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Molecular cloning and comparative analysis of fibrinogen-related proteins from the soft tick Ornithodoros moubata and the hard tick Ixodes ricinus.

Among disease-vectors, the evolution of the tick innate immune system is still lagging when compared to insects. Such an investigation, which was initiated, by first cloning and sequencing lectins associated in the innate immunity of invertebrates and having fibrinogen related domains, helped in the sequencing of cDNA encoding for OMFREP from the soft tick, Ornithodoros moubata. Also obtained were Ixoderin A and Ixoderin B cDNA sequences from the hard tick Ixodes ricinus. Tissue-specific expression of OMFREP showed that it was present primarily in the hemocytes and salivary glands. Ixoderin A besides sharing a similar expression profile was also expressed in the midgut. Both showed significantly high homology to the lectin Dorin M, from O. moubata. Further, phylogenetic comparisons between these molecules of the soft and hard ticks showed their relatedness to Tachylectins 5A and 5B, involved in the innate immunity of Tachypleus tridentatus and ficolins from both vertebrates and invertebrates. Ixoderin B showing tissue-specific expression only in the salivary glands and the sequence displaying certain motif differences in homology point towards a possible function different from the other two molecules. This is the first report of lectin-like sequences, with a fibrinogen-domain, from the hard tick I. ricinus and a preliminary phylogenetic study of these tick sequences with related fibrinogen-domain containing sequences highlights a possible role for them in the innate immunity of the ticks.

Amino Acid Sequence↗

A serine proteinase inhibitor (serpin) from ixodid tick Haemaphysalis longicornis; cloning and preliminary assessment of its suitability as a candidate for a tick vaccine.

Rapid amplification of the cDNA ends (RACE) and primers designed based on a conserved serpin amino acid motif (NAVYFKG) were used to clone a 1350bp cDNA which encodes a 378 polypeptide with high sequence similarity to several known serpins. We have named this gene as Haemaphysalis longicornis serpin-1 (HLS1). Northern blotting and reverse transcription (RT)-PCR analysis of total RNA from unfed or partially fed whole ticks as well as dissected tick organs revealed that transcription of HLS1 mRNA was induced by blood meal feeding during the slow feeding phase (24-48 h post-attachment) only in the tick midguts. Vaccination of rabbits with recombinant HLS1 (rHLS1) expressed in Escherichia coli resulted in 43.9 and 11.2% mortality of nymph and adult ticks which were fed on immunized rabbits. Polyclonal rabbit antibodies to tick saliva did not react with rHLS1, suggesting that native HLS1 was not secreted into the host during tick feeding. rHLS1 could be a potential candidate for a cocktail anti-tick vaccine.

Amino Acid Sequence↗

Antibody levels to recombinant tick calreticulin increase in humans after exposure to Ixodes scapularis (Say) and are correlated with tick engorgement indices.

The antibody responses of subjects who presented with a definite Ixodes scapularis (Say) tick bite were measured to determine the utility of the antibody response against recombinant tick calreticulin (rTC) as a biologic marker of tick exposure. Subjects bitten by I. scapularis evidenced an increase in anti-rTC antibody levels between visit 1 and visit 2 from 24.3 to 27.1 ng/microl serum (n = 88, p = 0.003), and levels remained elevated at visit 3 (p = 0.005). These anti-rTC antibody levels during visits 2 and 3 were significantly higher than those in four non-exposed controls. Tick engorgement indices, measured on the biting ticks, were found to be correlated with anti-rTC antibody levels (e.g., for visit 3: Pearson's r = 0.357, p = 0.001). Tick engorgement index (TEI), ratio of body length to scutal width, was identified to be the only independent predictor of anti-rTC antibody levels in linear regression models. Logistic regression revealed that a bite from an I. scapularis tick that became engorged (TEI >3.4) was a risk factor for anti-rTC antibody seropositivity (adjusted odds ratio for age and bite location = 7.4 (95% confidence interval 2.1-26.4)). The anti-rTC antibody test had a sensitivity of 0.50 and a specificity of 0.86 for a bite from I. scapularis that became engorged. Immunoblotting revealed that subjects made a specific anti-rTC antibody response.

