies also depended on the apiary and the year of application, but a significant decrease was mainly observed in the post-winter period. However, at Rivera (El Carmen's department), no significant changes were detected in both parameters.These results obtained after 2 years of work suggest that delivering L. salivarius A3iob cells to the bee colonies can become a new eco-friendly tool to cooperate with the control of these bees' pests.

First record of Aspergillus oryzae as an entomopathogenic fungus against the poultry red mite Dermanyssus gallinae.

The poultry red mite, Dermanyssus gallinae, is a blood-feeding ectoparasite that affects egg-laying hens worldwide. Strategies to control this parasite have focused in the use of entomopathogenic fungi, such as Metarhizium anisopliae. However, only a few studies have evaluated the use of Aspergillus oryzae to control D. gallinae and none of them have employed native strains. In the work presented here, a novel entomopathogenic fungus was isolated from a dead D. gallinae. The results of phylogenetic analysis showed 100% similarity between the isolated strain and those of two species, A. oryzae and Aspergillus flavus, and 99.82% similarity with A. parvisclerotigenus, which were in the same branch of the Flavi section of the genus Aspergillus. This entomopathogenic fungus was a non-aflatoxin B1 producer, as shown by the presence of aflatoxin B1 in the conidial infection suspension. Morphological features of fungus in comparison with A. oryzae and A. flavus indicated that the isolated strain belonged to A. oryzae, and was named Aspergillus sp. Dg-1. The pathogenicity of Aspergillus sp. Dg-1 on D. gallinae at different life stages was then assessed under laboratory conditions. The experiments showed that the isolated strain significantly increased the mortality rate in adult mites, up to 24.83 ± 2.25, compared to the mortality rates in the control group, which were 15.17 ± 2.75 (P ＜ 0.05). However, Aspergillus sp. Dg-1 did not have pathogenic effects on the second nymph stage of D. gallinae. Our findings demonstrate that Aspergillus sp. Dg-1 has pathogenic effects on D. gallinae in their adult stage, presenting biocontrol potential against D. gallinae.

Desarrollo de una PCR para la detección y cuantificación de la parasitación por Demodex folliculorum en biopsias de neoplasias cutáneas del área periocular / Development of a PCR for the detection and quantification of parasitism by Demodex folliculorum infestation in biopsies of skin neoplasms periocular area

Objetivo. Estandarizar la cuantificación relativa por masa de tejido de la parasitación por Demodex folliculorum procedente de biopsias neoplásicas cutáneas de la zona periocular mediante técnicas de amplificación molecular con el objetivo de poder estudiar la posible relación de la aparición de carcinoma basocelular palpebral con la presencia y densidad del ácaro en trabajos posteriores. Material y métodos. Se desarrolló una PCR cuantitativa a tiempo real con sondas TaqMan. La PCR se probó en una serie piloto de 46 muestras reales de biopsias de carcinoma basocelular de tipo nodular. Resultados. La sensibilidad se situó con un límite de detección de entre 1 y 10 copias/μl. El 50% (23/46) de las biopsias fueron positivas a D. folliculorum. La especificidad fue del 100% confirmado mediante secuenciación. Conclusión. La técnica muestra buenos resultados de sensibilidad y especificidad que la pueden hacer útil como herramienta para estudios causa-efecto de D. folliculorum y basalioma (AU)

Objective. To standardize the relative quantification by mass of tissue parasitism by Demodex folliculorum infestation from neoplastic skin biopsies periocular using molecular amplification to study the possible relationship of the appearance of eyelid basal cell carcinoma with the presence and density of the mite in later works. Methods. A quantitative PCR was developed real-time probes TaqMan. PCR was tested in a pilot 46 actual biopsy samples nodular basal cell carcinoma series. Results. The sensitivity was placed with a detection limit of between 1 and 10 copies / μl. 50% (23/46) of the biopsies were positive for D. folliculorum. The specificity was 100% confirmed by sequencing. Conclusion. The technique shows good results for sensitivity and specificity that can make it useful as a tool for studies of cause and effect D. folliculorum and basal cell carcinoma (AU)

Demodex castoris sp. nov. (Acari: Demodecidae) parasitizing Castor fiber (Rodentia), and other parasitic arthropods associated with Castor spp.

A new species of demodecid mite, Demodex castoris sp. nov. (Acari: Prostigmata: Demodecidae), is described based on adult stages from the skin of the nasal region of the Eurasian beaver Castor fiber Linnaeus, 1758, collected in Poland. This is the first detection of a representative demodecid mite in rodents of the suborder Castorimorpha and also represents the first detection of a skin mite in Eurasian beavers. The new species is a small skin mite (average 173 µm in length) characterized by sexual dimorphism related to body proportions. D. castoris sp. nov. was observed in 4 out of 6 beavers examined (66.6%), with a mean intensity of 10.8 and an intensity range of 2-23 ind. host(-1). This paper also contains a checklist of parasitic arthropods known from Castor spp.

