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PubMed2004

[Molecular cloning of two Rhipicephalus haemaphysaloides haemaphysaloides cathepsin L-like cysteine proteinase gene].

Ticks are obligate ectoparasites and vectors of arboviruses, vickettsiate, spirochetes and parasitil protozoa of humans and domestic animals. Immunological protection of mammalian hosts against tick infestation has been proposed as the most sustainable alternative tick control method to the current use of acaricides. The success of this method is dependent on the identification of key molecules for use as tick vaccine antigens. Proteolytic enzymes are involved in a wide range of cellular processes, thus they can be considered as good target antigens for a tick vaccine. In the present study, we used rapid amplification of cDNA ends protocol and primers that were designed based on the consensus amino acid motifs flanking present in all papain-like cysteine proteinases, to amplify, sequence and characterize two Rhipicephalus haemaphysaloides haemaphysaloides cathepsin L-like cysteine proteinases, named as cysA and cysB. The full length of cysA is 1168bp, encoding a 332 amino acid residue polypeptide with 36.33kD predicted molecular mass; the full length of cysB is 1153bp, encoding a 335 amino acid residue polypeptide with 37.56kD predicted molecular mass. The consensus amino acid motifs flanking presence in both deduced amino acid sequences. And both genes show high sequence homology to other tick cathepsin L-like cysteine proteinase, so they were identified as members of the cysteine proteinase gene family. Expression analysis by RT-PCR revealed that cysA and cysB were expressed differently in different periods of tick development.

Amino Acid Sequence↗
PubMed2006

An antivector vaccine protects against a lethal vector-borne pathogen.

Vaccines that target blood-feeding disease vectors, such as mosquitoes and ticks, have the potential to protect against the many diseases caused by vector-borne pathogens. We tested the ability of an anti-tick vaccine derived from a tick cement protein (64TRP) of Rhipicephalus appendiculatus to protect mice against tick-borne encephalitis virus (TBEV) transmitted by infected Ixodes ricinus ticks. The vaccine has a "dual action" in immunized animals: when infested with ticks, the inflammatory and immune responses first disrupt the skin feeding site, resulting in impaired blood feeding, and then specific anti-64TRP antibodies cross-react with midgut antigenic epitopes, causing rupture of the tick midgut and death of engorged ticks. Three parameters were measured: "transmission," number of uninfected nymphal ticks that became infected when cofeeding with an infected adult female tick; "support," number of mice supporting virus transmission from the infected tick to cofeeding uninfected nymphs; and "survival," number of mice that survived infection by tick bite and subsequent challenge by intraperitoneal inoculation of a lethal dose of TBEV. We show that one dose of the 64TRP vaccine protects mice against lethal challenge by infected ticks; control animals developed a fatal viral encephalitis. The protective effect of the 64TRP vaccine was comparable to that of a single dose of a commercial TBEV vaccine, while the transmission-blocking effect of 64TRP was better than that of the antiviral vaccine in reducing the number of animals supporting virus transmission. By contrast, the commercial antitick vaccine (TickGARD) that targets only the tick's midgut showed transmission-blocking activity but was not protective. The 64TRP vaccine demonstrates the potential to control vector-borne disease by interfering with pathogen transmission, apparently by mediating a local cutaneous inflammatory immune response at the tick-feeding site.

Amino Acid Sequence↗
PubMed1980

Demonstration of Anaplasma marginale Theiler in Dermacentor variabilis (Say) by ferritin-conjugated antibody technique.

Cattle were inoculated with a Virginia isolate of Anaplasma marginale Theiler and served as an infective source for laboratory-reared Dermacentor variabilis (Say) nymphs. Transstadial transmission of A marginale was demonstrated by feeding the newly molted adult ticks on susceptible cattle and by inoculation of gut homogenates collected from adult ticks at postattachment day 6. A similar gut homogenate from the same group of ticks was caused to react with A marginale-bovine antisera that had been conjugated with ferritin. The homogenate contained organisms similar to those observed in the gut of adult feeding ticks that were infected as nymphs and the labeling of the outer membrane confirmed that organisms observed were A marginale. A gut homogenate prepared from control ticks did not cause infection when inoculated into a susceptible cow and no organisms were demonstrated.

