Stylostome organization in feeding Leptotrombidium larvae (Acariformes: Trombiculidae)

The stylostome of larvae of the trombiculids Leptotrombidium scutellare (Nagayo et al.), Leptotrombidium fletcheri (Womersley et Heaslip) and Leptotrombidium deliense (Walch) was studied experimentally at different time intervals after larval attachment using the histological method. The stylostome of these species has the same organization and belongs to the epidermal combined with the mixed type, developing more in width than in length. Neither transverse nor conspicuous longitudinal layers are present within the stylostome walls, which stain predominantly in red with Azan, also showing longitudinal portions with blue staining. Larvae tend to attach closely to each other and scabs, consisting of the hyperkeratotic epidermal layers fusing with migrating inflammatory cells, develop around the attachment sites. The dermis shows inflammatory foci with dilated capillaries and inflammatory cells inserting in the connective tissue layer underneath the stylostome. The feeding cavity, which is moderately expressed, may be found either in the epidermis or in the dermis. It contains inflammatory cells and their debris in the liquefied host tissues. The stylostome length depends on the character of the attachment site (the thicker epidermis or scab the longer the stylostome), and does not directly correspond to the stages of larval feeding. Nevertheless, at the 48-h time interval, nearly all attached larvae are found to be fully fed and their midgut cells are filled with nutritional globules.     

无前恙螨属--新种记述(蜱螨目:恙螨科)

本描述了采自江苏马铁菊头蝠(Rhinolophus ferrum-eguinum)翅上的无前恙螨属一新种,即宜兴无前恙螨[Walchia(Walchia)neeshini Wang et Xia,sp．nov]。模式标本保存于复旦大学上海医学院寄生虫学教研室。     

Ultrastructural organization of hypodermis and formation of cuticle in pharate larva of Leptotrombidium orientale (Acariformes: Trombiculidae)

The ultrastructural organization of hypodermis and the process of cuticle deposition is described for the pharate larvae of a trombiculid mite, Leptotrombidium orientale, being under the egg-shell and prelarval covering. The thin single-layered hypodermis consists of flattened epithelial cells containing oval or stretched nuclei and smooth basal plasma membrane. The apical membrane forms short scarce microvilli participating in the cuticle deposition. First of all, upper layers of the epicuticle, such as cuticulin lamella, wax and cement layers, are formed above the microvilli with plasma membrane plaques. Cuticulin layer is seen smooth at the early steps of this process. Very soon, however, epicuticle starts to be curved and forms particular high and tightly packed ridges, whereas the surface of hypodermal cells remains flat. Then a thick layer of the protein epicuticle is deposited due to secretory activity of hypodermal cells. Nearly simultaneously the thick lamellar procuticle starts to form through the deposition of their microfibrils at the tips of microvilli of the apical plasma membrane. Procuticle, as such, remains flat, is situated beneath the epicuticular ridges and contains curved pore canals. Cup-like pores in the epicuticle provide augmentation of the protein epicuticle mass due to secretion of particular substances by cells and to their transportation through the pore canals towards these epicuticular pores. The very beginning of the larval cuticle formation apparently indicates the starting point of the larval stage in ontogenesis, even though it remains for some time enveloped by the prelarval covering or sometimes by the egg-shell. When all the processes of formation are over, hungry larvae with a fully formed cuticle are actively hatched from two splitted halves of prelarval covering.     