Identification and disruption of bacteria associated with sheep scab mites-novel means of control?

Psoroptes ovis mites, which cause psoroptic mange (sheep scab), were investigated to identify potential bacterial targets for endosymbiont control of sheep scab. In addition, transmission of bacteria to the sheep skin was investigated through the characterisation of bacteria present in P. ovis faecal trails and on the fleece environment by internal transcribed spacer (ITS) sequencing. A diverse range of bacteria was identified in addition to a potential endosymbiont candidate, Comamonas sp, which was detected in P. ovis by both ITS PCR and endosymbiont-specific PCR. Disruption of these bacteria within P. ovis, through the use of antibiotics, was explored; with significant reduction in mean mite survival when administered antibiotic diets compared with controls (LR4 = 23.12, P < 0.001). The antibiotic treatments also significantly affected the bacterial density (CFU/mite) within P. ovis, indicating that mite survival may be linked to the bacterial communities that they harbour. Although antibiotics are not suitable for practical application, these results suggest disrupting bacteria associated with P. ovis should be further investigated for novel control.

Synergistic interaction between the fungus Beauveria bassiana and desiccant dusts applied against poultry red mites (Dermanyssus gallinae).

The poultry red mite, Dermanyssus gallinae, is a major pest in egg production, feeding on laying hens. Widely used non-chemical control methods include desiccant dusts, although their persistence under field conditions is often short. Entomopathogenic fungi may also hold potential for mite control, but these fungi often take several days to kill mites. Laboratory experiments were carried out to study the efficacy of 3 types of desiccant dusts, the fungus Beauveria bassiana and combinations of the two control agents against D. gallinae. There was significant synergistic interaction between each of the desiccant dusts and the fungus, with observed levels of mite mortality significantly higher than those expected for an additive effect (up to 38 % higher). Synergistic interaction between desiccant dust and fungus was found also when different application methods were used for the fungus and at different levels of relative humidity. Although increased levels of mortality were reached due to the synergistic interaction, the speed of lethal action was not influenced by combining the two components. The persistence of the control agents applied separately or in combination did not change over a period of 4 weeks. Overall, combinations of desiccant dusts and fungus conidia seem to hold considerable promise for future non-chemical control of poultry red mites.

Field assessment of Orientia tsutsugamushi infection in small mammals and its association with the occurrence of human scrub typhus in Taiwan.

We conducted an extensive study in Taiwan of Orientia tsutsugamushi (OT) infection in small wild mammals. Field trapping was carried out at six districts in eastern and western Taiwan as well as various offshore islands during the period 2006-2010. A total of 1061 specimens representing 11 rodent species were captured. The presence of OT infection was assessed by indirect immunofluorescence assay and polymerase chain reaction assays of 56-kDa type-specific antigen gene. The chigger infestation rate among the animals was 35% (371/1061). Among these, OT was detected in 64% (238/371) of the chiggers from the infested animals and in the spleens from 273 (34.3%) of 797 animals. Excluding animals in the Suncus murinus group, the antibody positive rate of scrub typhus was 69.1% (477 of 690 of serum samples). The prevalence of OT infection in animals from areas with a low incidence of human cases of scrub typhus was significantly lower than that in rodents obtained from regions with a high incidence of human cases of the disease (44.4%±4.0% vs. 71.2%±9.7%, p<0.001). In Taiwan, the prevalence of OT infection in wild rodents is considerably high and appears to correlate positively with the occurrence of scrub typhus in humans.

Bacillus oleronius and Demodex mite infestation in patients with chronic blepharitis.

To better recognize the pathogenicity of ocular Demodex mites, we analysed Bacillus oleronius infection in patients with Demodex-related chronic blepharitis. The studies were conducted on 68 adult patients, in whom ophthalmological and parasitological tests permitted the distinction of a group of 38 patients with a diagnosis of Demodex-related chronic blepharitis (group 1, including a subgroup 1a with moderate blepharitis and a subgroup 1b with severe blepharitis) and a group of 30 healthy individuals (group 2). In every person studied six eyelashes were epilated from each eye and the number of Demodex per eyelash was scored. In parallel, bacterial culture and isolation allowed their phenotypic and molecular identification. The drug sensitivity of the isolates was tested using E-tests. Intensity of Demodex infestation showed no significant differences between subgroups 1a and 1b. From the epilated eyelashes 23 bacterial isolates were obtained, identified as being B. oleronius. All the studied strains were sensitive to ciprofloxacin, doxycycline and gentamicin. The Demodex mite represents an independent aetiopathogenetic factor in blepharitis. In parallel, the parasite may act as a carrier of B. oleronius bacteria, which most probably function as a co-pathogen in the development of severe forms of blepharitis.