Anaplasma↗
PubMed1992

Infectivity of entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) to female ticks of Boophilus annulatus (Arachnida: Ixodidae).

Exposing Boophilus annulatus (Say) to different concentrations of Steinernema carpocapsae (Filipjev) infective juveniles in petri dishes (50-10,000 nematodes per dish) resulted in high mortality (greater than 90%) at nematode concentrations as low as 500 nematodes per dish within 8 d. At a concentration of 10,000 nematodes per dish, 100% of the ticks died within 2 d after infestation. After exposure to 500 nematodes per dish, complete mortality was achieved with the Heterorhabditis bacteriophora strain 'HP88' within 4 d. During the same period, only 15 and 40% mortality were recorded with the 'Mexican' and 'All' strains of S. carpocapsae, respectively. In a lethal dose analysis, S. carpocapsae strain 'DT' was the most infective strain with the lowest LD50 and LD90 values (15 and 165 infective juveniles per tick, respectively). The 'All' strain of S. carpocapsae was the least infective of the four strains tested, with LD50 and LD90 values of 372 and 9,251 infective juveniles/tick, respectively. Optimal temperature for tick control by the nematodes was between 22 and 26 degrees C. Mortality rate was reduced at 18 and 30 degrees C. The susceptibility of fully engorged ticks was not influenced by the weight of the replete females. Nematode infection did not have an adverse effect on egg laying by surviving ticks.

Animals↗
PubMed2001

The distribution of ixodid ticks (Acari: Ixodidae) in central Ethiopia.

Ixodid ticks were collected from domestic animals, mainly cattle, in 11 administrative zones covering 84 districts in central Ethiopia over a period of 2 years (July 1996 to June 1998). Nineteen tick species were identified. Four of these belonged to the genus Amblyomma, one to Boophilus, two to Haemaphysalis, three to Hyalomma and nine to Rhipicephalus. Amblyomma variegatum and Rhipicephalus evertsi evertsi were present in all 11 administrative zones and, with the exception of Afar, Boophilus decoloratus was present in nearly every district in which collections were made. These three species constituted more than 50% of all ticks collected. Amblyomma cohaerens and Rhipicephalus bergeoni were common in the west of the survey region and Rhipicephalus pulchellus in the east. Except for B. decoloratus, of which more females than males were collected, the numbers of male ticks recovered were equal to or exceeded those of females. Mortality in crossbred dairy cattle caused by heartwater (Cowdria ruminantium infection) was reported during the survey period. An integrated approach to tick control is suggested.

Animals↗
PubMed1999

Molecular cloning of two Haemaphysalis longicornis cathepsin L-like cysteine proteinase genes.

Immunological protection of mammalian hosts against tick infestation has been proposed as the most sustainable alternative tick control method to the current use of acaricides which has several limitations. The success of this method is dependent on the identification of key molecules for use as tick vaccine antigens. Proteolytic enzymes are involved in a wide range of cellular processes in eukaryotes such as development regulation and nutrition, thus they can be considered as good target antigens for a tick vaccine. In the present study we used primers designed based on the consensus amino acid motifs flanking the conserved active sites C25 and N175 present in all papain-like cysteine proteinases to amplify by polymerase chain reaction, sequence and characterize two Haemaphysalis longicornis tick cysteine proteinase genes. Based on the nucleotide and deduced amino acid sequences, both genes were identified as members of the cysteine proteinase gene family by presence in their sequences of consensus motifs flanking the conserved active sites C25, H150 and N175 that are present in all papain-like cysteine proteinases. Both genes are about 1.2 kb in size and show high sequence homology predominantly to invertebrate cathepsin L-like cysteine proteinases.

