Traditional water mite fixatives and their compatibility with later DNA studies

This work compares frozen water, 70% alcohol, Koenike's and Angelier's fluid as preservation media for water mites in terms of their eventual facilitation of DNA extraction and amplification. The time the mites spent in the fixatives ranged between 1 week and 25 years. Two molecular markers were amplified: 16S ribosomal DNA and CoI mitochondrial DNA. DNA was extractable and could be sequenced from specimens fixed in all the above media, although this generally became more difficult as time progressed. In the light of the known characteristics of these media, the results suggest Angelier's fluid to be the most practical, especially on long expeditions.     

Red, distasteful water mites: did fish make them that way?

Water mites (Acari: Hydrachnida) are unusual among the typically cryptic freshwater fauna in that many species are brightly colored red or orange, and also appear to be distasteful to fish. This apparent aposematism (use of color to warn predators) has been previously explained as the evolutionary end-product of pressure from fish predation. The fish-predation argument has been supported by observations that fish spit out red mites, powder made from red water mites is more distasteful to fish than powder made from non-red mites, and red mites appear to be more abundant than non-red mites in water bodies where fish are present. In this paper, we challenge the hypothesis that fish were the sole driving force behind the evolution of aposematism in water mites. We show that non-red mites actually dominate in water bodies with fish, and that red mites are more abundant in temporary, fishless water bodies. We also demonstrate that powder made from red, terrestrial velvet mites (Trombidiidae) was as distasteful to fish as powder made from red water mites. We suggest that the main role of red and orange carotenoid pigments may be to act as photoprotectants, and hypothesize that redness originated in the terrestrial ancestors of water mites and has been retained in certain lineages of water mites after the invasion of the aquatic habitat. We also suggest that distastefulness evolved subsequent to bright coloration in response to increased conspicuousness to predators. Relaxed selection for redness has occurred when adults and/or larvae are less exposed to sunlight, either through occupying more protected habitats, parasitizing more nocturnal hosts, or parasitizing hosts for a short period of time. Our ability to test this alternative hypothesis is hampered by lack of knowledge of the source and mode of action of distastefulness, and of phylogenetic relationships among the Parasitengona.     

Red, distasteful water mites: did fish make them that way?

Water mites (Acari: Hydrachnida) are unusual among the typically cryptic freshwater fauna in that many species are brightly colored red or orange, and also appear to be distasteful to fish. This apparent aposematism (use of color to warn predators) has been previously explained as the evolutionary end-product of pressure from fish predation. The fish-predation argument has been supported by observations that fish spit out red mites, powder made from red water mites is more distasteful to fish than powder made from non-red mites, and red mites appear to be more abundant than non-red mites in water bodies where fish are present. In this paper, we challenge the hypothesis that fish were the sole driving force behind the evolution of aposematism in water mites. We show that non-red mites actually dominate in water bodies with fish, and that red mites are more abundant in temporary, fishless water bodies. We also demonstrate that powder made from red, terrestrial velvet mites (Trombidiidae) was as distasteful to fish as powder made from red water mites. We suggest that the main role of red and orange carotenoid pigments may be to act as photoprotectants, and hypothesize that redness originated in the terrestrial ancestors of water mites and has been retained in certain lineages of water mites after the invasion of the aquatic habitat. We also suggest that distastefulness evolved subsequent to bright coloration in response to increased conspicuousness to predators. Relaxed selection for redness has occurred when adults and/or larvae are less exposed to sunlight, either through occupying more protected habitats, parasitizing more nocturnal hosts, or parasitizing hosts for a short period of time. Our ability to test this alternative hypothesis is hampered by lack of knowledge of the source and mode of action of distastefulness, and of phylogenetic relationships among the Parasitengona.     

Postharvest disinfestation of diapausing and non-diapausing twospotted spider mite (Tetranychus urticae) on persimmons: hot water immersion and coolstorage

The mortality response of diapausing and non-diapausing twospotted spider mite (Tetranychus urticae Koch) on persimmons to hot water immersion treatments between 44 and 54 °C was examined, for potential as a quarantine treatment. The mean immersion time for mean 99% mortality (LT₉₉) of diapausing mites at 44 °C was 211 min, and this time decreased with increasing temperature to 3.6 min at 54 °C. Non-diapausing mites were found to be less tolerant to temperatures below 48 °C, with an estimated LT₉₉ of 102 min at 44 °C, but had similar thermotolerance above 48 °C. In 47 °C water the immersion time required to kill 99% of diapausing mites was estimated at 67 min. This time was not reduced by subsequent coolstorage at 0 °C for up to eight weeks. Rather, coolstorage had the effect of keeping mites alive, relative to LT₉₉ estimates calculated for mites stored at 20 °C. Similarly the thermotolerance of mites did not change with increased time in diapause, even though mites in diapause for 12 weeks had high control mortality. Hot water immersion appears to be a potentially useful disinfestation method for persimmons.     