Adult↗

Longitudinal analysis of tick densities and Borrelia, Anaplasma, and Ehrlichia infections of Ixodes ricinus ticks in different habitat areas in The Netherlands.

From 2000 to 2004, ticks were collected by dragging a blanket in four habitat areas in The Netherlands: dunes, heather, forest, and a city park. Tick densities were calculated, and infection with Borrelia burgdorferi and Anaplasma and Ehrlichia species was investigated by reverse line blot analysis. The lowest tick density was observed in the heather area (1 to 8/100 m2). In the oak forest and city park, the tick densities ranged from 26 to 45/100 m2. The highest tick density was found in the dune area (139 to 551/100 m2). The infection rates varied significantly for the four study areas and years, ranging from 0.8 to 11. 5% for Borrelia spp. and 1 to 16% for Ehrlichia or Anaplasma (Ehrlichia/Anaplasma) spp. Borrelia infection rates were highest in the dunes, followed by the forest, the city park, and heather area. In contrast, Ehrlichia/Anaplasma was found most often in the forest and less often in the city park. The following Borrelia species were found: Borrelia sensu lato strains not identified to the species level (2.5%), B. afzelii (2.5%), B. valaisiana (0.9%), B. burgdorferi sensu stricto (0.13%), and B. garinii (0.13%). For Ehrlichia/Anaplasma species, Ehrlichia and Anaplasma spp. not identified to the species level (2.5%), Anaplasma schotti variant (3.5%), Anaplasma phagocytophilum variant (0.3%), and Ehrlichia canis (0.19%) were found. E. canis is reported for the first time in ticks in The Netherlands in this study. Borrelia lusitaniae, Ehrlichia chaffeensis, and the human granylocytic anaplasmosis agent were not detected. About 1.6% of the ticks were infected with both Borrelia and Ehrlichia/Anaplasma, which was higher than the frequency predicted from the individual infection rates, suggesting hosts with multiple infections or a possible selective advantage of coinfection.

Anaplasma↗

A new tick-borne encephalitis-like virus infecting New England deer ticks, Ixodes dammini.

To determine if eastern North American Ixodes dammini, like related ticks in Eurasia, maintain tick-borne encephalitis group viruses, we analyzed ticks collected from sites where the agent of Lyme disease is zoonotic. Two viral isolates were obtained by inoculating mice with homogenates from tick salivary glands. The virus, which was described by reverse transcriptase polymerase chain reaction and direct sequencing of the amplification products, was similar to, but distinct from, Powassan virus and is provisionally named "deer tick virus." Enzootic tick-borne encephalitis group viruses accompany the agents of Lyme disease, babesiosis, and granulocytic ehrlichiosis in a Holarctic assemblage of emergent deer tick pathogens.

Animals↗

Persistence of tick-borne virus in the presence of multiple host species: tick reservoirs and parasite mediated competition.

Tick-borne viruses in tropical and temperate parts of the world have a significant impact on human, livestock and wildlife hosts both directly, through mortality/morbidity, and economically. Since the ticks have multiple life stages and can utilize a large range of host species our understanding of the dynamics of these infections is often not clear. In this paper we consider the impact of a population which is a tick host but non-viraemic on one which is both a tick host and viraemic. We present two simple deterministic models and use joint threshold density curves to illustrate the basic reproductive ratios of both the ticks and the virus. We find that the non-viraemic hosts can have considerable impact on the viraemic host. Either they amplify the tick population and cause the virus to persist, or they dilute the infection and cause it to die out. A general model framework is presented here but a special case of this model describes the red grouse-hare-Louping-ill system.

Animals↗

Immunisation of rabbits against the brown ear tick, Rhipicephalus appendiculatus using tick haemolymph.

