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PubMed2003

Morphological and populational characteristics of hemocytes of Ornithodoros moubata nymphs during the ecdysial phase.

The ultrastructure and characteristics of hemocytes of argasid tick species, Ornithodoros moubata, during the ecdysdial phase are herein presented. Hemocyte classes/populations characterized based on their affinity with Giemsa stain and ultrastructural differences comprised the prohemocytes, nongranular cells (Nc), eosinophilic granular cells (Ec), basophilic granular cells (Bc), and unidentified cells. Significant changes/shift in the ratio of hemocyte classes/population was apparent in ticks before and after the ecdysial phase. The granule-scant basophilic granular cells (sBc) constituted the most abundant hemocyte population in the ecdysial phase. Nymphs in ecdysis showed increases in Nc and sBc and decrease in Ec, a phenomenon that was reversed in unengorged nymphs and adults ticks. The significant increase in total Bc population in ecdysis relative to nonengorged ticks clearly point to blastogenesis of Bc taking place during the ecdysial phase and Bc's important role in the process of tissue remodeling.

Animals↗

PubMed2004

Tickcidal effect of monoclonal antibodies against hemocytes, Om21, in an adult female tick, Ornithodoros moubata (Acari: Argasidae).

In the present study, monoclonal antibodies (mAbs) against adult Ornithodoros moubata hemocytes were established. Afterward, artificial feeding was performed to assess the tickcidal effect of fetal bovine serum meal containing each mAb. As a result, Om21 showed the strongest tickcidal effect on adult female O. moubata. The reactivity of various tick cells and organs, including the hemocyte, midgut, trachea, ovary, fat body, and muscle, to Om21 was then examined by an indirect immunofluorescent antibody test and by immunoelectron microscopy. Om21 reacted with not only hemocytes but also with fat body cells, epidermis, cuticle of the trachea, connective tissue of the muscle, and the basement membrane of the midgut, trachea, fat body, oocyte, and epidermis. These results suggest that Om21 passing through the midgut epithelium induced a tickcidal effect on hemocytes or various organs. However, the target of Om21 could not be identified in the present study. The antihemocyte mAb produced in this study, Om21, may be useful for the immunological control of ticks.

Animals↗

PubMed2005

Complement inhibitor of C5 activation from the soft tick Ornithodoros moubata.

Blood-feeding ticks must control C activation or be damaged by the host inflammatory response. We report the characterization and expression of a novel, relatively small, broad-acting C inhibitory protein (termed OmCI) from the soft tick Ornithodoros moubata. The native 17-kDa nonglycosylated protein inhibits both human and guinea pig classical and alternative C activation pathways. The IC50 values for each pathway were 12 and 27 nM, respectively, in hemolytic assays using human serum diluted 40-fold. The cDNA encodes a protein of 168 aa, including an 18-aa secretion signal sequence that is absent in the mature form. The inhibitor has 46% amino acid identity with moubatin, a platelet aggregation inhibitor also from O. moubata that is an outlying member of the lipocalin family. Native OmCI had no inhibitory effect on the addition of C8 and C9 to preformed C5b-C7 and C5b-C8 to form the membrane attack complex and no effect on the rate of C3a production by the C3 convertase enzymes C4bC2a, C3(H2O)Bb, or C3bBb. Both recombinant and native OmCI abolish production of C5a by human classical (C4bC3bC2a) and alternative (C3bC3bBb) C5 convertases. Addition of excess C5 but not C3 competes away the inhibitory activity of OmCI, indicating that OmCI targets C5 itself rather than inhibiting the C5 convertase C4bC3bC2a itself. Direct binding of OmCI to C5 was demonstrated by Western blotting and gel filtration chromatography using 125I-labeled proteins. OmCI is the first lipocalin family member shown to inhibit C and also the first natural inhibitor that specifically targets the C5 activation step.

Amino Acid Sequence↗

PubMed1993

Metabolism of [3H]-ecdysone in embryos and larvae of the tick Ornithodoros moubata.

