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Amblyomma variegatum (Acari: Ixodidae): mechanism and control of arbovirus secretion in tick saliva.

Saliva is considered to be the conduit by which pathogens are transmitted from blood-sucking arthropod vectors to their vertebrate hosts, but supporting evidence for this is fragmentary. To determine if Thogoto (THO) virus, a tick-borne member of the influenza virus family, is transmitted via tick saliva, and whether virus replication is a prerequisite for such transmission, two experimental conditions were compared: (1) "biological transmission" and (2) "mechanical transmission." In (1), THO virus was allowed to infect and replicate in a natural vector, Amblyomma variegatum: virus was detected in saliva collected from 3/22 (14%) ticks. In (2), virus was inoculated directly into the hemocoel with the drug used to induce salivation and saliva was collected immediately to preclude the possibility of virus replication: virus was detected in saliva collected from 31/170 (18%) ticks. The results demonstrate that THO virus is secreted in tick saliva and that virus can pass from the hemolymph to the salivary glands independently of viral replication within the tick. The comparatively low numbers of ticks that yielded virus-positive saliva samples together with the results from assays of serial saliva samples suggested that virus secretion may not be a continuous process during salivation. Ticks in which THO virus had established an infection showed an impaired secretory response compared with uninfected ticks and ticks used for mechanical transmission.

Analysis of Variance↗

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↗

Ultrastructure and Pathology of Microsporidium phytoseiuli n. sp. Infecting the Predatory Mite, Phytoseiulus persimilis Athias-Henriot (Acari: Phytoseiidae)

Ultrastructure and pathology of Microsporidium phytoseiuli n. sp. infecting the predatory mite Phytoseiulus persimilis Athias-Henriot is described using light and transmission electron microscopy. Infected mites showed no gross, external symptoms. All observed stages of the parasite had unpaired nuclei. Schizonts were commonly observed within nuclei of digestive cells of the ventriculus and within the cytoplasm of cells lining the cecal wall and in muscle tissue underlying it. Sporoblasts and spores occurred in the nuclei and cytoplasm of digestive cells within the ventriculus, in cortical regions of the sub- and supraesophageal ganglia, within the cecal wall and muscle tissue, and in parenchyma cells underlying the cuticle. Mature spores were also observed in developing eggs within gravid females. These were broad- to elongate-ovoid, measured 4.33 ± 0.35 x 1.27 ± 0.15 &mgr;m (electron micrographs), 5.37 ± 0.46 x 2.22 ± 0.17 &mgr;m (fixed and stained), and 5.88 ± 0.34 x 2.22 ± 0.19 &mgr;m (fresh) and had an isolfilar polar filament coiled 12 to 15 times within the posterior two-thirds. Within cells, individual spores appeared to be in direct contact with host cytoplasm, while groups of spores were infrequently observed within interfacial envelopes. Groups of 4, 8, to more than 16 spores were observed by light microscopy, while 8 was the maximum observed by electron microscopy. No spores were observed in Tetranychus urticae, a mite used as food during this study.

Journal Article↗

Birefringent Crystals and Abdominal Discoloration in the Predatory Mite Phytoseiulus persimilis (Acari: Phytoseiidae)

In response to grower complaints of poor performance of Phytoseiulus persimilis, mites from 14 commercial insectaries and research colonies were examined for pathogens. Some were found to have abdominal discolorations, manifested initially as two white stripes along the dorsal sides of the body within the Malpighian tubules. Advanced signs appeared as a large, centrally located, white spot or U-shaped discoloration in the distal opisthosoma within the rectum/anal atrium. White material often accumulated and hardened within the anus and formed a rectal plug that inhibited further excretion. Most affected mites were lethargic. Adults and immatures with abdominal discoloration contained numerous densely packed, birefringent, dumbbell-shaped entities. Though occasionally observed in the colon, they occurred most frequently within the Malpighian tubules and/or rectum and anal atrium. Dumbbells measured 2-4 &mgr;m long and contained prominent concentric rings. When observed by transmission electron microscopy, the entities lacked cellular organelles. Asymptomatic mites contained few or no such entities. Dumbbell-shaped inclusions were observed in P. persimilis from all sources examined. High levels of potassium, low levels of phosphorous and sulfur, and traces of chlorine were detected by energy-dispersive X-ray analysis. Guanine and uric acid, known nitrogenous wastes of arachnids, do not contain these elements. The chemical composition and structure indicate that the dumbbells are crystals. Both asymptomatic mites and those specimens exhibiting abdominal discoloration were examined for pathogens using light and transmission electron microscopy. Microsporidia, virus-like particles, and a rickettsia (genus Wolbachia) were observed in some mites but showed no correlation with white abdominal discoloration or associated crystal formation. Neither were pathogens always detected in symptomatic mites. Although birefringent crystals may be naturally occurring excretory products, the cause of white abdominal signs associated with crystal accumulation in P. persimilis is unknown. These signs indicate overall poor health.