Tick naive rabbits were immunised with haemolymph components from partially fed Rhipicephalus appendiculatus adult ticks and subsequently challenged with all the developmental instars of the tick. The results obtained showed that the rabbits were rendered resistant to all the instars of the tick. However, the resistance was more pronounced in adult ticks than in the immature stages. The resistance was manifested as a reduction in the number of ticks that fed successfully to engorgement, reduced engorgement weights and reduced fecundities. Re challenging the resistant experimental rabbits with all the developmental instar stages of Rhipicephalus appendiculatus showed that resistance was maintained in subsequent infestations.

Animals↗

Ticks and tick-borne bovine piroplasmosis in the Southmountain pasture of Hunan Province, China.

Ticks and tick-borne bovine piroplasmosis is one of the most serious diseases affecting cattle bred in the Southmountain pasture of Hunan province. An investigation carried out from 1992 to 1995 showed that Theileria sergenti was found in the blood of disease-stricken cows. Ticks were collected from cattle. The species concerned (three genera and four species of ticks: Haemaphysalis vietnamensis, Haemaphysalis longicornis, Ixodes sinensis and Boophilus microplus) were identified by microscopy and their numbers recorded every month. Because H. longicornis is a three-host tick, with the larvae, nymphs and adults all infesting cows and seasonal fluctuations in nymphs and adults correlating with theileriosis outbreaks, this tick species was a potential vector of T. sergenti. Epidemiological studies indicated that this disease usually occurred in summer or autumn. The disease attacked calves from May to August reaching a peak in June or July (as happened from 1992-1994). In cattle, the disease occurred from June to September with a peak in September. The following pathological lesions were observed: haemorrhages were seen in the subcutaneous tissues and serosa, in the intestinal mucosa and the mesenteric lymph nodes. The surface of the abomasum looked like a piece of red cloth. The liver was enlarged, its edge blunt and curled over. The gall bladder was filled with tawny coloured bile. The spleen was enlarged and dark brown. The heart was enlarged with numerous haemorrhagic foci in the auricle. The blood clotted incompletely. Sick cattle were treated with Berenil and imidocarb and the death rate was reduced from 26% to 5.9%.

Animals↗

Western blot analysis of tick antigens from a Rhipicephalus sanguineus unfed larval extract and identification of antigenic sites in tick sections using immunohistochemistry. A comparative study between resistant and susceptible host species.

Most parasite-host relationships are characterized by the development of resistance by the host, thus limiting the number of parasites. However, some cases are very unusual. In the relationship of the domestic dog with the brown dog-tick Rhipicephalus sanguineus this does not occur, whereas guinea pigs develop efficient resistance. Sera from domestic dogs, crab-eating foxes and guinea pigs collected before and after infestation with R. sanguineus ticks, and after immunization with a whole tick adult or larval homogenate, were used in Western blot analysis to compare and identify potential important antigens from a tick larval homogenate. The same sera were tested in an indirect immunohistochemistry assay in an attempt to compare relevant antigenic sites on histological tick sections. The immunoblotting displayed antigens recognized only by the guinea pigs, as well as several shared antigens between host species, depending on the king of immunization. Immunohistochemistry revealed probable antigenic sites on the cells and tissues of ticks, which varied depending on the kind of immunization (infestation or vaccination) and the animal species involved.

Animals↗

Control of ticks resistant to immunization with Bm86 in cattle vaccinated with the recombinant antigen Bm95 isolated from the cattle tick, Boophilus microplus.