Ultrastructure of the integument during moulting of the quiescent tritonymphal instar of trombiculid mite Hirsutiella zachvatkini (Acariformes: Trombiculidae)

The ultrastructure of the integument of the quiescent reduced tritonymph of the trombiculid mite Hirsutiella zachvatkini (Schluger) was investigated by means of transmission electron microscopy. Mites were investigated daily during the 14–16 day tritonymphal period (imagochrysalis). This period includes the deutonymphal moult (1–3 days), the quiescent tritonymph period (2–4 days), and the tritonymphal moult into the adult mite (6–10 days). A distinct recognizable feature of the tritonymphal moulting cycle is a sequence of events independent of precise time intervals. This process involves partial destruction and reorganization of the hypodermis of the previous instar, and formation of a new hypodermis of the subsequent instar from islands of rudimentary hypodermal cells. The integument of the reduced tritonymph differs greatly from that of both larva and active deutonymph and adult. It consists of a simply organized hypodermal layer of varying thickness and a thick clear poorly lamellate cuticle with curved pore canals, and lacking setae. The epicuticle is very thin and without a clear protein layer. The tritonymphal instar as such with its own cuticle situated near the hypodermis is encased within the detached covering of the previous active deutonymph, and may be considered a calyptostasic and entirely pharate instar. There is a tendency for reduced tritonymphal stage to be eliminated from ontogenesis and this stage is not homologous to the pupa of insects.     

Stylostome formation in trombiculid mites (Acariformes: Trombiculidae)

Stylostomes of the trombiculid mite larvae Neotrombicula pomeranzevi (Schluger), Hirsutiella zachvatkini (Schluger), Miyatrombicula esoensis (Sasa and Ogata) and Euschoengastia rotundata (Schluger) (Acariformes: Trombiculidae), formed in the host skin during feeding of the parasites on their natural hosts (voles) were studied histologically and histochemically. A stylostome is a variously shaped tube formed of solidified mite saliva that extends from the mouthparts of the parasite through the epidermis into the dermis of the host, and allows the mite to obtain its liquid food. The first step of stylostome formation is deposition of an eosinophilic cone, to which the larva’s chelicerae are glued. Organization of the stylostome depends on the mite species, and its walls may show weakly expressed longitudinal or transverse stratification. Histochemically, the stylostome is composed of complex glycoprotein with varying tinctorial properties through the width or the length of the stylostome’s walls. Beneath the distal end of the stylostome, irrespectively of its localization either in the epidermis or in the dermis of the host, a feeding cavity is formed as a result of the action of the hydrolytic components of the mite’s saliva forced through the stylostome into the wound. An inflammatory dermal reaction of moderate intensity is evolved during larval feeding and stylostome formation. It is manifested by the infiltration of the foci with neutrophiles, lymphocytes and macrophages and by dilation of capillaries of the terminal vessel bed and filling them by erythrocytes and other blood elements. Around the stylostome, necrosis of the epidermal cells occurs, leucocytes come to the damaged area and fuse with the necrotic epidermal cells, leading to the formation of the large scabs on the surface of the host’s skin. In the case of E. rotundata, single capsules having a terminal opening and containing feeding larva are formed on the abdomen of the hosts. The walls of the capsules are composed of the mite’s saliva flowing upon the surface of the host’s skin. At the bottom of the capsule, a stylostome perforating the epidermis is also present.     

Functional morphology and ultrastructure of mouthparts in unfed water mite larvae Piona carnea (Koch, 1836) (Acariformes: Pionidae)