DNA标记技术在螨类系统学研究中的应用

长期以来,螨类主要依靠其形态特征进行系统学研究。DNA标记是指能反映生物个体或物种间基因组中某种差异特征的DNA片段。近年来,DNA标记技术在螨类系统学研究中得到越来越广泛的应用。本文综述了随机扩增多态性RAPD、限制性内切酶片段长度多态性RFLP、微卫星SSR、核酸序列扩增、扩增片段长度多态性AFLP和直接扩增片段长度多态性DALP等6种DNA标记技术在螨类系统学研究中的应用现状及前景。     

Oribatid Mites as Potential Vectors for Soil Microfungi: Study of Mite-Associated Fungal Species

The ability of soil-living oribatid mites to disperse fungal propagules on their bodies was investigated. Classical plating methods were applied to cultivate these fungi and to study their morphology. Molecular markers were used for further determination. The nuclear ribosomal large subunit and the nuclear ribosomal internal transcribed spacer of DNA extracts of the cultured fungi as well as total DNA extracts of the mites themselves, also containing fungal DNA, were amplified and sequenced. Based on phylogenetic analysis, a total of 31 fungal species from major fungal groups were found to be associated with oribatid mites, indicating that mites do not selectively disperse specific species or species groups. The detected taxa were mainly saprobiontic, cosmopolitan (e.g., Alternaria tenuissima), but also parasitic fungi (Beauveria bassiana) for whose dispersal oribatid mites might play an important role. In contrast, no mycorrhizal fungi were detected in association with oribatid mites, indicating that their propagules are dispersed in a different way. In addition, fungi that are known to be a preferred food for oribatid mites such as the Dematiacea were not detected in high numbers. Results of this study point to the potential of oribatid mites to disperse fungal taxa in soil and indicate that co-evolutionary patterns between oribatid mites and their associated fungi might be rare or even missing in most cases, since we only detected ubiquitous taxa attached to the mites.     

Random amplified polymorphic DNA analysis of kinship within host-associated populations of the symbiotic water mite Unionicola foili (Acari: Unionicolidae)

Kinship relations within populations of unionicolid water mites are not well known, owing to their complex life cycles and the fact that interactions between active and resting stages for some species are transitory. A number of species of unionicolid water mites are, however, obligate symbionts of freshwater mussels and spend most of their life cycle in association with these hosts. Among these species of mites, parents and offspring are more likely to co-occur and thus provide opportunities to address questions related to the structure of the mating system. The present study employs random amplified polymorphic DNA (RAPD) analysis to address kinship within populations of Unionicola foili living in symbiotic association with the host mussel Utterbackia imbecillis. DNA was amplified from adult mites and a representative number of eggs or larvae (n = 20-30) that were removed from mussels collected on three separate occasions (July, November, and March) over a 12-month period. Parsimony analyses of the molecular data for adults and progeny collected from mussels during July, November, and March revealed distinct groupings, that for the most part, corresponded to mites collected from each of the sampling periods. Many of the genetic markers obtained for male and female U. foili were not evident among the larvae or eggs, suggesting that adults obtained from a host mussel at the time of collection were not the parents of a majority of the progeny. However, female mites and eggs collected from mussels during March and November shared more markers than did females and progeny examined during July. Furthermore, many offspring in the July sampling period were found to have one or more parents absent from the sampled population. Overall, RAPD profiling appears to have limited usage in determining kinship within populations of U. foili, due to its recruitment patterns, and the relatively large number of adults and progeny per mussel. It may, however, prove to be a useful method for assessing genetic relatedness among unionicolid mussel-mites that have substantially lower population densities.     