Amino Acid Sequence↗
PubMed2005

A serine protease inhibitor (serpin) from Haemaphysalis longicornis as an anti-tick vaccine.

The application of anti-tick vaccine has been shown to be the most promising alternative tick control strategy compared to the current use of acaricides that suffer from a number of limitations. The success of this strategy is dependent on the cloning, and characterization of tick molecules involved in the mediation of tick central physiological roles. Rapid amplification of the cDNA ends (RACE) and primers designed based on a conserved serpin amino acid motif (NAVYFKG) were used to clone a cDNA with high similarity in the reactive center loop (RCL) to representative serpin, heparin cofactor II. We have named this novel gene as Haemaphysalis longicornis serpin-2 (HLS2). RT-PCR analysis showed that HLS2 mRNA transcripts are not expressed in salivary glands but in hemolymph by feeding ticks. HLS2 was not introduced into the bite site as measured by Western blot analysis. The activated partial thromboplastin time (APTT) and the thrombin inhibitory assay using recombinant HLS2 (rHLS2) demonstrated prolonged coagulation time and inhibition of thrombin activity. These results suggested that HLS2 is present only in hemolymph of the feeding ticks and the function of HLS2 is homeostasis in tick physiological compartment. Vaccination of rabbits with rHLS2 conferred protective immunity against ticks, resulting in 44.6 and 43.0% mortality in nymphal and adult ticks, respectively. These results show that rHLS2 could be an important candidate as a component of a cocktail anti-tick vaccine.

Amino Acid Sequence↗
PubMed1991

The efficacy of ultrasonic pest controllers for fleas and ticks.

Two ultrasonic pest controllers, a pet-collar unit and a large unit for household use, were tested for their efficacy in repelling fleas and ticks in a choice chamber. Neither unit had any affect on the distribution of fleas or ticks in the choice chamber up to 24 h exposure, and activity of fleas, ticks and cockroaches was unimpaired. The study extends and supports previous findings that ultrasound is ineffective as a means of controlling common pests of households and pets.

Animals↗
PubMed1998

Individual host variations in tick infestations of cattle in a resource-poor community.

Relative resistance levels of cattle against tick infestations in the communal grazing area of Botshabelo in the south-eastern Free State were determined. The objective was to establish whether differences in resistance can be exploited to contribute to tick control methods used by small-scale farmers in resource-poor environments. Ten cows (Bos taurus crosses) between the ages of 18 months and four years were used and tick counts were conducted once a month over a period of 12 months to compare their total tick burdens. Tick burdens of the various animals were compared mutually as well as with the mean tick burden of the group as a whole. Tick numbers varied throughout the year on all individuals but some animals consistently tended to have either higher or lower numbers than the mean of the group. Tick burdens on cattle classified as having a relatively low resistance to tick infestations increased eleven-fold from January to June 1996 compared to a six-fold increase on cattle categorized as belonging to the high resistance group. Twenty-eight percent of the cattle in the total study group carried 50% of the ticks collected (60,079). It is recommended that farmers in the region visually assess B. decoloratus burdens, the most abundant tick species, and sell or cull the most susceptible animals first in their normal program of utilization of the animals. This should eventually result in the direct improvement of the overall tick resistance of their cattle herds.

Analysis of Variance↗
PubMed2006

[Ixodes ricinus. Health risks and approaches in prevention].

Ticks belong to the parasitical mites. There are more than 1000 different species worldwide which do need blood of warm-blooded organisms for their own development. As ticks are well known to be vectors of several pathogens which could cause insidious infectious diseases, they are important in human and veterinarian medicine as well. It is not possible to control ticks in nature. Therefore the first step should be an effective prevention. In case of a bite the tick must be removed as soon as possible. Although Ixodes ricinus is more or less responsible for all health risks caused by ticks in Germany, further species like Rhipicephalus sanguineus, Dermacentor marginatus and Argas reflexus must not be forgotten, as these ticks have appeared more frequently in the recent past and they can be vectors of several diseases as well. This paper describes first and foremost the biological behaviour and development of Ixodes ricinus and the medical importance of this tick in human and veterinarian medicine.