Mouthparts of unfed larvae Piona carnea (Koch, 1836) (Acariformes: Pionidae) were studied on whole-mount preparations, semi-thin sections and with TEM and SEM methods. The mouth apparatus is incorporated within the pseudotagma, gnathosoma, composed of the infracapitulum and of the chelicerae resting on the roof of the latter. The gnathosoma inclines to the long axis of the body and is inserted at its base into the idiosoma by the circumcapitular fold. The basal cheliceral segments are long and fused. An anterior projection, the proposed fused fixed digits, protrudes from the distal end of the basal cheliceral segment forward between the movable digits. The movable digits are always found in protruded position, strongly curved upward and show a groove on their inner sides. The ventral wall of the infracapitulum is made of the mentum posterior and the malapophyses anterior to the palp articulation. The malapophyses are squeezed between the large palps and envelope the distal portion of the chelicerae from the sides. The ventral portion of the fused malapophyses are provided with a characteristic ventral cuticular fork of unknown function. Each malapophysis terminates by a flexible lateral lip provided by several rigid jags looking posterad. The palps face downward and backward, and bear on the tibia the large curved palpal claws turned laterad. The palp femur bears on the ventral aspect a characteristic wide spade-like projection provided with its own muscles originating on the dorsal wall of the femur. The labrum is a thick cuticular arrow-like structure protruding forward into the preoral cavity, whereas the cervix is a thin weakly sclerotized plate. The particular labral valve projects forward from the dorsal basis of the labrum into the preoral cavity. The labrum and the cervix are provided by their own small labral and cervical muscles originating on the cervical apodemes. The pharynx is totally separated from the ventral wall of the infracapitulum and is devoid of ventral dilators. The dorsal pharyngeal dilators originate on the thick and sclerotized capitular apodeme and, posteriorly, on the paired cuticular branches, capitular apodemes, which end freely in the body cavity and are combined with the common salivary duct. The short sigmoid pieces serve for origin of the levator muscles of the chelicerae. Retraction of the gnathosoma and the chelicerae is mediated by several sets of muscles originating on the dorsal plate.     

Comparative ultrastructure of coxal glands in unfed larvae of Leptotrombidium orientale (Schluger, 1948) (Trombiculidae) and Hydryphantes ruber (de Geer, 1778) (Hydryphantidae)

Coxal glands of unfed larvae Leptotrombidium orientale (Schluger, 1948) (Trombiculidae), a terrestrial mite parasitizing vertebrates, and Hydryphantes ruber (de Geer, 1778) (Hydryphantidae), a water mite parasitizing insects were studied using transmission electron microscopy. In both species, the coxal glands are represented by a paired tubular organ extending on the sides of the brain from the mouthparts to the frontal midgut wall and are formed of the cells arranged around the central lumen. As in other Parasitengona, the coxal glands are devoid of a proximal sacculus. The excretory duct, joining with ducts of the prosomal salivary glands constitutes the common podocephalic duct, opening into the subcheliceral space. The coxal glands of L. orientale are composed of a distal tubule with a basal labyrinth, an intermediate segment without labyrinth, and a proximal tubule bearing tight microvilli on the apical cell surface and coiled around the intermediate segment. The coxal glands of H. ruber mainly consist of the uniformly organized proximal tubule with apical microvilli of the cells lacking the basal labyrinth. This tubule shows several loops running backward and forward in a vertical plane on the side of the brain. In contrast to L. orientale , larvae of H. ruber reveal a terminal cuticular sac/bladder for accumulation of secreted fluids. Organization of the coxal glands depends on the ecological conditions of mites. Larvae of terrestrial L. orientale possess distal tubule functioning in re‐absorption of ions and water. Conversely, water mite larvae H. ruber need to evacuate of the water excess, so the filtrating proximal tubule is prominent.     

Electron microscopic observations of Orientia tsutsugamushi in salivary gland cells of naturally Infected Leptotrombidium pallidum larvae during feeding

We performed a detailed electron microscopic observation on the escaping process of Orientia tsutsugamushi from the salivary gland cells of naturally infected trombiculid larvae into the acinar lumen of the gland during feeding on mice. In unfed larvae, many O. tsutsugamushi were intermingled with secretory granules in the cytoplasm of the salivary gland cell. O. tsutsugamushi was neither found in the acinar lumen nor observed escaping from the apical surface of the gland cell. In contrast, in the larvae fed on mice, many O. tsutsugamushi were observable in the acinar lumen. They were enveloped with the host glandular cell membrane. In salivary gland cells, secretory granules changed the distribution and accumulated in the apical region. In such cells, the majority of O. tsutsugamushi were found at the base of the cell. Some O. tsutsugamushi were pushing the glandular cell membrane outward in various degrees, showing different stages of escape. These findings suggest that larval feeding induced O. tsutsugamushi escape from salivary gland cells, that the escape was by budding, during which O. tsutsugamushi were enveloped in the host cell membrane, and that O. tsutsugamushi would be injected into the mouse skin as a mixture with mite saliva. The study also revealed the presence of many small vesicles that had the same cell wall structure as O. tsutsugamushi in the cytoplasm of the salivary gland cell. Most of them seemed to be products from degenerated Orientia.     