The recombinant Bm86-containing vaccine Gavac(TM) against the cattle tick Boophilus microplus has proved its efficacy in a number of experiments, especially when combined with acaricides in an integrated manner. However, tick isolates such as the Argentinean strain A, show low susceptibility to this vaccine. In this paper we report on the isolation of the Bm95 gene from the B. microplus strain A, which was cloned and expressed in the yeast Pichia pastoris producing a glycosylated and particulated recombinant protein. This new antigen was effective against different tick strains in a pen trial, including the B. microplus strain A, resistant to vaccination with Bm86. A Bm95-based vaccine was used to protect cattle against tick infestations under production conditions, lowering the number of ticks on vaccinated animals and, therefore, reducing the frequency of acaricide treatments. The Bm95 antigen from strain A was able to protect against infestations with Bm86-sensitive and Bm86-resistant tick strains, thus suggesting that Bm95 could be a more universal antigen to protect cattle against infestations by B. microplus strains from different geographical areas.

Animals↗

Integrated control of ticks and tick-borne diseases of cattle in Africa.

The problems caused by tick and tick-borne diseases for livestock particularly cattle on the African continent are described and discussed. The control of ticks and tick-borne diseases must receive high priority in Africa with regard to both research and control application because of their widespread distribution in areas of high livestock potential and productivity. The conventional methods of tick and tick-borne disease control are discussed and are found to be inadequate in the conditions prevailing in Africa. Methods of integrated control are suggested and discussed in light of recent development in control methods and those still under development. Any one of these methods may not be adequate to control the problem on its own but when several of the methods are combined an economic and robust integrated control is likely to result. Encouragement is given to attempt this approach in Africa to solve what must be the largest animal health problem of livestock remaining in the world.

Anaplasmosis↗

Cluster of tick-borne infections at Fort Chaffee, Arkansas: Rickettsiae and Borrelia burgdorferi in ixodid ticks.

Human intrusion into pristine habitats increases the likelihood of acquiring infectious agents from potentially infective ticks. As part of a larger human serological investigation into tick-borne illnesses, 3,000 ixodid ticks were collected during May, August, and November 1990 at Fort Chaffee, Arkansas. Ticks were examined to determine whether they harbor rickettsiae, ehrlichiae, and Borrelia burgdorferi, and to assess relationship to human exposure to tick-borne infections at Fort Chaffee, Ark. The overall tick infection rates with SFG rickettsiae, B. burgdorferi, and ehrlichiae were 4.8, 0.1, and 0.3%, respectively.

Animals↗

Amblyomma testudinarium tick bite: one case of engorged adult and a case of extraordinary number of larval tick infestation.

This paper reports two recent cases of tick bite due to Amblyomma testudinarium. The first case was an 86-year-old farmer infested with a fully engorged adult tick attached on his inguinal region. The second case was a 57-year-old male infested with an extraordinarily large number of larval ticks (> 100 larvae). The ticks were identified as A. testudinarium based on morphological characteristics. To our knowledge, the latter case is the eleventh case of larval tick bites among all tick species and the fourth case with larval A. testudinarium in Japan.

Aged↗

Differences in maturation of tick-borne encephalitis virus in mammalian and tick cell line.

OBJECTIVE: The maturation process of tick-borne encephalitis virus (TBEV) in the tick RA-257 and porcine PS cells was studied by transmission electron microscopy and the E and NS1 proteins were localized in the infected cells. METHODS: The porcine PS and tick RA-257 cell lines were infected with TBEV and examined at different time points post infection under an electron microscope. The E and NS1 proteins were localized with monoclonal antibodies on ultrathin cryosections. RESULTS: The first virus particles and virus-induced vesicles appeared inside hypertrophied and dilated rough endoplasmic reticulum (RER) cisternae in PS cells 15 h p.i. In the course of progressing maturation, the virus particles came up inside the Golgi apparatus and then probably left the cell by the exocytic pathway. Free nucleocapsids did not appear. The observed pattern corresponded to a trans-type maturation. The maximum of the infected PS cell survival was about 50 h p.i. Immunolocalization of some viral proteins (the envelope protein E and the nonstructural protein NS1) revealed the proteins in the cytosol and on the membrane of hypertrophied RER cisternae. On the other hand, the maturation process exhibited different features in the case of the tick RA-257 cells. The nucleocapsids appeared in the cytosol 24 h p.i. and enveloped viral particles were observed in the lumen of vacuoles. Infection of RA-257 cells caused only minor ultrastructural changes and resulted in persistent infection. Immunolocalization of viral proteins in the tick cell line also differed. Proteins E and NS1 were localized in the cytosol and on the vacuolar and plasma membranes. CONCLUSION: The TBEV maturation pathway in the mammalian host cell line differs from the pathway that the virus undergoes in the tick vector cell line.