Journal Article↗

The Effect of Pathogen Dosage on the Pathogenicity of Neozygites floridana (Zygomycetes: Entomophthorales) to Mononychellus tanajoa (Acari: Tetranychidae)

How the pathogen inoculum size affects the pathogenicity of Neozygites floridana to Mononychellus tanajoa was studied at 28°C, 12L:12D photoperiod and 90-100% RH. All inoculum sizes tested (one, two, four, six, and eight capilliconidia per mite) were lethal. Dead mites started to appear 48 hr after exposure to the conidia. Inoculum size significantly affected time to mortality. All mites exposed to six or eight capilliconidia died of mycosis 57.1 and 62.9 hr, respectively, after infection. At smaller doses, infection rates were lower, dead mites started to appear later, and time to mortality was significantly longer (between 68.7 and 89.4 hr). Most mites died within the first 6 hr of the dark phases of the photoperiod.

Journal Article↗

Disease prevalence and transmission of Microsporidium phytoseiuli infecting the predatory mite, Phytoseiulus persimilis (Acari: Phytoseiidae).

Isolated colonies of the predatory mite, Phytoseiulus persimilis, were used to gain information regarding prevalence and transmission of Microsporidium phytoseiuli. Two colonies of P. persimilis were reared on spider mite (Tetranychus urticae)-infested bean plants in isolated cages. Disease prevalence of predators from Colony 1 remained relatively low (between 0 and 15%) over 57 weeks of observation whereas disease prevalence of predators from Colony 2 increased over 3 months (from 12 to 100%). Disease prevalence among predators from Colony 1 had increased to 100% 2 months after weekly sampling had ceased for this colony and periodic sampling confirmed that disease prevalence among individuals of both colonies remained at 100%. Microsporidian spores were not detected in randomly chosen samples of T. urticae prey mites that were removed and examined biweekly during this period. Although numerous microsporidian spores were observed in smear preparations of fecal pellets examined by light microscopy, spores were not observed on leaf surfaces or predator feces when examined by SEM. The latter appeared as intact aggregates composed of numerous dumbbell-shaped crystals and it is unlikely that spores are liberated from intact fecal pellets onto leaf surfaces. Vertical transmission of M. phytoseiuli was 100%; horizontal transmission was low (14.3%) and occurred only when immature P. persimilis were permitted to develop in contact with infected immature and adult predators. The mean number of eggs produced per mated pair was highest when uninfected females were mated with uninfected males (63.2 eggs per mated pair). Although mean egg production decreased when one or both parents were infected, not all differences were significant. Male predatory mites did not contribute to infection of their progeny. Results suggest that routine examination of P. persimilis for microsporidian spores is essential for the management of M. phytoseiuli within P. persimilis colonies. Low disease prevalence and lack of obvious disease signs or symptoms, as in the case of M. phytoseiuli, increase the probability that these pathogens will escape notice unless individuals are routinely examined for pathogens.

Animals↗

Phylogenetic relationships within terrestrial mites (Acari: Prostigmata, Parasitengona) inferred from comparative DNA sequence analysis of the mitochondrial cytochrome oxidase subunit I gene.

Partial DNA and amino acid sequences translated from the mitochondrial cytochrome subunit I gene (408 bp) of 17 mite species have been used for analyzing the phylogenetic relationships within the terrestrial Parasitengona (Trombidia). Due to mutational saturation of the third codon position, only first and second codon positions and amino acid sequences were analyzed, applying neighbor-joining, maximum-parsimony, and maximum-likelihood tree-building methods. The reconstructed trees revealed similar topologies of taxa; however, the phylogenetic relationships could be convincingly resolved only within several trombidioid taxa. The proposed basic relationships within the Parasitengona, in particular those of Calyptostomatoidea, Smarididae, and Erythraeidae, were poorly supported in bootstrap tests. A comparison of the presented gene tree with a phylogenetic tree based upon traditional characters revealed only few contradictions in nodes only weakly supported by morphological data. The most astonishing result is the proposed early derivative position of Microtrombidiidae within the terrestrial Parasitengona.

Animals↗

An evaluation of strategic and threshold control measures against the Karoo paralysis tick, Ixodes rubicundus (Acari: Ixodidae) in South Africa.

