Do the microorganisms from laboratory culture spent growth medium affect house dust mite fitness and microbiome composition?

The interaction of house dust mites(HDM)and microorganisms is the key factor in the survival of these mites in human-made environments.Spent growth medium(SPGM)provides the rest of the dict,along with dead mite bodies and microorganisms.SPGM represents a source of microorganisms for the recolonization of mite food and the mite digestive tract.An experiment was performed to observe how adding SPGM to the HDM diet affects HDM population growth,the microbiome composition and the microbial respiration in microcosms.We analyzed American house dust mite(Dermatophagoides farinae)and European house dust mite(Dermatophagoides pleronyssinus)originating from control diets and diets treated with an extract of SPGM from 1-and 3-month-old mite cultures.The microbiome was described using 16S and 18S barcode sequencing.The composition of the bacterial and fiungal microbiomes differed between the HDM species,but the SPGM treatment influenced only the bacterial profile of D.farinae.In the D.farinae microbiome of specimens on SPGM-treated dicts compared to those of the control situation,the Lactobacillus profile decreased,while the Candinium,Staphylococ-cus,Acinetobacter,and Sphingomonas profiles increased.The addition of SPGM extract decreased the microbial respiration in the microcosms with and without mites in almost all cascs.Adding SPGM did not influence the population growth of D.farinae,but it had a variable effect on D.pteronyssimus.The results indicated that the HDM are marginally influenced by the microorganisms in their feces.     

Distribution and abundance of house-dust mites (Acarina: Pyroglycphidae) in Rome,Italy

For the first time in Rome, house-dust mite infestation was studied in 90 randomly selected houses. In each house, mite infestation was assessed in three sites: mattress, bedroom and living room. In total, 87.8% of the sampled houses were positive for dust mites. In the houses infested, 11.4% showed densities of >100 mites/g of dust, 15.2% registered densities between 50 and 99, and in the remaining houses (73.4%), the densities were between 1 and 49 mites/g dust. The percentages of infested houses were positively correlated with the relative humidity (RH) values (r=0.89,P=0.02). At the lowest range of RH (between 46 and 50), the infestation was 50% and at the highest range of RH (between 73 and 78) it was 100%. The mattress was significantly the most infested (71.1%) of the tested sites. Only wool and spring mattresses were infested, and they did not show any significant differences in mite concentrations.Dermatophagoides farinae was the most abundant species (53.1%), followed byGlycyphagus domesticus (34.5%),D. pteronyssinus (5.2%), andEuroglyphus maynei (0.2%);D. farinae was also the most frequent species (56.9%). The remaining specimens (7.0%) were predator species commonly found in houses. The prevalence ofD. farinae in Rome is discussed.     

Population dynamics of mites of the family pyroglyphidae and micromycetes in laboratory cultures

Original data on the study of the population dynamics in allergenic house dust mites of the family Pyroglyphidae (Dermatophagoides pteronyssinus (Trouessart 1897) and D. farinae Hughes 1961) during long (37 weeks) joint cultivation with the micromycetes Aspergillus penicillioides Speg. in simple periodical cultures without addition of food (SPC) with different initial population density of mites are given. The micromycete A. penicillioides Speg., dominating in laboratory cultures of pyroglyphids, was cultivated in parallel without mites. It was found that during joint cultivation of A. penicillioides and mites, population dynamics of D. pteronyssinus and D. farinae in SPC depended on the initial population density of mites, which affected the duration of mite developmental stages and the degree of the maximal population density. Cultures of D. farinae developed more rapidly than cultures of D. pteronyssinus, independently of the initial population density (50 or 200 specimens per gram of the substrate). A high degree of the initial population density in both mite species resulted in the shortening of the lag-phase, in more rapid reaching of the maximal population density, and in the higher degree of the maximal population density. Population density of the fungus A. penicillioides did not depend on the presence of both mite species. On the basis of our own data and literary analysis, we assume that A. penicillioides can affect the ability of mites to explore the trophic substrate, the rate of the population development, and the degree of their maximal population density. Mites, in their turn, did not significantly affect the development of A. penicillioides in our experiments.     