The fate of [3H]-ecdysone ([3H]-E) was investigated in hanging drop cultures of embryos and larvae of the tick Ornithodoros moubata using HPLC. The hormone was metabolized more slowly during described periods of increasing endogenous ecdysteroid (ES) titers than during periods of low titers except for young embryos. Three different classes of metabolites were produced: 1) apolar products (AP) corresponding to C-22 fatty acid ester conjugates of E and, in some cases, of 20-hydroxyecdysone (20E), 2) unidentified polar products (PP) more polar than E, one peak of which had the same retention times as 20,26-dihydroxyecdysone, and finally, 3) 20E verified by comigration of cold standards on RP-18 and silica columns. Hydroxylation of E to 20E first became evident in cultures of 2 day old embryos and was present in all cultures of older animals. Highest production of free 20E occurred during increasing endogenous ES titers in embryos and during the ES peak in larvae. Conjugation of E to AP occurred in all stages investigated, but was more pronounced during periods of low endogenous ES titers, and may correspond to a detoxification mechanism. In contrast, PP were produced during high 20E production in embryos and during periods of high and decreasing endogenous titers in larvae.

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PubMed1986

Fine structure and development of the sperm of the tick Ornithodoros (Pavlovskyella) erraticus (Ixodoidea, Argasidae).

Sperm development in Ornithodoros (Pavlovskyella) erraticus includes the formation of subsurface cisternae in the primary spermatocytes, which divide meiotically to secondary spermatocytes and ultimately to spermatids. During spermiogenesis the spermatid undergo morphological transformation including polarization of the nucleus and subsurface cisternae, formation of a cisternal tube, and modification of the subsurface cisternae to cellular processes surrounded by cisternal vesicles. Further transformation occurs after spermatids are introduced into the female. The spermatid cisternal tube now invaginates to form an inner cord surrounded by an outer sheath. The invaginated inner cord elongates anteriorly as the outer sheath continues to invaginate posteriorly during spermiogenesis. With further elongation, the spermatid membrane ruptures anteriorly, leaving the inner cord exposed as the outer surface of the maturing sperm. Posteriorly, the original plasma membrane invaginates to form an acrosomal canal which becomes surrounded by an acrosome. The hemispherical anterior end of the mature sperm is covered with rows of projections separated from the remainder of the sperm by a row of fringed processes. Except for the posterior end, the rest of the sperm is covered by longitudinally distributed electron-dense cellular processes and an outer mat of more electron-lucent tubular elements. Mitochondria and bundles of microfibrils are found beneath the cellular processes. Microfibrils are suggested to be the principal contractile organelles responsible for sperm motility. Cellular processes appear to be the main external motile structures, while movements of tubular elements and fringed processes may also contribute to sperm motility.

Animals↗

PubMed1988

Fine structure of larval malpighian tubules and rectal sac in the tick Ornithodoros (Pavlovskyella) erraticus (Ixodoidea: Argasidae).

The fine structure of the Malpighian tubules (Mts) and rectal sac (rs) is described in the larval tick Ornithodoros (Pavlovskyella) erraticus before and after feeding up to molting. Mts consist of structurally different pyramidal and cuboidal cells along the entire length of the tubule. In unfed ticks, the two types of cell are characterized by apical microvilli and a few basal membrane infoldings. The abundant pyramidal cells contain glycogen particles, lipid droplets, lysosomelike structures, and rickettsialike microorganisms. After feeding but before molting, pyramidal cells loose glycogen particles and become very dense and dramatically reduced in size. These cells are possibly involved in the formation of guanine crystalloids as an excretory product. In contrast, cuboidal cells, filled with glycogen particles, free ribosomes, and mitochondria in unfed larvae, grow steadily after feeding; their cytoplasm becomes rich in lipid droplets in addition to showing an increase in glycogen particles. Lipid and glycogen could be the source of energy required for water and ion reabsorption in which cuboidal cells are probably involved. The paired-lobe rs consists of one type of cuboidal cells with basal membrane infoldings and a brush-border microvilli covered by a fuzzy coat of glycocalyx. These cells grow rapidly after feeding; they have functional features indicating extensive, selective reabsorption of essential components from excretory products.

