Esterase and lipase in camel tick Hyalomma dromedarii (Acari: Ixodidae) during embryogenesis

Esterase and lipase activity showed significant changes during embryogenesis of camel tick Hyalomma dromedarii. From the elution profile of chromatography on DEAE-cellulose, six forms of H. dromedarii esterase (El to EVI) can be distinguished. Esterase EIII was purified to homogeneity after chromatography on Sepharose 6B. The molecular mass of esterase EIII was 45 kDa for the native enzyme and represented a monomer of 45 kDa by SDS-PAGE. Esterase EIII had an acidic pI at 5.3. Lipase activity was detected in the same DEAE-cellulose peaks (LI to LVI) of H. dromedarii esterases. The highest lipase activity was exhibited by lipase LIII. Esterase EIII and lipase LIII were compared with respect to Michaelis constant, substrate specificity, temperature optimum, heat stability, pH optimum, effect of metal ions and inhibitors. This study suggests that H. dromedarii lipolytic enzymes may play a central role in the interconversion of lipovitellins during embryogenesis.     

Ultrastructure of the nymphal integument in the camel tick Hyalomma (Hyalomma) dromedarii (Ixodoidea: Ixodidae)

The ultrastructure of the nymphal integument in the ixodid tick Hyalomma (Hyalomma) dromedarii is compared for stages of development during and after feeding, and up to the first step of molting, apolysis. The integument comprises a cuticular layer and underlying epidermal cells. The body cuticle, which consists of both sclerotized and non-sclerotized parts, is divided into an outer, thin epicuticle, and an inner, thick, fibrillar procuticle. Pore canals in the procuticle are continuous with wax canals which traverse the epicuticle. As feeding progresses, the parallel, extensible epicuticular folds disappear due to the gut filling with ingested blood. The procuticular zone, however, becomes subdivided into an exocuticle, similar to the previously seen procuticle, and a lamellate endocuticle. Pore canals lose their parallel pattern and appear to have become deformed by stretching of the cuticle. The flat epidermal cells grow asynchronously during feeding; their cytoplasm becomes packed with well-developed rough endoplasmic reticulum (RER), while the cell apices project long microvilli extending deep into the procuticle. The RER undergoes ultrastructural changes indicating synthetic activity. Dense material released through the microvilli may serve to lyse the endocuticle and thus cause separation of the cuticle from the epidermis during apolysis. The lysed area, the exuvial cavity, is filled with lysed components which are probably withdrawn by endocytosis into the apical coated vesicles which appear in epidermal cells. Two types of integumental glands, which may participate in wax production, are observed in this study. The ultrastructure of their previously undescribed cuticular ducts is described, in addition to other hypodermal structures including epidermis-muscle attachments and sensory receptors.     

Fine structure of the Gene's organ in the camel tick Hyalomma (Hyalomma) dromedarii (Ixodoidea: Ixodidae)

Gene's organ of the camel tick Hyalomma (Hyalomma) dromedarii is located in the anterodorsal region of the body cavity ventrad to the scutum. It consists of a short stalk, dividing posteriorly into 2 pairs of horns and then into tubular glands. In unfed ticks, the epithelial layer of both the stalk and horns is lined internally by 2 cuticular layers; an inner, thin, greatly folded, dense layer surrounds the organ main lumen, and an outer, thick, slightly folded, less dense layer abuts the cell apices. Only the inner cuticular layer extends into the horn posterior region and appears perforated with numerous pore canals and covered with fine, cuticular projections. The horn and tubular glands epithelium is structurally consistent with a secretory function that apparently increases as feeding progresses. During oviposition, the inner cuticular layer unfolds and inflates into a pair of balloonlike structures that evert through the organ external aperture to receive and manipulate each egg as it is laid, coating it with a waxy layer that prevents desiccation. The fine cuticular projections may have a function in gripping the eggs as they leave the vagina. This organ appears to be everted by hydrostatic pressure from the hemolymph and is retracted by muscles.     