Transovarial Transmission of Orientia tsutsugamushi in Leptotrombidium palpale (Acari: Trombiculidae)

Transovarial transmission of Orientia tsutsugamushi in colonies of Leptotrombidium palpale was studied in the parent and F₁ and F2 generations. Both transovarial transmission and filial infection rates were 100% in the parent and F₁ generations of Leptotrombidium palpale. The filial infection rate in the F₁ generation was 100%, but it declined to 94.3% in the F₂ progeny. The sex ratio of the F₁ generation from infected L. palpale was 1∶0.8 (male:female) and the proportion of males was relatively high. This study is the first to report on the transovarial transmission of O. tsutsugamushi in L. palpale. High transovarial transmission rates in L. palpale suggest that this species might be one of the major vectors of tsutsugamushi disease in Korea.     

Stylostome formation by Leptotrombidium mites (Acari: Trombiculidae).

The modes of stylostome formation by larvae (chiggers) of Leptotrombidium intermedium, L. fletcheri, L. arenicola, and L. deliense in parasitized mouse skin were studied histologically in relation to their capacity to transmit Rickettsia tsutsugamushi. Three types of stylostome formation were recognized among the different species: the epidermal stylostome formed by the larva of L. intermedium; the mesenchymal stylostome formed by the larva of L. fletcheri; and, the mixed stylostome formed by the larva of both L. arenicola and L. deliense. Dermal inflammations related to the three types of stylostomes were histologically defined. The possible importance of stylostome characteristics to the transmission of rickettsial organisms is discussed.     

The origin of parasitism in trombiculid mites (Acariformes: Trombiculidae)]

On the basis of literary data and original investigations some phylogenetic, ecological and morphological aspects of the origin of parasitism in trombiculid mites are carefully considered for the first time. It is shown that parasitism in this group of trombidiform mites is a relatively young historical phenomenon and was formed after their ontogenesis had differentiated into active and quiescent stages. Therefore, in the life pattern of trombiculid mites the character of individual development, that defines their biotopical restriction, is much more important than the phase parasitism. Primitive organization of the digestive system and extraintestinal digestion, so characteristic of this group, are one of the main reasons of the origin of their parasitism. Under pasture conditions trombiculid mites, that initially were predators-entomophages with bite-sucking mouth parts, pass easily to parasitism on vertebrate animals and become primary lymphophages. They use the vertebrate host's organism exclusively as a source of food and by the extent of polyphagia are very close to free-living blood-sucking insects. Stylostome, that develops during feeding of trombiculid larvae and some other closely related groups of trombidiform mites, is a universal structure for achieving a large amount of food on a wide range of animals during a relatively short period of time and reflects wide host-parasite specificity of these parasitic mites. From the historical view the larvae of trombiculid mites did not pass from one group of hosts to the others, but owing to morphological preadaptation to parasitism passed in a definite historical period, not earlier than Paleogene, to parasitism on all classes of terrestrial vertebrates, especially on mammals, their primary hosts.     