Animals↗

PubMed1995

An inhibitor from the argasid tick Ornithodoros moubata of cell adhesion to collagen.

An inhibitor of the adhesion of human platelets to collagen was identified in soluble extracts of the soft tick Ornithodoros Moubata and purified by four chromatographic steps. The isolated inhibitor, TAI (Tick Adhesion Inhibitor), is a approximately 15-kDa protein that completely blocks the adhesion of platelets to collagen-coated microtiter plates with an IC50 of 8 nM. In the same concentration range it does not inhibit collagen-induced platelet aggregation or platelet adhesion to fibrinogen and has a partial inhibitory effect on platelet adhesion to fibronectin. TAI also blocks the adhesion of human endothelial cells to collagen, thus its inhibitory effect is not limited to platelets. TAI competes for the binding to platelets of a radiolabeled monoclonal antibody against the platelet glycoprotein Ia-IIa integrin complex. Based on its selective activity and small size, TAI is a promising new molecule for exploring cell-collagen interactions.

Animals↗

PubMed1999

Savignin, a potent thrombin inhibitor isolated from the salivary glands of the tick Ornithodoros savignyi (Acari: Argasidae).

A thrombin (E.C. 3.4.21.5) inhibitor, savignin, was isolated from the salivary glands of Ornithodoros savignyi by a combination of size exclusion, anion-exchange, and reversed-phase chromatography. The inhibitor has a molecular mass of 12,430.4 Da as determined by electrospray mass spectrometry. The behavior of savignin during anion-exchange chromatography indicated that it has an acidic pI. The available N-terminal sequence (residues 1-11) differed from that of ornithodorin with only one residue. Savignin inhibits thrombin-induced platelet aggregation, but has no effect on ADP- or collagen-induced aggregation. Kinetic studies indicated that savignin is a competitive, slow-, tight-binding inhibitor of alpha-thrombin (K(i) = 4.89 +/- 1.39 pM). Tight-binding kinetics showed that the inhibitor has a lower affinity for gamma-thrombin (K(i) = 22.3 +/- 5.9 nM). Plasmin, factor Xa, and trypsin are not inhibited by savignin.

Amino Acids↗

PubMed1995

Immunocytochemical mapping of FMRFamide-like peptides in the argasid tick Ornithodoros parkeri and the ixodid tick Dermacentor variabilis.

FMRFamide-like immunoreactivity was studied in the argasid tick Ornithodoros parkeri and the ixodid tick Dermacentor variabilis using immunocytochemistry based on the peroxidase-antiperoxidase method. FMRFamide-like immunoreactive cells are widely distributed in various regions of the tick synganglion including protocerebral, cheliceral, stomodeal, palpal, pedal I-IV, and opisthosomal regions in both species. However, there is one layer of immunoreactive cells located on the dorsal surface of the postoesophageal part of the synganglion that is found only in D. variabilis. Besides the immunoreactivity within the cell body and its axons, the neuropile and the neural lamella (the extracellular sheath of the synganglion) are rich in immunoreactive materials. Some coxal muscles are innervated by the FMRFamide-like immunoreactive processes of the nerve from the pedal ganglion.

Animals↗

PubMed1995

Respiratory metabolism of the soft tick, Ornithodoros turicata (Dugès).

The rate of oxygen consumption was investigated in fed larval, nymphal and adult Ornithodoros turicata ticks and in starved nymphal and adult ticks. Oxygen consumption rate of fed adult ticks increased with increasing temperature. The metabolic rate of adult ticks was affected by starvation whereby starved adult ticks showed a significantly lower oxygen consumption than their fed counterparts. The oxygen consumption rate of fed female ticks was significantly higher than that of fed males but, there was no significant difference between the oxygen consumption rates of starved female versus starved male ticks. Oxygen consumption of fed larvae was significantly greater than those of fed first through third instar nymphs. Fed and starved nymphal ticks as well as fed adult ticks ventilated continuously. In contrast, starved adults ventilated discontinuously. The ability to reduce metabolic rate, plus the capability to ventilate discontinuously allow O. turicata adults to cope with prolonged starvation.