IgE reactivity to Acarus siro extract in Korean dust mite allergic patients

Although specific IgE to the storage mite Acarus siro is often detected, there are no detailed studies on IgE reactivity to A. siro in Korea. This study was undertaken to investigate the cross-reactivity to the mite species Dermatophagoides pteronyssinus, Dermatophagoides farinae, Tyrophagus putrescentiae, and A. siro in Korean mite allergic patients. Specific IgE values were determined for the four mite species and a competitive inhibition test was performed for mite extracts using the ImmunoCAP system. The IgE value to D. farinae was the highest among the four mite species tested. There was a strong correlation in the IgE value between house dust mites (D. pteronyssinus and D. farinae) and between storage mites (A. siro and T. putrescentiae). IgE reactivity to A. siro was inhibited by D. farinae and T. putrescentiae extract. Dermatophagoides farinae extract was the strongest inhibitor of IgE binding to A. siro extract, indicating that IgE reactivity to A. siro extract is a cross-reaction caused by sensitization to D. farinae. Strong IgE reactive components were observed in D. farinae and T. putrescentiae extract by SDS-PAGE and IgE immunoblotting. However, no strong IgE-binding component was observed for A. siro. Dermatophagoides farinae is the main source of mite allergens that cause sensitization in Korea. Serum IgE from some of the house dust mite-sensitized patients showed positive responses to storage mite allergens by cross-reaction. Therefore, it is necessary to pay special attention to the diagnosis of mite allergies.     

Interference in foraging behaviour of European and American house dust mites Dermatophagoides pteronyssinus and Dermatophagoides farinae (Acari: Pyroglyphidae) by catmint, Nepeta cataria (Lamiaceae)

The European and American house dust mites, Dermatophagoides pteronyssinus and D. farinae, have a huge impact upon human health worldwide due to being the most important indoor trigger of atopic diseases such as asthma, rhinitis and atopic dermatitis. Preceding studies have shown that the behavioural response of house dust mites towards volatile chemicals from food sources can be assessed using a Y-tube olfactometer assay. In the current study, we used this assay to investigate, for the first time, the ability of the essential oil of the catmint plant, Nepeta cataria (Lamiaceae), known to repel other ectoparasites affecting human and animal health, to interfere with the attraction of D. pteronyssinus and D. farinae towards a standard food source (fish flakes). Two distinct chemotypes (A and B), enriched in the iridoid compounds (4aS,7S,7aR)-nepetalactone and (4aS,7S,7aS)-nepetalactone, and the sesquiterpene (E)-(1R,9S)-caryophyllene, were used. Initial assays with a hexane extract of fish flakes (FF extract) confirmed attraction of mites to this positive control (P < 0.001 and P < 0.05 for D. pteronyssinus and D. farinae respectively), but when presented in combination with either N. cataria chemotype, tested across a range of doses (10, 1, 0.1 and 0.01 μg), decreasing attraction of mites to their food source was observed as the dose augmented. Our study shows that N. cataria, enriched in iridoid nepetalactones and (E)-(1R,9S)-caryophyllene, exhibits potent repellent activity for house dust mites, and has the potential for deployment in control programmes based on interference with normal house dust mite behaviour.     

Structural biology of mite allergens

Mite allergens contribute to a significant proportion of human allergic symptoms, including asthma and rhinitis. The development of therapies to treat and prevent these symptoms depends largely on our understanding of the properties of these allergens. Much effort has been devoted to determining the structure and organization of mite allergens, particularly of the house dust mites, toward understanding their activities and how they elicit immunological responses in humans. Here, we review the structural biology of the major allergens from two species of house dust mites, Dermatophagoides farinae and D. pteronyssinus, as well as allergens from a storage mite, Blomia tropicalis. The knowledge gained from the structural biology of these allergens will enable progress in producing novel, more effective treatments for mite allergies based on specific immunotherapy approaches.     