Animals↗

Importance of ticks and tick-borne diseases of domestic ruminants in Southern Europe.

In Southern Europe, as elsewhere, the importance of ticks and tick-borne diseases (TBD) depends on the species of tick vectors present and also varies to a great extent according to the genetic susceptibility of the livestock involved, as well as other factors such as management and vegetation. There are large gaps in our knowledge of the tick species in the various countries, and of the prevalence and economic importance of the diseases they transmit. There is a great need for tick surveys as well as surveys of tick-borne infections. It is essential to distinguish between infection and disease. A list is given of TBD and their vectors known or to be expected to occur in domestic ruminants in Southern Europe. It is hoped that the exchange of knowledge during this workshop will indicate the importance of these parasites in the various countries involved.

Animals↗

Parasites of domestic and wild animals in South Africa. XL. Ticks on dogs belonging to people in rural communities and carnivore ticks on the vegetation.

Ixodid ticks were collected at weekly intervals from February 1999 to April 2000 from domestic dogs belonging to people in rural communities in north-eastern KwaZulu-Natal. Seven ticks were identified to species level and adult Haemaphysalis leachi followed by immature Amblyomma hebraeum and adult Rhipicephalus simus were the most prevalent. Infestation with adult H. leachi was most intense during February and March 1999 and from January to April 2000, and least intense during October 1999. Infestation with adult R. simus was most intense during February and March 1999 and least during July and September of the same year. For 12 consecutive years, from August 1988 to July 2000, free-living ticks were collected at monthly intervals from the vegetation of two landscape zones in the Kruger National Park. Most adult H. leachi were collected in the one zone from March to June, and no ticks were collected in October, whereas, with the exception of March, most were collected in the other zone during the period January to June, and only one tick was collected in October. Most adult R. simus were collected during February and March at both localities and only a single tick was collected in September. Adult Rhipicephalus turanicus was present only in the one zone and was collected only from January to June.

Animals↗

[Changes in the bactericidal activity of the cementing fraction in ixodid tick saliva under the influence of the presence of the tick-borne encephalitis virus].

It is demonstrated that the abundance of bactericidal compounds in the salivary cement plug of ixodid ticks is changing under the influence of virulent tick-borne encephalitis virus (TBEV) reproduction in their bodies. TBEV infected Ixodes persulcatus cement plug-forming lytic zone size enlarged in comparison with the naive tick one whereas Micrococcus lysodeikticus lytic zones induced by the cement plug of TBEV infected Amblyomma hebraeum nymphs or Rhipicephalus appendiculatus females reduced compared with produced by noninfected specimens. It is supposed that increasing of the lysozyme that production by the main TBEV vectors (Ixodinae ticks) infected salivary gland cells in comparison with suppression of the bactericidal qualities of saliva of the ticks which are not TBEV vectors in the nature (Amblyomminae ticks) is one of markers of the specificity of I. persulcatus-TBE virus interface.

Animals↗

Anti-tick grasses as the basis for developing practical tropical tick control packages.

Of six grass species analysed, Melinis minutiflora (molasses grass) showed the highest anti-tick deterrent properties while Andropogon gayanus (Gamba grass) exhibited the ability to maintain a defined, constantly low, initial host tick infection property and lengthy but low to moderate field tick population. It was concluded that Melinis minutiflora is a species which would best be used in a tick control package within a marginal tick zone while Andropogon gayanus has the advantage within an endemic tick zone.

Animals↗