Paralysis caused by feeding female Ixodes rubicundus ticks is a major problem in large areas of South Africa. As the life cycle of the tick extends over a period of 2 years, it was hypothesized that strategic treatment of sheep with an acaricide over a 2 year period, timed to kill most engorging females, should markedly lower the biotic potential of the tick. Two flocks of sheep grazing in separate paddocks known to be infested with I. rubicundus were treated either strategically or on a threshold basis (i.e. only when tick challenge exceeded a predetermined critical level in terms of paralysis) for a 2 year period. The tick burdens of untreated control sheep running with the two flocks were monitored over a 4 year period and their seasonal dynamics determined. The times at which peak infestations occurred were similar for both flocks of sheep, but significant differences in mean tick burdens between the two flocks were recorded. Tick numbers on sheep in the strategically treated flock did not decrease during the third and fourth years of the trial as was expected. Possible reasons for this were low stocking densities, especially during times of peak abundance of adults and the presence of wild hosts which maintained tick populations.

Animals↗

Hyalomma truncatum (Acari; Ixodidae): evidence for the inability of adult ticks to discriminate between colours.

Behavioural investigations into the perception and differentiation of coloured objects by unfed adult Hyalomma truncatum ticks revealed that silhouettes of blue, green, red and yellow colour, under illumination by a sun-simulating waveband spectrum, are perceived by the ticks and responded to equally by a directed response. Two green or dark grey rectangles each with a luminance contrast ratio of 5:1 against the white wall of the test arena in combination with an overlapping, equally sized dark grey or green target were consistently reached by ticks in a ratio of 2:1. Since the outer targets were occupied by the double number of ticks compared with the central silhouette this shows that the response is independent of the colour of the object. Investigations into target perception under monochromatic radiation of different wavelength ranges which were evenly adjusted in their irradiances revealed that ticks responded equally to a black target irradiated by blue, green, yellow and red light of wavelengths 428-472, 517-563, 549-591 and 606-654 nm, respectively. These results indicate the lack of true colour vision in H. truncatum.

Animals↗

Borrelia burgdorferi sensu lato in ticks (Acari: Ixodidae) from two different foci in Spain.

The prevalence of Borrelia burgdorferi sensu lato in several tick species was studied over a 2 year period in two ecologically different areas in Spain. One area was an endemic area for Lyme disease, with a number of autochthonous human cases and supported large populations of Ixodes ricinus on cattle and birds; the second area was characterized by the absence of I. ricinus together with the presence of foxes and their associated tick species. While I. ricinus was the main vector of B. burgdoreri in the endemic area (with a mean prevalence of 14% in adults and 51% in nymphs), adults of both Ixodes canisuga and Ixodes hexagonus had high rates of B. burgdorferi prevalence (30 and 28%, respectively) in the zone where I. ricinus was absent. Immatures of Ixodes frontalis were found to be carriers of the spirochete only in those zones where I. ricinus is present, suggesting evidence for reservoir competence in a tick-bird cycle.

Animals↗

A laboratory-based method to measure relative pesticide and spray oil efficacy against broad mite, Polyphagotarsonemus latus (Banks)(Acari: Tarsonemidae).

Six pesticides and two spray oils were tested against Polyphagotarsonemus latus. The chemicals were evaluated under laboratory conditions, requiring the development of a novel bioassay method, which is reported here. The pesticide toxicities fell into three distinct groups, namely abamectin, conventional pesticides and oils. The relative pesticide toxicities at the LC50 level were abamectin 4.9 x 10(-8) g ai 1(-1), endosulfan 1.1 x 10(-3) g ai 1(-1), fenpyroximate 2.3 x 10(-3) g ai 1(-1), pyridaben 4.1 x 10(-3) g ai 1(-1), tebufenpyrad 4.4 x 10(-3) g ai 1(-1), dicofol 4.5 x 10(-3) g ai 1(-1), petroleum spray oil 3.4 x 10(-1) g ai 1(-1) and canola oil 4.1 x 10-(1) g ai 1(-1). The calculation of the LC99.9 values allows for resistance monitoring in P. Latus and the suggested discriminating concentrations are abamectin 1.0 x 10(-4) g ai 1(-1); endosulfan, pyridaben and dicofol 1.0 x 10(-1) g ai 1(-1) fenpyroximate and tebufenpyrad 5.0 x 10(-1) g ai 1(-1).

Animals↗

Acquired resistance in dogs to repeated infestation with Ixodes scapularis (Acari: Ixodidae) reduces tick viability and reproductive success.

The purpose of this study was to determine whether dogs develop acquired resistance to adult Ixodes scapularis infestation in an experimental model. Five dogs were each infested with ten mating pairs of ticks every week for 7 consecutive weeks, another five dogs were each infested with ten mating pairs once every 2 weeks for 10 weeks and four dogs served as controls not exposed to ticks. All ticks were allowed to feed to repletion and were collected only after dropping from the host. Several variables were measured to determine the extent of blood feeding success. Regression analysis indicated that the engorgement success, survival and mean tick engorgement weight declined with repeated infestation in both groups of dogs (p < 0.05). Tick oviposition as well as the F1 viability declined with each successive infestation in both groups. These results suggest that repeated infestation with I. scapularis elicits a protective immune response against tick feeding and could serve as a limiting factor in the spread and transmission of Borrelia burgdorferi.