Fine structure of muscles in the tickHyalomma (Hyalomma) dromedarii (Ixodoidea: Ixodidae)

Skeletal and visceral muscles are distinguished in the unfed nymphHyalomma (Hyalomma) dromedarii according to position, structure and function. The skeletal muscles include the capitulum, dorsoventral and leg oblique muscles. Their muscle fibres have the striated pattern of successive sarcomeres whose thick myosin filaments are surrounded by orbitals of up to 12 thin actin filaments. The cell membrane invaginates into tubular system (T) extending deeply into the sarcoplasm and closely associated to cisternae of sarcoplasmic reticulum (SR). The T and SR forming two-membered dyads are considered to be the main route of calcium ions whose movements are synchronized with the motor impulse to control contraction and relaxation in most muscles. Two types of skeletal muscle fibres are recognized, and are suggested as representing different physiological phases.In the visceral-muscle fibres investing tick internal organs, the actin and myosin filaments are slightly interrupted, and the T and SR are well demonstrated. Both skeletal and visceral muscles are invaginated by tracheoles and innervated by nerve-axons containing synaptic vesicles.     

Fine structure of the sight organs in the tickHyalomma (Hyalomma) dromedarii (ixodoidea: Ixodidae)

The paired eyes in adultsHyalomma (Hyalomma) dromedarii consists of a cuticular, transparent, convex lens extending downward as an inner, hemispherical, semitransparent projection, and underlying group of photoreceptor cells. The lateral cell cytoplasm consists mainly of interrupted cisternae of rough and smooth endoplasmic reticulum, while the cell middle side is packed with rhabdomeric microvilli. The cell basal regions become function axons forming the optic nerve. These cells are adapted to distinguish between light and darkness.     

Hyalomma dromedarii (Acari: Ixodoidea: Ixodidae): Central and peripheral nervous system anatomy

TheHyalommadromedarii central nervous system, the synganglion, is an integrated nerve mass concentrated around the esophagus and formed by fusion of a small anterodorsal supraesophageal part an a large posteroventral subesophageal part. The supraesophageal part consists of the protocerebrum including a pair of optic ganglia, a pair of cheliceral ganglia, a pair of pedipalpal ganglia, and the stomodeal pons. The subesophageal part includes four paired pedal ganglia and the complex opisthosomatic ganglion. The peripheral nervous system includes the following pairs of nerves: optic, cheliceral, pedipalpal, primary and accessory (histologically traced); also unpaired pharyngeal and recurrent nerves, four pairs of pedal nerve trunks, each with a hemal branch, and two pairs of opisthosomatic nerves. Each peripheral nerve is traced distally to the innervation site. The salivary glands are innervated anteriorly by branches of the pedipalpal nerve and medially by branches of the hemal nerves associated with the third pedal nerves.Reprint request should be sent to: Medical Zoology Department, NAMRU-3, Fleet Post Office, New York 09527, U.S.A.     

Traffic of the tick embryo basic protein during embryongenesis of the camel tick Hyalomma dromedarii (Acari: Ixodidae)

The tick embryo basic protein (TEBP) is present in the nucleus as a counterpart of histones at early embryonic stages of the tick Hyalomma dromedarii. The sharp drop in the TEBP nuclear level and elimination of the N-terminal dipeptide (leucine–serine) between days 12 and 15 after oviposition suggested the transport of TEBP to the cytoplasm for protein turnover. The traffic of TEBP during tick embryogenesis was examined. The level of TEBP was detected in the cytoplasm from the different embryonic stages by the established enzyme-linked immunosorbent assay (ELISA) and confirmed by immunoblotting. At day 12, a 2-fold increase in the cytoplasmic TEBP level coincided with its decrease in the nucleus. This result indicates that TEBP starts to leave the nucleus for the cytoplasm at day 12. The changes in the cytoplasmic leucine aminopeptidase (LAP)-specific activity were followed during tick embryogenesis. The LAP activity started to increase at day 12 and reached its maximum level at day 21. The enzyme displayed an optimum pH between 7.5 and 8.8 and a K_m value of 0.5 M for leucine-p-nitroanilide. The involvement of the exopeptidase activity in the TEBP turnover after its translocation to the cytoplasm is discussed.     