Molecular identification of house dust mites and storage mites

Mites are known causes of allergic diseases. Currently, identification of mites based on morphology is difficult if only one mite is isolated from a (dust) sample, or when only one gender is found, or when the specimen is not intact especially with the loss of the legs. The purpose of this study was to use polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) of the ITS2 gene, to complement the morphological data for the identification of mites to the species level. For this, six species were cultured: Dermatophagoides pteronyssinus, D. farinae, Blomia tropicalis, Tyrophagus putrescentiae, Aleuroglyphus ovatus and Glycycometus malaysiensis. Genomic DNA of the mites was extracted, quantified, amplified and digested individually with restriction enzymes. Hinf I and Ple I differentiated the restriction patterns of D. pteronyssinus and D. farinae. Bfa I and Alu I enzymes differentiated B. tropicalis and G. malaysiensis. Ple I enzyme was useful for the differentiation between T. putrescentiae and A. ovatus. Bfa I was useful for the differentiation of G. malaysiensis from the rest of the species. In conclusion, different species of mites can be differentiated using PCR–RFLP of ITS2 region. With the established PCR–RFLP method in this study, identification of these mites to the species level is possible even if complete and intact adult specimens of both sexes are not available. As no study to date has reported PCR–RFLP method for the identification of domestic mites, the established method should be validated for the identification of other species of mites that were not included in this study.     

Clinical significance of dust mite allergens

Dust mites are an important cause of allergic diseases worldwide. The traces of Dermatophagoides farinae and Dermatophagoides pteronyssinus can be found all over the world, while Blomia tropicalis are more common allergenic mites in tropical areas. A variety of allergenic components in dust mites have been found, apart from the different positive rates of IgE reactions in dust mite allergic patients, their biological characteristics, effects on the innate immune system, and especially their distribution characteristics in patients are different. Studying the relationship between dust mite allergens and clinical significance will help for diagnosis of patients and formulation of corresponding Allergen Specific Immunotherapy.

     

The fauna and distribution of house dust mites in residential homes of Bandar Abbas District,Southern Iran

This study was designed to determine the occurrence, distribution and abundance of house dust mites (HDM) in residential homes in Bandar Abbas (Hormozgan Province), because of numerous complaints of allergies in this oriental city. The study area was divided in five sampling zones based on population density and geographical distribution. In each sampling zone 10 houses were randomly selected. A total of 50 home dust samples were collected using a portable vacuum cleaner for 2 min from 1 m² of the surface of mattresses, carpets, sofas and furniture in residential houses. After collection, samples were immediately frozen. Mite species were identified and counted using standard methods and keys. Of the sampled houses 88% (44 houses) were contaminated with at least one HDM species. Three species were identified: Dermatophagoides pteronyssinus (63.1%), D. farinae (32.8%) and D. evansi (4.1%) (Pyroglyphidae). Our findings indicate a relationship between HDM density and moisture and temperature of residential places. The high contamination rate of residential houses (88%) and the favourable environmental conditions for these arthropods stress that they should be considered as important allergic causing agents.     

Synanthropic Mites of Kolkata, India: An Ecological Appraisal

Ecological observations were made on house dust mites of Kolkata as they form a major part of synanthropic mite community. Dust samples were collected with regard to the abundance of mite in relation to certain socio-ecological parameters like, habitat preference, location of house, construction pattern of house, types of mattresses used and the frequency of cleaning of mattresses. Among two different habitats examined, bed dust contained significantly higher mite population (p < 0.01) than the corresponding bedroom floor dust. The density of total mites and glycyphagids are significantly higher in rural houses in comparison to those of urban houses. In contrast, rural houses contained least number of pyroglyphids/g of dust. The density of total mites as well as pyroglyphid mites/g of dust are higher in mud house in comparison to concrete house. The density of total mites, pyroglyphids and glycyphagids are higher in cotton mattress in comparison to that of foam mattress. The frequency of cleaning has a significant effect on reducing mite densities i.e., the more the frequency of cleaning the lesser are the mite densities.     