Animals↗

PubMed1995

Identification of anticoagulant activities in the salivary glands of the soft tick, Ornithodoros savignyi.

Salivary gland extracts of the sand tampan, Ornithodoros savignyi, prolonged the activated partial thromboplastin time (APTT) and prothrombin time (PT) significantly in a concentration-dependent manner. There was also a pronounced inhibition of human activated factor Xa (fXa) by salivary gland extracts. The salivary gland extracts inhibited chromogenic assays specific for both fXa and thrombin. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of the salivary gland proteins followed by elution of specific areas or bands from a polyvinylidene difluoride (PVDF)-membrane, showed that various anticoagulant factors are present when screened by means of the APTT assay. The most active component was associated with a band of M(r) of 14 kDa. Partial purification of this component was achieved using isoelectric focusing (IEF) and size-exclusion high-performance liquid chromatography (HPLC).

Animals↗

PubMed1996

Isolation and characterization of an anticoagulant from the salivary glands of the tick, Ornithodoros savignyi (Acari: Argasidae).

An inhibitor of activated coagulation factor X (fXa) was isolated from salivary gland extracts prepared from Ornithodoros savignyi using a two-step procedure, involving reversed-phase high-performance liquid chromatography (RP-HPLC) and diethylaminoethyl (DEAE) ion-exchange chromatography. From its behaviour during DEAE chromatography it could be deduced that it possesses an acidic pI (approximately 4.6). Capillary zone electrophoresis (CZE) of the purified inhibitor showed it to be homogeneous. The molecular mass was determined as 12 kDa using capillary gel electrophoresis (CGE) and as 7183.4 using laser desorption mass spectrometry (LDMS). The N-terminal amino acid sequence (residues 1-12) was determined and found to share a 66% identity with tick anticoagulant peptide (TAP). The O. savignyi peptide is a slow, tight-binding inhibitor of fXa (Ki = 0.83 +/- 0.10 nM). The interaction of the fXa--inhibitor was found to be competitive and dependent on ionic strength. Preliminary investigations show that the inhibitor may be specific for fXa.

Animals↗

PubMed1991

Demonstration of regurgitation of gut content during blood meals of the tick Ornithodoros moubata. Possible role in the transmission of pathogenic agents.

Nymphs of Ornithodoros moubata were fed tritiated ecdysteroid. These ingested hormones are conjugated to fatty acyl esters that accumulate in the midgut (Connat et al. 1988). A few months later, the same ticks which had molted, were fed on physiological medium without radiolabel. At the issue of the blood meal, the nutritive medium contained an amount of radiolabel corresponding to 0.5% of the total labelling in the animal before the blood meal; this corresponded to 1.3% of the midgut content. These results demonstrate that in addition to transmission of parasites by saliva and coxal fluid (Burgdorfer 1951), transmission through regurgitation of the blood content in the gut could occur. An equivalent quantity of radiolabel was also emitted in the feces during and after the meal, but no conclusion about parasite transmission can be drawn from these "metabolic" results.

Animals↗

PubMed1991

Artificial feeding and successful reproduction in Ornithodoros moubata moubata (Murray, 1877) (Acarina: Argasidae).

The increasing demand for laboratory-reared argasid and ixodid ticks for research and control purposes makes it necessary to develop effective and standardized tick feeding methods without using live animals as hosts. The in vitro maintenance technique, described in this paper, has been used successfully for rearing Ornithodoros moubata moubata by feeding all nymphal and adult instars through Parafilm 'M' sealing film on heparinized bovine blood (fresh standard). The technique is based on a specially designed tick feeding apparatus with a capacity to feed 2000 first nymphal instars (N 1) or up to 200 adults at one time. For different 1-month-old instars feeding rates were between 80-100%. Using this feeding technique the subsequent egg production of female ticks was remarkably high, producing an average of 210.2 eggs per tick with a hatch rate of 96.72%. There was no overall difference in the reproductive capacity of 1-month-old female O. M. moubata fed on heparinized and haemolyzed bovine blood (kept deep-frozen), heparinized rat blood and defibrinated ovine blood when compared with those fed on heparinized bovine blood (fresh standard).