The dog mite, Demodex canis: prevalence, fungal co-infection, reactions to light, and hair follicle apoptosis.

Infection rate, reaction to light, and hair follicle apoptosis are examined in the dogmite, Demodex canis Leydig (Prostigmata: Demodicidae), in dogs from the northern area of Taiwan. An analysis of relevant samples revealed 7.2% (73/1013) prevalence of D. canis infection. Infection during the investigation peaked each winter, with an average prevalence of 12.5% (32/255). The infection rates significantly varied in accordance with month, sex, age, and breed (p < 0.05). Most of the lesions were discovered on the backs of the infected animals, where the infection rate was 52.1% (38/73) (P < 0.05). The epidemiologic analysis of infection based on landscape area factor, found that employing a map-overlapping method showed a higher infection rate in the eastern distribution of Taiwan's northern area than other areas. Isolation tests for Microsporum canis Bodin (Onygenales: Arthrodermataceae) and Trichophyton mentagrophyte Robin (Blanchard) on the D. canis infected dogs revealed prevalence rates of 4.4% (2/45) and 2.2% (1/45), respectively. Observations demonstrated that D. canis slowly moved from a light area to a dark area. Skin samples were examined for cellular apoptosis by activated caspase3 immunohistochemical staining. Cells that surrounded the infected hair follicles were activated caspase3-positive, revealing cell apoptosis in infected follicles via the activation of caspase3.

Correlation between ocular Demodex infestation and serum immunoreactivity to Bacillus proteins in patients with Facial rosacea.

PURPOSE: To investigate correlation between ocular Demodex infestation and serum. DESIGN: A prospective study to correlate clinical findings with laboratory data. PARTICIPANTS: We consecutively enrolled 59 patients: 34 men and 25 women with a mean age of 60.4+/-17.6 years (range, 17-93). METHODS: Demodex counting was performed based on lash sampling. Serum immunoreactivity to two 62-kDa and 83-kDa proteins derived from B oleronius was determined by Western blot analysis. Facial rosacea, lid margin, and ocular surface inflammation were documented by photography and graded in a masked fashion. MAIN OUTCOME MEASURES: Statistical significance based on correlative analyses of clinical and laboratory data. RESULTS: These 59 patients were age matched, but not gender matched, regarding serum immunoreactivity, ocular Demodex infestation, or facial rosacea. There was a significant correlation between serum immunoreactivity and facial rosacea (P = 0.009), lid margin inflammation (P = 0.040), and ocular Demodex infestation (P = 0.048), but not inferior bulbar conjunctival inflammation (P = 0.573). The Demodex count was significantly higher in patients with positive facial rosacea (6.6+/-9.0 vs. 1.9+/-2.2; P = 0.014). There was a significant correlation of facial rosacea with lid margin inflammation (P = 0.016), but not with inferior bulbar conjunctival inflammation (P = 0.728). Ocular Demodex infestation was less prevalent in patients with aqueous tear-deficiency dry eye than those without (7/38 vs. 12/21; P = 0.002). CONCLUSIONS: The strong correlation provides a better understanding of comorbidity between Demodex mites and their symbiotic B oleronius in facial rosacea and blepharitis. Treatments directed to both warrant future investigation.

Experimental study on possible transmission of the bacterium Erysipelothrix rhusiopathiae to chickens by the poultry red mite, Dermanyssus gallinae.

The vector potential of the poultry red mite, Dermanyssus gallinae De Geer (Acari: Dermanyssidae), in relation to chicken erysipelas was investigated under experimental conditions. Chickens were inoculated intramuscularly with the bacterium Erysipelothrix rhusiopathiae, and mites were allowed to feed on the inoculated chickens for 5 days. After 20 days of starvation, the mites were allowed to feed on healthy chickens to enable transmission of bacteria. Blood samples were collected from the birds and analysed for the presence of E. rhusiopathiae, and ELISA tests were performed for seropositivity. The internal presence of E. rhusiopathiae in the mites after feeding of inoculated birds was also investigated. It could not be demonstrated that mites take up and transmit E. rhusiopathiae under the experimental conditions described. However, since there are case reports as well as other in vitro studies indicating the potential of D. gallinae to act as a reservoir and potential vector for infections agents, we cannot exclude the possibility that the red poultry mite transmits E. rhusiopathiae between chickens under field conditions.