Fumigant toxicity of cassia bark and cassia and cinnamon oil compounds to <Emphasis Type="Italic">Dermatophagoides </Emphasis><Emphasis Type="Italic">farinae </Emphasis>and <Emphasis Type="Italic">Dermatophagoides pteronyssinus</Emphasis> (Acari: Pyroglyphidae)

The toxicity to adults of the American house dust mite, Dermatophagoides farinae, and the European house dust mite, Dermatophagoides pteronyssinus, of cassia bark and cassia and cinnamon oil compounds was examined using residual contact and vapour-phase toxicity bioassays. Results were compared with those of the currently used acaricides: benzyl benzoate and dibutyl phthalate. The acaricidal principles of cassia bark were identified as (E)-cinnamaldehyde and salicylaldehyde. In fabric-circle residual contact bioassays with adult D. farinae, salicylaldehyde (17.3 mg/m²) and (E)-cinnamaldehyde (25.8 mg/m²) were 2.5 and 1.7 times more toxic than benzyl benzoate (43.7 mg/m²), respectively, based on 24-h LD₅₀ values. The acaricidal activity was more pronounced in benzaldehyde, menthol, α-terpineol, and thymol (70.8–234.3 mg/m²) than in dibutyl phthalate (281.0 mg/m²). Against adult D. pteronyssinus, salicylaldehyde (17.3 mg/m²) and (E)-cinnamaldehyde (19.3 mg/m²) were 2.4- and 2.2-fold more active than benzyl benzoate (41.9 mg/m²). The toxicity of benzaldehyde, menthol, α-terpineol, and thymol (75.3–179.2 mg/m²) was higher than that of dibutyl phthalate (285.1 mg/m²). In vapour-phase toxicity tests with adult D. farinae, the test compounds described were much more effective in closed—but not in open—containers, indicating that the effect of these compounds was largely a result of action in the vapour phase.     

House-dust mite sensitivity among rural and urban asthmatics of West Bengal, India: a comparison

House-dust mite allergy is a fairly common problem in West Bengal among individuals sensitive to dust inhalation. House-dust mites belonging to the genusDermatophagoides are abundant in the homes of asthmatic patients residing in urban as well as rural areas of West Bengal. The frequency of positive skin reaction to different dust-related allergens tested was higher (χ²=5.4777, df = 1;P < 0.05) among patients of urban areas compared with that among the patients of rural areas. Urban patients showed more frequent skin reaction towards cockroach allergen, while rural patients are more sensitive to hay-dust allergen and these are very much related to their local environmental conditions. Analysis of radioallergosorbent test (RAST) results against house dust (HD) and mites reveal that 73 and 90% patients of both urban and rural areas responded positively towardsDermatophagoides pteronyssinus (DP) andDermatophagoides farinae (DF) antigens, respectively. The present study indicates no significant difference in house-dust mite sensitivity and mite levels in homes among the rural and urban asthmatics of West Bengal, India as evidenced from the results of analysis of dust samples, allergy skin test and detection of mite-specific IgE antibodies by RAST.     

Indoor mites and forensic acarology

House dust mites (HDM) have been shown to produce allergens causing atopic allergies in human beings, known in medicine as house-dust-mite allergy or atopy. Over the past 25 years (1981–2006) a survey of dust samples from different places was made in the southern part of Poland, to determine the diversity of indoor acarofauna. In total, 1,532 samples were taken from dwellings, hospitals, libraries, research laboratories, drug-stores, and offices and other workplaces. More than 30 mite species were found of which the most abundant and common were HDM, especially Dermatophagoides farinae. Highest mite densities (g^?1 dust) were noted in dwellings. This survey revealed differences in the occurrence and prevalence of various species of domestic mites between and within (dwellings in) geographical areas. This knowledge may be useful in the field of forensic medicine.     