Animals↗

Efficacy of pheromone-acaricide-impregnated tail-tag decoys for controlling the bont tick, Amblyomma hebraeum (Acari: Ixodidae), on cattle in Zimbabwe.

A large-scale field test using pheromone-acaricide-impregnated plastic tail-tag decoys demonstrated excellent efficacy of these devices for control of the bont tick, Amblyomma hebraeum, on cattle in Zimbabwe. The tail tags were impregnated with a mixture containing o-nitrophenol, methyl salicylate, 2,6-dichlorophenol and phenylacetaldehyde and one of three different acaricides (cyfluthrin, flumethrin or alphacypermethrin). o-Nitrophenol and methyl salicylate are components of the A. hebraeum attraction-aggregation-attachment pheromone, while 2,6-dichlorophenol and phenylacetaldehyde are proven attractants for this tick. Both o-nitrophenol and methyl salicylate were lost gradually from the tags over 12 and 14 week periods, respectively. In field trials, tick counts were compared between cattle that received tail tags either impregnated with pheromone mixture alone, cyfluthrin and pheromone mixture, flumethrin and pheromone mixture, alphacypermethrin and pheromone mixture or were left untreated. During the first 3 month trial period, control of adult bont ticks was 94.9% with cyfluthrin tail tags and 87.5% with flumethrin tail tags. In general, there was no significant difference in bont tick numbers on cattle without tags and those with tail tags containing pheromone only. When the trial was repeated for another 3 month period, control of bont ticks with tail tags containing cyfluthrin and flumethrin was 99.3 and 95.1%, respectively. However, control of bont ticks using alphacypermethrin was only 79.2%. Overall, retention of tail tags was excellent although some loss was encountered during the rainy season. In addition to controlling bont ticks, the tail tags provided moderate control of other tick species (Rhipicephalus evertsi evertsi, Rhipicephalus zambeziensis and Hyalomma spp.) simultaneously infesting cattle in the trials.

Animals↗

Resistance and cross-resistance in rabbits to adults of three species of African ticks (Acari: Ixodidae).

Resistance to Rhipicephalus appendiculatus, Amblyomma variegatum and Amblyomma hebraeum was investigated in the laboratory by infesting rabbits with adults of each of the three species followed by homospecific or heterospecific secondary infestations. Significantly lower female engorged weights and egg mass weights were taken as evidence of protective immunity. Following a single infestation with adults, rabbits developed homospecific protective immunity (resistance) to only R. appendiculatus and A. hebraeum; primary infestation with A. variegatum did not protect against secondary infestation with the same species. There was no cross-resistance (heterospecific protective immunity) between the species except for one-way protection between R. appendiculatus and A. variegatum; primary infestation with R. appendiculatus protected against secondary infestation with A. variegatum, but not vice versa. The results from ELISA did not indicate any correlation between serum antibodies to soluble antigens from salivary gland extracts and protective immunity. Post-infection sera from rabbits infested with each of the three species reacted strongly to their respective salivary gland extracts. Despite the high reactivity of A. variegatum serum with salivary gland antigens from all three species, A. variegatum-infested rabbits did not show any homospecific or heterospecific-immunity; on the other hand, although R. appendiculatus serum did not react positively to A. variegatum antigens, infestation with R. appendiculatus protected against a subsequent A. variegatum infestation.

Animals↗

A population model for Dermanyssus gallinae (Acari: Dermanyssidae).

A model based on a time-varying distributed delay with attrition was developed for simulating the population dynamics of the chicken mite Dermanyssus gallinae (DeGeer, 1778). The model was parametrised according to cohort life table studies conducted under constant temperature conditions and was validated with two independent data set obtained in an experimental poultry house. The predicted mite densities and the stage-structure of the population corresponded to the values observed during experimental periods of 32, and 11 weeks, respectively. Temperature and a poultry house-specific density effect were determined to control the population development.

Animals↗

Sites of attachment and density assessment of female Ixodes ricinus (acari: ixodidae) on dairy cows.

A sample of 110 lactating dairy cows was examined to describe the attachment sites of female Ixodes ricinus ticks and to assess whether counts on one or two body areas can predict infestation and its level. Seventy per cent of the cows were infested by I. ricinus and the average tick burden on the infested cows was 15, ranging from 1 to 136. Preferred attachment sites were the axilla, udder/groin, neck, dewlap and flank. Udder/groin and axilla carried respectively 35.3% and 44.1% of the total tick burden. To predict infestation (yes/no), sensitivity of counts on axilla, on udder/groin and on both areas was respectively, 76.6%, 80.6% and 94.8%. Total burden prediction equations based on count on axilla and udder/groin are presented.

Animals↗