Effect of blood meal and mating on the genital tract ultrastructure in the female camel tickHyalomma (Hyalomma) dromedarii (Ixodoidea: Ixodidae)

The genital tract ultrastructure in the femaleHyalomma (Hyalomma) dromedarii is described during feeding and mating and up to oviposition. The vagina, consisting of vestibular (VV) and cervical (CV) regions, is formed of an epithelium lined internally with a folded cuticular layer and surrounded externally by muscle layers. These facilitate the passage of endospermatophores containing sperm into the receptaculum seminis (RS), and ova to the exterior. A pair of tubular accessory glands (AG) opening at the junction of VV and CV consist of an epithelial layer of undifferentiated cells. As feeding progresses, these cells synthesise their granular secretion which lubricates the egg surface during its passage through VV. The RS, opening anteriorly into the CV, consists of cuboidal cells lined with a thin cuticular layer. These cells become rich in glycogen and lipid vacuoles, possibly acting as a source of energy for various cell activities including granule synthesis and exocytosis. The granules discharge their contents into intercellular spaces distributed throughout the RS wall and communicate with the main lumen via narrow channels. The cell secretion may dissolve the endospermatophore wall to release sperm, while their lysosome-like structures may function in the breakdown of endospermatophoric material taken up by pinocytosis. The connecting tube (CT) opens into the CV anterodorsally and leads into the common oviduct (COV) posteriorly. It consists of an epithelium lined by a closely-adhering cuticular layer, giving the tube lumen the appearance of an undulate labyrinth with a complicated configuration. No secretory activity in the CT has been observed before, during, or after feeding. The paired, non-cuticular oviducts, extending from the ovary and fusing anteriorly to form COV, consist of an epithelium poor in cell organelles. At the final stages of feeding the cell cytoplasm contains large, phagosomal vacuoles penetrated by sperms, in addition to micropinocytotic vesicles which serve to break down the seminal fluid and other materials. The basal membrane is infolded giving characteristic features, which increase dramatically during oviposition, of epithelia involved in ion and water transport. The oviducal secretion may function as a tanning agent to harden the egg-shell and may also act as a lubricant for egg passage.     

Phylogeography of Hyalomma (Euhyalomma) lusitanicum (Acarina,Parasitiformes, Ixodidae) in Andalusia based on mitochondrial cytochrome oxidase I gene

The genetic population structure relationships of Hyalomma (Euhyalomma) lusitanicum in Andalusia (the south of the Iberian Peninsula) were examined using mtDNA sequence data from 887 bp of cytochrome oxidase subunit I (COI) gene. The sequence for the COI region was determined for 84 individuals collected in several localities of Andalusia, and 10 for other localities (i.e., five from Toledo, central Iberian Peninsula, four from Sicily (Italy) and one from Canary Island). Seventeen haplotypes were detected, including 27 polymorphic sites. The number of amino acid substitutions per site from mean diversity calculations for the entire population was 0.017. AMOVA analysis revealed a low gene flow that characterises the genetic population structure of this species in South Iberian Peninsula, with a haplotype diversity (h) value of 0.815. No geographically induced differentiation was observed, and separate evolutionary units were not detected. Our results indicate low genetic diversity across the geographical range of H. lusitanicum tick in Andalusia. Our data do not show any genetic discontinuity between the tick populations studied, including specimens from Canary Island and Sicily (Italy).

     

Isolation and properties of two forms of thrombin inhibitor from the nymphs of the camel tick Hyalomma dromedarii (Acari: Ixodidae)

Two forms of the nymphal thrombin inhibitors (NTI) 3.2 kDaand 14.9 kDa were purified by chromatography on CM-cellulose,Sephacryl S-300 and Sephadex G-50 columns and designated NTI-1 and NTI-2respectively. The NTI-2 turned out to be homogenous monomeric protein in bothnative-PAGE and denatured SDS-PAGE with M(r) value of 14.9 kDaapproximately and its pI value ranged from 7.2 to 7.5. The NTI-1 and NTI-2displayed anticoagulant activity since they prolonged both the activatedpartialthromboplastin time (APTT) and the prothrombin time (PT) of the camel plasma ina concentration-dependent manner. The potency of NTI-1 toward thrombin was5-fold higher than that toward FXa, while NTI-2 was 3-fold active toward FXathan thrombin. However, both of them did not inhibit any of the other examinedproteases. The types of inhibition of thrombin by NTI-1 and NTI-2 were non-competitive and competitive with inhibition constants (Ki) values of 11.7M and 211 nM respectively. One binding site wasdeduced on thrombin for each inhibitor.     