Monoclonal antibodies against recombinant Der f 3 reveal localization of Der f 3 in the gut and faecal pellets of <Emphasis Type="Italic">Dermatophagoides farinae</Emphasis>

Home dust mite derived materials are known to be a major source of problematic inhalant allergens. The aim of this study was to determine the localization of the group 3 allergen, Der f 3, within Dermatophagoides farinae, in order to assess the relative importance of excreted materials and nonexcreted body components as allergen sources. Recombinant Der f 3 (rDer f 3) was expressed in bacteria and purified as an immunogen for production of monoclonal antibodies (mAb) against it. Dermatophagoides farinae mites and their faecal pellets were embedded in paraffin, and serial sections were immunoprobed with mAb clone 3D3 against Der f 3. D. farinae midgut mucosa, gut contents and faecal pellets were strongly immunopositive for Der f 3. Der f 3 immunoreactive products were not detected in any other internal organs of the mite. These results suggest that Der f 3 allergen may be synthesized in and secreted from the digestive tract and excreted from the mite’s body in the faecal pellets.     

Effectiveness of benzyl benzoate in elimination of house dust mites

The effect of elimination treatment with benzyl benzoate was examined in 30 adults with asthma caused by sensitivity to house dust mites. The patients were randomised into a control and an active group, who treated their mattresses with benzyl benzoate (Acarosan^®). The study lasted 12 months and the effect of the treatment was monitored by monthly dust sampling, analyzed for major allergens fromDermatophagoides pteronyssinus andfarinae by the ELISA method and for guanine by the Acarex^® method. The clinical effect was assessed by measuring lung function, daily peak flows, symptoms and medicine consumption as well as skin prick tests, and specific IgE to mite allergens. For both groups, a significant decrease was observed in house dust mite allergens, but there was no significant difference between the groups. No considerable differences were observed in clinical parameters within or between the groups. A good correlation was observed between ELISA and Acarex^®. but the latter showed a major variation in the different classes. In conclusion. treatment of mattresses with benzyl benzoate had no effect in a group of patients with asthma due to house dust mite allergy. Regular vacuum cleaning of the bed may reduce house dust mite allergen exposure.     

The fauna, number and spatial distribution of mites (Acari) in house dust in Moscow

There is a wide and strong allergenic background in Moscow, which is determined by relatively high population density and regular frequency of the allergenic mites in various premises. 34 species have been identified, 5 of them are allergen producers. The mites of the family Pyroglyphidae are the main component of the house dust fauna. Dermatophagoides pteronyssinus and D. farinae distributed all over the world are predominate. A comparative evaluation of the fauna structure, numbers and frequency of the allergenic mites in the flat dust of the allergic patients and healthy people as well as premises is given. Possible importance of the premises in prevalence and circulation of the allergenic mites in the modern town is estimated for the first time.     

The Potential for a Blood Test for Scabies

Background

Scabies afflicts millions of people worldwide, but it is very difficult to diagnose by the usual skin scrape test, and a presumptive diagnosis is often made based on clinical signs such as rash and intense itch. A sensitive and specific blood test to detect scabies would allow a physician to quickly make a correct diagnosis.

Objective

Our objective was to profile the mite-specific antibodies present in the sera of patients with ordinary scabies.

Methods

Sera of 91 patients were screened for Ig, IgD, IgE, IgG and IgM antibodies to S. scabiei, as well as to the house dust mites Dermatophagoides farinae, D. pteronyssinus and Euroglyphus maynei.

Results

45%, 27% and 2.2% of the patients had measurable amounts of mixed Ig, IgG and IgE that recognized scabies mite antigens. However, 73.6% of the scabies patients had serum IgM that recognized scabies proteins, and all except two of them also had IgM that recognized all of the three species of dust mites. No patient had serum antibody exclusively reactive to scabies mite antigens.