Animals↗
PubMed2005

Purification and antigenicity of two recombinant forms of Boophilus microplus yolk pro-cathepsin expressed in inclusion bodies.

The tick Boophilus microplus is a bovine ectoparasite present in tropical and subtropical areas of the world and the use of vaccines is a promising method for tick control. BYC is an aspartic proteinase found in eggs that is involved in the embryogenesis of B. microplus and was proposed as an important antigen in the development of an anti-tick vaccine. The cDNA of BYC was amplified by PCR and cloned for expression in two forms with and without thioredoxin fusion protein (Trx), coding recombinant proteins named rBYC-Trx and rBYC, respectively. Expression, solubility, and yields of the two forms were analyzed. The recombinant proteins were expressed in inclusion bodies (IBs) and three denaturant agents (N-lauroyl sarcosine, guanidine hydrochloride, and urea) were tested for IBs solubilization. The N-lauroyl sarcosine solubilized 90.4 and 92.4% of rBYC-Trx and rBYC IBs, respectively, and was the most efficient denaturant. Two recombinant forms were affinity-purified by Ni2+-Sepharose under denaturing conditions. After dialysis, the yield of soluble protein was 84.1% for r-BYC-Trx and 5.9% for rBYC. These proteins were immune-reactive against sera from rabbit, mouse, and bovine previously immunized with native BYC, which confirms the antigenicity of the recombinant BYCs expressed in the Escherichia coli system.

Animals↗
PubMed1978

Nairobi sheep disease in Kenya. The isolation of virus from sheep and goats, ticks and possible maintenance hosts.

Nairobi sheep disease was seen principally upon movement of susceptible animals into the enzootic areas. This occurred most frequently for marketing purposes near the main centres of population. Other outbreaks followed local breakdowns in tick control measures. The disease did not occur in epizootic form during the period under consideration. Nairobi sheep disease was isolated from pools of Rhipicephalus appendiculatus but not from many pools of other tick species. No virus was isolated from the blood or tissues of a range of wild ruminants and rodents.

Animals↗
PubMed1986

Ixodid-host immune interaction. Identification and characterization of relevant antigens and tick-induced host immunosuppression.

Ixodid ticks are the most important vectors of pathogens to domestic and wild animals. It is established that cattle and laboratory animal species acquire resistance to tick infestation; acquired resistance has an immunological basis consisting of cell-mediated, antibody-mediated and complement-dependent effector mechanisms. Even though acquired resistance to tick feeding is expressed, host immune competence is possibly impaired during the course of tick feeding. Ixodid-induced transient immunosuppression could possibly facilitate the transmission of vector-borne pathogens and/or enhance tick feeding capabilities in the presence of a host immune response to the hematophagous arthropod. Tick tissue extracts have been used to artificially induce resistance to ixodid feeding, and this has become an area of increasing interest as a possible strategy for tick control. It is essential to have defined antigenic molecules for analysis of host responses to infestation, characterization of immunopathologic processes and for vaccine development. This report focuses on attempts to identify, characterize and isolate tick immunogens. Protein immunoblotting, utilizing sera from animals of different genetic composition and infestation patterns, was used to detect a number of tick polypeptides which are reactive with sera of infested hosts. It is clear that infestation with one ixodid species stimulates antibodies reactive with molecules derived from the sensitizing species and/or tick species in the same genus or different genera. This approach is used to identify molecules that are good candidates for use in immunization studies and for analysis of mechanisms involved in acquisition and expression of resistance to tick feeding.

Animals↗
PubMed1980

The subgenus Persicargas (Ixodoidea: Argasidae: Argas). 33. Effect of gamma radiation on first nymphal instar A. (P.) arboreus and adult fertility.