Complete sequence analysis of 18S rDNA based on genomic DNA extraction from individual Demodex mites (Acari: Demodicidae)

The study for the first time attempted to accomplish 18S ribosomal DNA (rDNA) complete sequence amplification and analysis for three Demodex species (Demodex folliculorum, Demodex brevis and Demodex canis) based on gDNA extraction from individual mites. The mites were treated by DNA Release Additive and Hot Start II DNA Polymerase so as to promote mite disruption and increase PCR specificity. Determination of D. folliculorum gDNA showed that the gDNA yield reached the highest at 1 mite, tending to descend with the increase of mite number. The individual mite gDNA was successfully used for 18S rDNA fragment (about 900 bp) amplification examination. The alignments of 18S rDNA complete sequences of individual mite samples and those of pooled mite samples ( ≥ 1000mites/sample) showed over 97% identities for each species, indicating that the gDNA extracted from a single individual mite was as satisfactory as that from pooled mites for PCR amplification. Further pairwise sequence analyses showed that average divergence, genetic distance, transition/transversion or phylogenetic tree could not effectively identify the three Demodex species, largely due to the differentiation in the D. canis isolates. It can be concluded that the individual Demodex mite gDNA can satisfy the molecular study of Demodex. 18S rDNA complete sequence is suitable for interfamily identification in Cheyletoidea, but whether it is suitable for intrafamily identification cannot be confirmed until the ascertainment of the types of Demodex mites parasitizing in dogs.     

Heat tolerance of strawberry tarsonemid mite Phytonemus pallidus

The heat tolerance of the strawberry tarsonemid mite Phytonemus pallidus ssp. fragariae was studied in northern Sweden by treating adult mites in hot water at temperatures 44°C, 46°C and 48°C. During the summer, the shortest exposure times required to kill all adult mites at these temperatures were 6, 2.5 and 1.25 min, respectively. It is concluded that hot‐water treatment of strawberry runners for 6.5 min at 46°C would suffice to kill all tarsonemid mites on infested runners. This exposure time includes the time needed to raise the temperature of the plant material to that of the water bath. In the autumn, from the middle of September and later, the mites became successively much more heat tolerant as they prepared for hibernation. It is expected that longer exposure periods are needed to kill mites on frigoplants.     

Forensic acarology: an introduction

Mites can be found in all imaginable terrestrial habitats, in freshwater, and in salt water. Mites can be found in our houses and furnishings, on our clothes, and even in the pores of our skin—almost every single person carries mites. Most of the time, we are unaware of them because they are small and easily overlooked, and—most of the time—they do not cause trouble. In fact, they may even proof useful, for instance in forensics. The first arthropod scavengers colonising a dead body will be flies with phoretic mites. The flies will complete their life cycle in and around the corpse, while the mites may feed on the immature stages of the flies. The mites will reproduce much faster than their carriers, offering themselves as valuable timeline markers. There are environments where insects are absent or rare or the environmental conditions impede their access to the corpse. Here, mites that are already present and mites that arrive walking, through air currents or material transfer become important. At the end of the ninetieth century, the work of Jean Pierre Mégnin became the starting point of forensic acarology. Mégnin documented his observations in ‘La Faune des Cadavres’ [The Fauna of Carcasses]. He was the first to list eight distinct waves of arthropods colonising human carcasses. The first wave included flies and mites, the sixth wave was composed of mites exclusively. The scope of forensic acarology goes further than mites as indicators of time of death. Mites are micro-habitat specific and might provide evidential data on movement or relocation of bodies, or locating a suspect at the scene of a crime. Because of their high diversity, wide occurrence, and abundance, mites may be of great value in the analysis of trace evidence.     

A DNA extraction procedure that allows mite specimens to be slide mounted: phytoseiid species evaluated as a model

Four protocols for extracting DNA from mites, using phytoseiid species as exemplars, were evaluated to determine whether the DNA obtained could be used to amplify nuclear, mitochondrial or Random Amplified Polymorphic DNA (RAPD) markers from males, females and eggs. Protocol 3 was identified as the best and this allowed High-fidelity PCR (Hf-PCR) and Hf-RAPD PCR to be used successfully; it left behind the intact body of adult mites so they could be slide mounted for morphological analyses, although the eggs had to be pricked in order to yield sufficient DNA for amplifications. Protocol 3 involved soaking intact specimens in a GuSCN buffer and using a silica matrix, which binds nucleic acids, to yield DNA for amplification. The DNA isolated could be stored up to a month, indicating that the quality was good. This DNA extraction protocol will allow researchers to collect mites, store them in 95% ethanol, and subsequently extract sufficient DNA from single adults or eggs to provide diagnostic PCR products from both nuclear and mitochondrial DNA, yet leave the bodies intact for morphological analyses.     