Characterization of the initial ontogeny of Leiarius marmoratus (GILL, 1870): larvae to juvenile

Aiming to provide data on the biology of Leiarius marmoratus, which will aid in its production in captivity, as well as in studies for its preservation in the environment, this work had as objectives: analyze and describe main morphological alterations during larval ontogeny of the species. We analyzed 205 individuals, obtained by induced reproduction (Colpani Pisciculture) and kept in CEPTA/ICMBIO, Pirassununga, São Paulo, Brazil. Analyses were performed from hatching moment to 30th day. The specimens were classified into two periods: larval (Stages: vitelline, pre‐flexion, flexion, post‐flexion) and juvenile. Hatched larvae showed ident chromatophores only at anterior and posterior extremities of yolk sac. The standard length ranged from 2.16 mm (yolk) to 28.84 mm (Youth). Dorsal fin rays were initially observed at flexion stage (12–14 rays). Major alterations occurred during post‐flexion/juvenile stage, when dorsal, pectoral, pelvic, anal, and caudal fins were observed and pigmentation intensified throughout the lateral region, forming bands in the body, one between the end of the head and beginning of dorsal to pelvic fin, and another one beginning at dorsal to caudal peduncle and four longitudinal at the head.     

Morphological variablility in the chigger mite species Neotrombicula sympatrica (Acariformes: Trombiculidae) from Kyrgyzstan

Aberrations (quantitative chaetotactic deviations, i.e. decreasing or increasing of setae numbers and variations in arrangement of setae) and anomalies (qualitative chaetotactic deviations, for example, partial reduction of scutum, shortening of a seta more than 1.5-2 times, merging of setae) were recorded for 13 taxonomically important morphological structures in the chigger mite species Neotrombicula sympatrica Stekolnikov, 2001. 3308 specimens were studied as a total. 17.2% of them had various morphological deviations. The most common types of aberrations were observed in the number and positions of genualae I (94 specimens), AM seta (79 spec.) and sternal setae (77 spec.). The aberrations of sternal and coxal setae were usually interrelated: the sternal seta was "transferred" from the sternal area onto the coxa, or the other way round take place. The specimens having aberrations of sternal setae were twice as numerous as the specimens with aberrations of coxal setae (77 against 35). The specimens with aberrations of dorsal setae and mastitarsala were very rare (2 spec. each). Among anomalies, the presence of nude galeal seta (91 spec.) and scutal anomalies (66 spec.) were prevalent. The most frequently one form of deviation only was observed in one specimen of N. sympatrica. Nevertheless, the specimens simultaneously having several aberrations or anomalies were also found. 17 types of such combinations were observed, that counts 20.6% of all specimens with deviations. Symmetric deviations, namely the presence of two nude galeal setae (31 spec.), presence of 2 genualae on both legs I (4 spec.), presence of 2 AM (2 spec.) and symmetric reduction of scutal angles (1 spec.), sometimes cause troubles in diagnostics. The quarter of variance in N. sympatrica and in the species N. monticola Schluger et Davydov, 1967 formerly studied by the author turned out as almost identical. The specimens with deviations counted 14.5% of all studied specimens in the latter species. However, the structures of variance in these species is different. In N. monticola, the aberrations of humeral setae were dominant (71.6%) (Kharadov, Chirov, 2001), while in N. sympatrica, the aberrations of other structures were prevalent: genualae I (24.8%), AM (20.9%) and sternal setae (20.4%).     

Resting nymphal stages and the nature of individual development in trombiculid mites (Acariformes: Trombiculidae)

A morphological expression of quiescent nymphal instars (calyptostases) in trombiculid mites (Trombiculidae) is analysed. The analysis takes into consideration ontogenetic functions of different instars during the individual development of these mites, which includes an alternation of active and regressive quiescent instars. Cyclic start and blocking of different integumental differentiation programs in Trombiculidae is caused by the cyclic reduction and renewal of particular functions of respective instars, whereas internal organs of these mites do not transform significantly during the life cycle. The reduction of some functions, particularly the locomotion and feeding, in the quiescent nymphal instars with changed integumental function has an inevitable result in a complication and prolongation of the moulting process. The moulting of these instars begins at once after finishing the previous moulting cycle, i.e. it is realized in "automatic pattern" without the intermoult phase. In general, the organization of both quiescent proto- and tritonymph is the same, because it expresses concordant morphological correlations and synchronous reductions of certain functions and structures in these instars during the evolutionary process. At the same time, the trombiculid mites and, perhaps, other representatives of Parasitengona reveal the most generalized type of ontogenesis in Acariformes within the "alternating calyptostasy" phenomenon.     