Towards a diagnostic view of Hyalomma (Hyalomma) aegyptium (Acari,Ixodidae)

The diagnostic characters of larval, nymphal and adult Hyalomma aegyptium (L., 1758) based on specimens from the territory of most part of the area are given. In the diagnoses of immature stages, was used the characters, which were formerly tested by the author for diagnostics of other Hyalomma species occurring in the former USSR. Commonly used characters and those which were revealed by the author as useful for the majority of Euhyalomma Filippova, 1984 and Hyalommina Schulze, 1919 species, have been used in the diagnoses of male and female. Differential diagnosis of Hyalomma aegyptium. Female: genital orifice as wide arch with straight posterior margin (fig. 2, 5); vestibular part of vagina funnel-like, greatly swollen (fig. 2, 5); setae of alloscutum stick-like, tapering in apical one (figs 2, 3, 4); second segment of palps with proximal narrowing (figs 3, 1, 2); spurs of coxae I widely separated, triangular, wide, subequal in size (fig. 3, 5). Male: any grooves of conscutum absent, except short and pit-like cervical ones (fig. 4); punctation sparce and impressive (fig. 4); adenal shields short and wide, without inner branch, posterior part widened, anteromedian margin straight (fig. 5, 4); spurs of coxae I widely separated, triangular, wide, subequal in size (fig. 6, 5). Nymph: posteromedian setae of alloscutum stick-like and, as a rule, with indented apices (fig. 7, 2); spurs of coxae I large, median spur as equilateral triangle and shorter than lateral one (fig. 7, 8); spurs of coxae II-IV well developed, with acute apices (fig. 7, 8). Larva: posterior part of scutum (behind the eyes) heavy elongated, its apex straight, postero-lateral incisions weakly developed (fig. 8, 1); spurs of coxae I as equilateral triangle in shape and with rounded apices, spurs of coxae II-III very large (fig. 8, 5).     

Hyalomma (Hyalommina) arabica: nymphal and larval identity and spiny mouse hosts in Saudi Arabia (Acarina: Ixodoidea: Ixodidae)

Nymphal and larval Hyalomma (Hyalommina) arabica Pegram, Hoogstraal, and Wassef, 1982, described herein, closely resemble those of Hyalomma (Hyalommina) rhipicephaloides Neumann, differing chiefly in body size (both stages), nymphal basis capituli and scutal proportional dimensions, and distinctness of larval coxal spurs. Females of these species are also structurally similar but males differ in major critical characters. Nymphs and larvae of both species parasitize the spiny mouse (Acomys spp.), but immatures of the third African- Arabian species of this subgenus, Hyalomma (Hyalommina) punt Hoogstraal, Kaiser, and Pedersen, are unknown. Hyalomma (Hyalommina) arabica occurs in valleys and hills of western Saudi Arabia and western Yemen; H. (H.) rhipicephaloides in the Red Sea and Dead Sea areas; and H. (H.) punt in northeastern Somalia and eastern Ethiopia. The ibex (Capra ibex nubiana Cuvier) is probably the original host of adult H. (H.) arabica and H. (H.) rhipicephaloides; the related domestic goat is an important host of adults of the 3 species, which also parasitize domestic sheep. Gazelles are recorded hosts of adults of H. (H.) rhipicephaloides and H. (H.) punt and the latter is also recorded from goats, sheep, camels and cattle.     

Ultrastructural characterization of cultured embryonic cells of Rhipicephalus appendiculatus neumann (Acarina : Ixodidae)

Cultured embryonic cells of Rhipicephalus appendiculatus (Acarina : Ixodidae) have been studied by electron microscope. Two major cell types, an epithelial-like cell and the fibroblast-type cell, are described. A possible third cell type, a muscle cell, which occurs rarely in the cell culture, is also described. The different phases of development, namely the growth, maturation, and aging of the cells are described and discussed.