人体蠕形螨的DNA提取与随机引物PCR检测

【目的】探索人体毛囊蠕形螨和皮脂蠕形螨DNA的提取方法。【方法】采用液氮反复冻融研磨法破碎螨体细胞, 选用改良小昆虫DNA提取法、碱裂解法和试剂盒提取法, 分别提取冻存时间在5个月内和8~10个月的毛囊蠕形螨和皮脂蠕形螨基因组DNA, 并用随机引物PCR方法进行检测。【结果】蛋白核酸测定仪检测结果显示, 试剂盒法提取的DNA纯度较高、量较多, 明显优于改良小昆虫法和碱裂解法。随机引物扩增结果显示清晰的DNA指纹图谱, 两种人体蠕形螨DNA指纹具有明显差异。蠕形螨冻存时间影响DNA提取的量, 但对DNA提取的纯度和RAPD指纹图谱影响较小。不同DNA提取方法提取的同一种蠕形螨DNA指纹图谱基本相似, 试剂盒法和改良小昆虫法提取的DNA样本条带多而清晰, 碱裂解法提取的样本条带少而模糊。【结论】液氮反复冻融研磨法破碎蠕形螨细胞是有效的, 蠕形螨冻存时间不宜超过6个月, 试剂盒提取法是提取蠕形螨DNA的好方法。RAPD技术可以用于这两种人体蠕形螨DNA分子水平上的检测和分类。     

Mite growth on fungus under various environmental conditions and its potential application to biofilters

The effects of relative humidity, temperature, pH and vapor-phase toluene concentration on Tyrophagus putrescentiae growth on Cladophialophora sp. were tested in controlled environmental chambers. It was observed that the mites were able to reproduce readily at relative humidities between 90% and 97% as well as on porous perlite support material pre-soaked in nutrient media of pH 2.5, 4 and 7. Also, the presence of toluene at gas-phase concentrations of 500 to 2000 mg m⁻³ was found to be non-toxic to the mites. The mites, however, were unable to maintain a large population when the temperature was maintained at 14 °C, and overpopulation of the living space led to declines in mite population over time. Overall, it was found to be relatively simple to cultivate mites that may be used for fungal biomass control measures in biofilter applications. This revised version was published online in July 2006 with corrections to the Cover Date.     

一种对甲螨无形态损伤的DNA提取技术

甲螨是一类重要的土壤动物,体型微小,一般具有较厚的体壁。本研究针对甲螨这一特定类群,探讨了一种无形态特征损伤的DNA提取技术。通过结合试剂盒DNA提取法,并适当改进实验条件,设计出一套行之有效的DNA提取流程。通过对提取DNA之后的标本进行形态学观察,发现其主要的分类学特征均保存完好,可以作为凭证标本长期保存。本研究所提供的DNA提取技术既可以提取出足够的DNA又可以保留凭证标本,因此能有效促进甲螨分子分类学相关研究。     

The 'island method'– a new laboratory test method to determine side-effects of plant protection products on the predatory mite Typhlodromus pyri Scheuten (Acari, Phytoseiidae)

Abstract: The model introduced here is an open test which produces experimental results of the same quality as a former test model and at the same time achieves considerable savings in both time and materials. The characteristic feature of the present method is that the mites are kept on a treated glass surface floating on water. The water forms a natural barrier ensuring that the mites cannot escape from the treated area.     

Random amplified polymorphic DNA analysis of kinship within host-associated populations of the symbiotic water mite Unionicola foili (Acari: Unionicolidae)

Kinship relations within populations of unionicolid water mites are not well known, owing to their complex life cycles and the fact that interactions between active and resting stages for some species are transitory. A number of species of unionicolid water mites are, however, obligate symbionts of freshwater mussels and spend most of their life cycle in association with these hosts. Among these species of mites, parents and offspring are more likely to co-occur and thus provide opportunities to address questions related to the structure of the mating system. The present study employs random amplified polymorphic DNA (RAPD) analysis to address kinship within populations of Unionicola foili living in symbiotic association with the host mussel Utterbackia imbecillis. DNA was amplified from adult mites and a representative number of eggs or larvae (n = 20-30) that were removed from mussels collected on three separate occasions (July, November, and March) over a 12-month period. Parsimony analyses of the molecular data for adults and progeny collected from mussels during July, November, and March revealed distinct groupings, that for the most part, corresponded to mites collected from each of the sampling periods. Many of the genetic markers obtained for male and female U. foili were not evident among the larvae or eggs, suggesting that adults obtained from a host mussel at the time of collection were not the parents of a majority of the progeny. However, female mites and eggs collected from mussels during March and November shared more markers than did females and progeny examined during July. Furthermore, many offspring in the July sampling period were found to have one or more parents absent from the sampled population. Overall, RAPD profiling appears to have limited usage in determining kinship within populations of U. foili, due to its recruitment patterns, and the relatively large number of adults and progeny per mussel. It may, however, prove to be a useful method for assessing genetic relatedness among unionicolid mussel-mites that have substantially lower population densities.     