Food preferences of tanypodinae larvae (Diptera: Chironomidae)

Analyses of the gut contents of larvae of three species of Tanypodinae, collected from four sites in north-east England and Scotland, invariably showed small particles (1–10 µm²), predominantly of detritus, to be more abundant than large particles (10–100 µm²), mostly algae and diatoms. No animal remains, such as carapaces, head capsules and chaetae, or vascular plant fragments were found in any of the intestines. In contrast, food-choice and growth experiments suggested that animal food (particularly Tubificidae and Chironominae larvae) was important in the tanypod diet. In the food-choice experiments, six different foods offered to small (4–7 mm long) and to large (8–12 mm long) larvae of Procladius choreus Meigen, were chosen in the following order: live Tubificidae spp.; detritus; live Chironominae larvae; a mixture of algae and diatoms; live Ostracoda spp.; and live Cladocera spp. However, when the results of the individual choices were examined, it was clear that the small larvae had consumed greater amounts of the small types of food (i.e. detritus, algae and diatoms) in comparison to the large larvae. In the growth experiment, those larvae of P. choreus reared solely on Tubificidae, detritus or Chironominae larvae thrived, while those fed on algae and diatoms, ostracods or cladocerans grew more slowly and fewer survived the experimental period.It was concluded that although Tanypodinae larvae are primarily predators with definite food preferences in ideal foraging conditions, in adverse conditions they will utilize a range of available stand-by foods, the most important of which appears to be detritus.     

Mouthparts and feeding of certain ground beetles (Goleoptera: Garabidae)

The feeding apparatus of 17 species of ground beetles (Carabidae), representing 17 genera and 12 tribes has been studied, and the observations correlated with gut content analyses and the ability of certain species to regurgitate pre-oral digestive fluids. The diverse feeding habits of species investigated include zoophagous fluid feeders, zoophagous fragmentary feeders and mixed feeders. In the last-named group, the food intake may be both fluid and fragmentary, and either of animal or of both animal and plant origin.     

Variability in Leptotrombidium europaeum and two new related chigger mite species (Acari: Trombiculidae) from Caucasus

Two new chigger mite species closely related to Leptotrombidium europaeum (Daniel et Brelih, 1959) are described from small mammals collected in Caucasus and Transcaucasia. L. alanicum sp. n. differs from L. europaeum in having shorter legs (TaIII = 61-81, Ip = 734-927 versus 72-90, and 855-1017), shorter scutal and idiosomal setae (D(min) = = 30-45, D(max) = 48-67, H = 59, PL = 58 versus 40-52, 54-69, 64, 63), slightly smaller scutum (AP = 25, SD = 47, PW = 89 versus 28, 50, 91), and more numerous idiosomal setae (87 versus 81). L. montanum sp. n. differs from L. europaeum in having more numerous idiosomal setae (102 versus 81), longer scutal and idiosomal setae (AM = 61, AL = 44, H = 68, D(max) = 66 versus 56, 41, 64, 62), thicker legs (TaW = 19 versus 18), and broader scutum (PW = 95 versus 91). Exact identification of both new species is possible only using classification functions constructed by means of discriminant analysis. These three Leptotrombidium species expose sympatric distribution in Daghestan (Eastern Caucasus). L. alanicum and L. montanum also occurred together in Krasnodar Territory (Western Caucasus). Each of these species includes a number of local geographical forms precisely distinguished from each other. Morphometric differences between L. alanicum and L. montanum agree with eco-geographical rules being previously found in chigger mites from other genera. However, differences between local forms of these species show other tendencies directed controversially in part. Therefore it is probable that interspecific differences in this case correspond to the variability which took place in the process of speciation.