Conclusions

Co-sensitization or cross-reactivity between antigens from scabies and house dust mites confounds developing a blood test for scabies.     

Effects of the fungus Aspergillus penicillioides on the house dust mite Dermatophagoides pteronyss'mus: an experimental re-evaluation

Abstract. In this report the widely-held view that house dust mites benefit from fungal contamination of the dietary substratum is re-examined. The performance of Dermatophagoides pteronyssinus (Acari: Pyroglyphidae) is documented over two successive generations in the presence or absence of the xerophilic fungus Aspergillus penicillioides (Hyphomycetales: Moniliaceae). This fungus reduced survival, development rate, adult length and fecundity of D.pteronyssinus. Detrimental effects of A.penicillioides were proportional to the fungal density. Despite the antagonistic effects of A.penicillioides, a requirement for the fungus was indicated by the poor performance of fungus-free mites in the second generation; sustained culture of D.pteronyssinus in the absence of fungi is probably not possible. It is suggested that fungi may alter the particulate nature of the substratum to the detriment of house dust mites, but also provide micronutrients deficient in the diet.     

Population growth and allergen accumulation of <Emphasis Type="Italic">Dermatophagoides pteronyssinus</Emphasis> cultured at 20 and 25°C

The house dust mites, Dermatophagoides pteronyssinus and D. farinae are cultured commercially and in research laboratories and material is harvested from these cultures to make extracts that are used for diagnosis, immunotherapy and research. Temperature and other climatic conditions can influence population growth rates, dynamics of allergen production, and the associated endotoxin, enzyme and protein levels of the mite material harvested from these cultures. Here we determined how temperature affected these parameters. Dermatophagoides pteronyssinus was cultured at 20 and 25°C at 75% relative humidity, and at 2-week intervals the concentrations of mites, Der p 1 and Der p 2 allergens, endotoxin, and selected enzymes were determined. Mite density increased exponentially but growth rate and final population density were greater at 25°C compared to 20°C. The combined allergen (Der p 1 + Der p 2) concentrations accumulated in the cultures at about the same rate at both temperatures. However, individual Der p 1 and Der p 2 accumulation rates varied independently at the two temperatures. Der p 1 accumulated faster at 20°C whereas Der p 2 accumulated faster at 25°C. The amount of Der p 1 in whole cultures was greater than the amount of Der p 2. The concentration of allergen for washed mites harvested from the cultures was much less than for the whole cultures. Our study demonstrated that temperature is an important factor in population growth and the dynamics of allergen production in cultured mites.     

A comparative study of allergenic and potentially allergenic enzymes fromDermatophagoides pteronyssinus,D. farinae andEuroglyphus maynei

The presence of the enzymatically active allergens equivalent toDer p I (cysteine protease),Der p III (serine protease) and amylase in extracts ofDermatophagoides pteronyssinus, D. farinae andEuroglyphus maynei was determined using appropriate enzymatic techniques. Biochemical equivalents of all three allergens were present in each extract studied. Studies also showed that the mite extracts contained a variety of other biochemically active enzymes including trypsin, chymotrypsin, carboxypeptidase A and B, glucoamylase and lysozyme. Marked differences in the relative concentrations of some of these enzymes in different mite extracts were observed, particularly trypsin and carboxypeptidase A. The enzymes were physicochemically similar to equivalent enzymes from vertebrate and invertebrate sources. Chromatofocusing studies of faecal extracts derived fromD. pteronyssinus andD. farinae showed that several isoforms of each enzyme were present. The data indicated that there were more trypsin isoforms, with pI over a wider range, in extracts prepared fromD. pteronyssinus. Proteases and carbohydrases were also found in extracts prepared from faecally enriched material suggesting that they were endoperitrophic and associated with mite digestion. The data suggest that not only are the group I, III and amylase allergens a consistent feature of most pyroglyphid dust mites but also that other proteases and carbohydrases present in mite faeces are allergenic.