The role of eriophyoids in fungal pathogen epidemiology, mere association or true interaction?

A considerable number of plant feeding mites representing different families such as Acaridae, Siteroptidae, Tydeidae, and Tarsonemidae interact with plant pathogenic fungi. While species within the Eriophyoidea appear to be the most common phytophagous mites vectoring virus diseases, little is known of their role in fungal pathogen epidemiology. In the present article, we present two studies on eriophyoid-fungal relationships. The first focusing on the association between Aceria mangiferae and the fungal pathogen Fusarium mangiferae in mango is presented as a case study. The second, as the research is still in a preliminary phase, reports on quantitative and descriptive associations between the cereal rust mite Abacarus hystrix and rusts caused by Puccinia spp. Mango bud tissue colonized with F. mangiferae, and wheat and quackgrass leaves colonized with Puccinia spp., supported significantly higher populations of eriophyoid mites. Both mite species were observed bearing the spores of the respective pathogens on their body integument. Aceria mangiferae vectored the pathogen's spores into the bud, the sole port of entry for the fungal pathogen and the frequency and severity of fungal infection increased in the presence of A. mangiferae. While it appears that eriophyoids are playing a role in fungal epidemiology, clearly further research is needed to enhance our understanding of direct and indirect (plant mediated) interactions between plant pathogens and eriophyoid mites in different plant-pathogen systems.

PCR analysis for Wolbachia in human and canine Demodex mites.

In many skin diseases such as Demodex folliculitis, rosacea- or steroid-induced rosacea Demodex mites are present in abundance and are at least partially held responsible for causing these disorders. Although it is known that these diseases respond well to tetracyclines, it is unclear if this is due to the antiinflammatory effects of the antibiotics or to an antibacterial effect on so far unknown bacteria within the Demodex mites. As in filariasis, where the response to doxycycline can be explained by the presence of Wolbachia within the filarial nematodes, this study was performed to see whether Wolbachia also use Demodex mites as their hosts. Human and canine Demodex mite samples were taken by skin scrapings and tested by PCR for the presence of Wolbachia DNA. Wolbachia pipientis DNA was used as positive control. In none of the DNA extracts, Wolbachia were detected showing no evidence for the presence of these bacteria in Demodex mites. The response of Demodex aggravated or Demodex caused diseases to tetracyclines seems not to be due to the presence of Wolbachia in Demodex mites in contrast to the results seen in filariasis.

Genetic differences in internal transcribed spacer 1 between Dermanyssus gallinae from wild birds and domestic chickens.

We investigated the presence of the poultry red mite or the chicken mite, Dermanyssus gallinae De Geer, Acari: Dermanyssidae, in wild bird populations in four different geographical regions of Sweden. The mites identified as D. gallinae were compared genetically with D. gallinae from egg-producing poultry farms in the same regions. The small subunit (SSU) gene, the 5.8S ribosomal RNA (rRNA) gene and the two internal transcribed spacers (ITS) of the rRNA genes were used in the genetic analysis. All D. gallinae mites had identical SSU rRNA, 5.8S rRNA and ITS2 sequences independent of their origin. By contrast, we identified significant differences in the ITS1 sequences. Based on the differences in the ITS1 sequences, the mites could be divided into two genotypes, of wild and domesticated origin, with no variation within the groups. These results imply that wild bird populations are of low importance, if any, as natural reservoirs of D. gallinae in these four geographical regions of Sweden.

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.

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 benzyl benzoate sprays in killing the storage mite Tyrophagus putrescentiae (Acari: Acaridae).

This study tested the effectiveness of benzyl benzoate (BB) in killing Tyrophagus putrescentiae (Schrank) mites when using the method utilized by regular consumers. The efficacy of the BB sprays was determined in laboratory experiments and semi-field experiments with mattress and mattress pad pieces. The mites were counted and their living status determined at different time points microscopically. In the laboratory experiment, the sprays containing either 0.5 %/0.9 % BB with 70 % ethanol or 0.1 % BB with absolute ethanol were highly efficient, resulting in over 90 % mite mortality within 20-30 minutes. In the semi-field experiment, mimicking the home application, the sprays were applied to pieces of a mattress and a mattress pad, and allowed to affect the area for 30 minutes before thorough vacuuming. The recovery of mites was usually less than 10 %. The sprays containing BB were effective in killing the mites in the laboratory, but success was less prominent in the semi-field tests. This method could be used in testing other compounds for their efficacy in killing mites.