Larval parasitism of spring-dwelling alpine water mites (Hydrachnidia,Acari): a study with particular reference to chironomid hosts

During faunistic investigations on spring habitats in the alpine National Park Berchtesgaden (Bavaria, Germany), water mites were found to be the group with the highest share of species strongly adapted to springs. At four sample sites at two spring complexes, insect emergence was screened for parasitism by larval water mites. A total of at least 36 host species were recorded as being parasitized by 19 water mite species. As in many other habitats, the most important host taxon was shown to be the nematoceran family Chironomidae, both in the number of species and individuals parasitized. Likewise, the number of water mite species attached to chironomids was high. Further host species were found among the Plecoptera, Trichoptera, Coleoptera, Limoniidae and Empididae (Diptera). These taxa were only parasitized by a single water mite species in each case. For 13 mite species, new hosts were recorded for the first time. For another six species, the known host spectrum could be confirmed and/or supplemented. The parasitological data presented (e.g., prevalences, selected attachment sites on the host, larval phenology, intensity of parasitism) provide, in most cases, basic information concerning previously unknown parasite–host associations. At this time, the reason for the strong crenobiosis in water mites cannot be explained by their parasitism.     

Grooming decisions by damselflies, age-specific colonisation by water mites, and the probability of successful parasitism.

We examined whether host damselflies (Ischnura verticalis) in different stages of development were differentially susceptible to parasitism by larval water mites (Arrenurus pseudosuperior). We found that mites were successful in reaching the parasitic phase more often if they colonised hosts closer to emergence. Thus, we predicted that more mites should colonise damselflies closer to emergence and damselflies closer to emergence should spend more time defending against mites. We found that mites colonised damselflies closer to emergence in one of two experiments, but that damselflies in different stages of development did not differ in time spent defending against mites.     

Seasonal changes in tolerance to cold and desiccation in Phauloppia sp. (Acari, Oribatida) from Finse, Norway

In the alpine region at Finse, Norway, Phauloppia spp. (Acari, Oribatida) inhabit lichens on top of boulders. Adult mites are about 0.5 mm in length and have a mean weight of ca. 15 μg. Temperatures in the lichens may drop below -35 degrees C in winter and increase to 55 degrees C in the summer. Large seasonal variations were recorded in supercooling points and body fluid osmolality. Mean January values of SCPs and osmolality were -35.3 degrees C and 3756 mOsm, while July values were -9.4 degrees C and 940 mOsm, respectively. Thermal hysteresis proteins were present in both summer and winter acclimated mites. In mid-winter, some of the mites survived more than 49 days in a water vapor saturated atmosphere at -19 degrees C, and more than 42 days enclosed in ice at the same temperature.The mites showed high tolerance to desiccation. Specimens collected in October survived up to 23 days at 22 degrees C and 5% RH. The tolerance to desiccation was lower in specimens collected during the winter. Some mites survived the loss of up to 90% of their total water content and were reactivated when given access to water. Length measurements of individual Phauloppia sp. showed that both male and female mites are clearly divided in two size groups, suggesting that they belong to two closely related species or different populations.     

Inhibition of Schiff-Reagent Staining in Chromosomes of Ovigerous Tissues of Anoetids; Whole Mites and Sections

Chromosomes of ovigerous tissue of anoetid species are difficult to stain with Feulgen reagents containing HCl and relatively high concentrations of SO₂. The crystals in somatic cells underlying the cuticle, reported to be guanine, can be removed from whole mites or sections when hydrolysed in 1 N HCl at 60 C for 4 min. The hydrolysate of about 1000 anoetid mites, Histiostoma feroniarum, which had been fixed in Carnoy's (6:3:1) fluid for 2 hr, brought to water and then hydrolyzed, inhibits the test-tube reaction between a Schiff reagent and formalin. Paper chromatography of the hydrolysate indicates that it consists of at least 4 components, one of which co-chromatograms with known guanine. The ultraviolet absorption of a mixture of hydrolysate and Schilf reagent indicates that a colorless complex is formed in such a mixture. The refractory response of ovigerous chromatin to Feulgen's reagent may result from the incomplete removal of crystals or hydrolytic products from mites, as well as from the continued hydrolysis by HCl, and from reduced sensitivity caused by excess SO₂, in some preparations of the staining solution.