Animals↗

PubMed1991

Immunocytochemical localization of an insulin-like substance in the synganglion of the tick Ornithodoros parkeri (Acari: Argasidae).

Immunocytochemical staining based on the peroxidase-antiperoxidase method has shown that some neurosecretory cells (NSC) in the synganglion of the adult female tick Ornithodoros parkeri react with an antibody to bovine insulin. There are 18 regions of paraldehyde fuchsin-positive NSC of which three regions showed specific insulin-like immunoreactivity: anterolateral cheliceral, anteromedial stomodeal and posterior opisthosomal. Immunoreactivity can also be found in the extracellular surface of the neurilemma of the synganglion. This suggests a possible neurohemal site and release of neurohormone in a diffuse manner.

Animals↗

PubMed1988

Fine structure of the vagina, accessory glands, uterus, oviducts and Gene's organ in the unfed tick, Ornithodoros (Pavlovskyella) erraticus (Ixodoidea: Argasidae).

The fine structure of the reproductive system of the unfed female Ornithodoros (Pavloskyella) erraticus is described. The vagina consisting of vestibular (VV) and cervical (CV) regions is formed of a single epithelial layer lined with cuticle and surrounded by muscle layers. Epithelial cells of VV show no signs of activity, while those of CV have structural features of transporting epithelia. A pair of tubular accessory glands opening at the junction of the two vaginal regions consist of a layer of microvillate columnar cells containing rough endoplasmic reticulum; this is possibly involved in the formation of colloidal material which presumably coats the egg surface during its passage through the vagina. The bilobed uterus opening into the cervical vagina secretes material that is possibly responsible for the release of sperm from the endospermatophore. The paired oviducts joining the uterine lobes anteriorly have distinctive distal (DO) and proximal (PO) parts as well as 'ampullae' (AMP) at their junction. DO cells contain many lysosome-like structures which probably play a role in breaking down some sperms ascending to the ovary. Cells of AMP are packed with rickettsia-like Wolbachia, while those of PO contain a few granules. Gene's organ, present in female ticks only, consists of a stalk and two horns of epithelium lined internally by a cuticular layer. Bundles of microtubules are detected in the horn cell cytoplasm. This organ functions to waterproof the eggs with a waxy layer just before they are deposited.

Animals↗

PubMed1992

Effects of precocenes on vitellogenesis in the adult female tick, Ornithodoros moubata (Acari: Argasidae).

Vitellogenin (Vg) concentrations in the hemolymph and ovarian development were studied in Ornithodoros moubata after treatment with precocenes 1 (P1) and 2 (P2). Precocene was dissolved in acetone or DMSO and topically applied to the dorsal surface of ticks: (1) at adult ecdysis; (2) 24 h before engorgement; (3) immediately after engorgement; and (4) 24 h after engorgement. Subsequently, P1 and P2 were dissolved in olive oil and injected through the gonopore into the body cavity 24 h after engorgement. Vitellogenin concentration was measured on days 5 and 10 after engorgement and ovarian development was scored on day 10, 20 or 30. Oviposition was also recorded and the average weight of eggs laid by females was determined. No differences in concentration of Vg in the hemolymph occurred between the control ticks and ticks treated topically or by injection with P1 and P2. Precocene did not suppress the synthesis of Vg in O. moubata. However, oviposition was reduced in ticks that survived repeated treatment with high doses of P2 dissolved in acetone.

Animals↗

PubMed1985

The isolation of Meaban virus, a new Flavivirus from the seabird tick Ornithodoros (Alectorobius) maritimus in France.

Seven strains of a new Flavivirus, for which the name of Meaban virus is proposed, were isolated from the seabird tick Ornithodoros (A.) maritimus collected during 1981 and 1982 in nests of herring gulls (Larus argentatus) on islands of South Brittany, France. The new virus was compared serologically with 65 other flaviviruses including Tyuleniy virus and was found to be most closely related to, but different from Saumarez Reef virus, an agent previously isolated in Australia and Tasmania. Some general properties of Meaban virus are described and its antigenic relationships with other tick-borne flaviviruses associated with seabirds are discussed.

Animals↗