Gamma radiation doses higher than 1,000 rads are lethal to first nymphal instar Argas (Persicargas) arboreus. Only 50% of first nymphal instars receiving 1,000 rads reach adulthood but those receiving 100-500 rads survive normally. Males resulting from irradiated first nymphal instars are almost normally fertile. Female germinal cells tolerate higher radiation doses received in the first nymphal instar stage than in the adult stage. Females resulting from first nymphal instars receiving does higher than 100 rads are less fertile (egg number and percent hatch) than normal and produce F1 larvae of lower than normal viability. Progenies of females irradiated as nymphs apparently inherit lethal genes, which may be useful, if irradiated at critical stages, in tick control.

Animals↗
PubMed1991

Predation of free-living engorged female Rhipicephalus appendiculatus.

In experiments done over a period of 1 1/2 years using engorged female Rhipicephalus appendiculatus tethered in a grass plot, 42% predation was observed in long grass (40-60 cm), and 36% in short grass (6-10 cm). Deaths due to environmental factors were 4.8% and 6.8% in long and short grass, respectively. Six groups of animals were confirmed to be predators of the ticks, namely: ants, spiders, rodents, birds, lizards and shrews. The implications of these results in making tick population models, and the possibility of using predators in integrated tick-control packages are discussed.

Animals↗
PubMed1993

Temperature- and humidity-dependent longevity of unfed adult Hyalomma truncatum (Acari: Ixodidae).

The survival of unfed adult Hyalomma truncatum Koch held under different regimes of constant temperature (5, 17, 24, and 30 degrees C) and relative humidity (10, 50, and 80%) was monitored during > 1 yr. Longevity of this medically important African tick was shortest at the highest temperature and lowest relative humidity (100% dead at week 25). Conversely, H. truncatum lived longest at lower temperatures and higher relative humidity (< 100% dead at week 64). The combined effects of temperature and humidity, measured as vapor pressure deficit, were strongly related to survival of these ticks. The survival of males and females was similar and was independent of the weight of ticks. These findings have implications for the maintenance and study of laboratory colonies of H. truncatum and for the development of tick control strategies to reduce vectorial capacity.

Animals↗
PubMed1999

Characterization of proteolytic enzymes expressed in the midgut of Haemaphysalis longicornis.

The proteolytic activities present in midguts of both fed and unfed Haemaphysalis longicornis were assessed by using the gelatin-substrate gel electrophoresis and inhibitor sensitivity analyses. Three predominant (116, 48 and 48 kDa) and two weak (55 and 60 kDa) proteinase bands were commonly expressed in both unfed and fed ticks, while a weak 80 kDa band was only present in fed ticks. Consistent with observations on other tick species, proteolytic activity against the gelatin substrate was observed only under acidic conditions. Inhibition studies against the gelatin substrate using a panel of inhibitors showed that the predominant proteolytic enzymes of 40 and 48 kDa molecular mass are cysteine proteinases. These results are discussed in the context of host vaccination as an alternative tick control method to the current use of chemical acaricides.

Animals↗
PubMed2001

Feeding aggregation of the tick Rhipicephalus appendiculatus (Ixodidae): benefits and costs in the contest with host responses.

Gregariousness can be advantageous in interspecific competition while intraspecific competition may favour solitude. We examined feeding behaviour of the ixodid tick, Rhipicephalus appendiculatus, in the context of interspecific (tick-host) and intraspecific (tick-tick) competition. Such competition is mediated through host rejection responses to tick infestation to which ticks respond by secreting immunodulatory saliva. We observed that group feeding adults increased their blood-feeding rate, reducing the time to mating and repletion, compared with individual feeding of paired adults. The benefits of feeding aggregation indicate direct reciprocity between ticks, most likely resulting from the shared activities of their bioactive saliva. However, fast-feeding ticks appeared to impair blood-feeding success of slow-feeding females during group feeding. This may be explained by the faster feeders exacerbating host responses on detachment that are then directed against the slower feeders. As female fecundity is generally proportional to the size of the bloodmeal, there will be a selection pressure to feed gregariously. Greater understanding of the benefits and costs of feeding aggregation may help to improve tick control strategies.

Animals↗