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Isolation of protective antigens from the gut of Boophilus microplus using monoclonal antibodies.

Monoclonal antibodies (mAb) were produced against midgut membrane (GM) antigens of Boophilus microplus. The isotypes of these mAb were established and their specificity characterized using double diffusion and Western blotting. GM antigens solubilized by Triton X-100 were precipitated by mAb QU13, and the precipitate was then injected into cattle to test for the presence of protective antigens. Vaccinated cattle challenged with 10-day-old larval ticks showed evidence of protection with a 62% reduction in eggs produced by ticks from vaccinated cattle compared to tick eggs from control cattle. In a second vaccine-challenge experiment, the dose of precipitate was increased and greater than 99% protection was provided to these vaccinated cattle following challenge (calculated from tick egg weights compared to the control group). The solubilized antigen(s) precipitated by QU13 were subjected to SDS-PAGE separation and the calculated sizes of these molecules were greater than 200,000, 80,000, 74,000, 62,000 57,000 and less than 30,000 MW.

Animals↗

Factors that influence the prevalence of acaricide resistance and tick-borne diseases.

This manuscript provides a summary of the results presented at a symposium organized to accumulate information on factors that influence the prevalence of acaricide resistance and tick-borne diseases. This symposium was part of the 19th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP), held in New Orleans, LA, USA, during August 10-14, 2003. Populations of southern cattle ticks, Boophilus microplus, from Mexico have developed resistance to many classes of acaricide including chlorinated hydrocarbons (DDT), pyrethroids, organophosphates, and formamidines (amitraz). Target site mutations are the most common resistance mechanism observed, but there are examples of metabolic mechanisms. In many pyrethroid resistant strains, a single target site mutation on the Na(+) channel confers very high resistance (resistance ratios: >1000x) to both DDT and all pyrethroid acaricides. Acetylcholine esterase affinity for OPs is changed in resistant tick populations. A second mechanism of OP resistance is linked to cytochrome P450 monooxygenase activity. A PCR-based assay to detect a specific sodium channel gene mutation that is associated with resistance to permethrin has been developed. This assay can be performed on individual ticks at any life stage with results available in a few hours. A number of Mexican strains of B. microplus with varying profiles of pesticide resistance have been genotyped using this test. Additionally, a specific metabolic esterase with permethrin-hydrolyzing activity, CzEst9, has been purified and its gene coding region cloned. This esterase has been associated with high resistance to permethrin in one Mexican tick population. Work is continuing to clone specific acetylcholinesterase (AChE) and carboxylesterase genes that appear to be involved in resistance to organophosphates. Our ultimate goal is the design of a battery of DNA- or ELISA-based assays capable of rapidly genotyping individual ticks to obtain a comprehensive profile of their susceptibility to various pesticides. More outbreaks of clinical bovine babesisois and anaplasmosis have been associated with the presence of synthetic pyrethroid (SP) resistance when compared to OP and amidine resistance. This may be the result of differences in the temporal and geographic patterns of resistance development to the different acaricides. If acaricide resistance develops slowly, herd immunity may not be affected. The use of pesticides for the control of pests of cattle other than ticks can affect the incidence of tick resistance and tick-borne diseases. Simple analytical models of tick- and tsetse-borne diseases suggest that reducing the abundance of ticks, by treating cattle with pyrethroids for example, can have a variety of effects on tick-borne diseases. In the worst-case scenario, the models suggest that treating cattle might not only have no impact on trypanosomosis but could increase the incidence of tick-borne disease. In the best-case, treatment could reduce the incidence of both trypanosomosis and tick-borne diseases Surveys of beef and dairy properties in Queensland for which tick resistance to amitraz was known were intended to provide a clear understanding of the economic and management consequences resistance had on their properties. Farmers continued to use amitraz as the major acaricide for tick control after the diagnosis of resistance, although it was supplemented with moxidectin (dairy farms) or fluazuron, macrocyclic lactones or cypermethrin/chlorfenvinphos.

Acaricides↗

The susceptibility of different species and stages of ticks to entomopathogenic fungi.

Boophilus annulatus, Hyalomma excavatum and Rhipicephalus sanguineus ticks were shown to be susceptible to different entomopathogenic fungi under laboratory conditions. Comparative results of bioassays using five different fungal species showed that some strains of Metarhizium anisopliae are highly pathogenic against various tick stages tested. In contrast to to their activity against insects, fungi also affected tick eggs. All tested tick stages including those feeding on a host were found to be susceptible to these fungi, except for adult H. excalatimn ticks, which were relatively resistant.

Animals↗

Computer simulation of management strategies for American dog ticks (Acari: Ixodidae) and Rocky Mountain spotted fever.

Computer models were developed to simulate the effects of management technologies on populations of the American dog tick, Dermacentor variabilis (Say), principal vector of Rocky Mountain spotted fever (RMSF) in eastern North America. The technologies modeled were area-wide acaricide application, acaricide-food-baited tubes for self-treatment by small mammals, dipping of dogs in acaricides, acaricide-impregnated plastic dog collars, reduction of small mammal host populations (host management), and removal of vegetation that protects free-living tick stages (vegetative management). Submodels for each of these technologies were incorporated into a model (ADTSIM) for the population dynamics of the tick and RMSF transmission. Comparisons of simulated and observed data were used to verify reasonable accuracy of the submodels. Repetitive simulations were made to identify levels and timing of each control method (alone or combined) required to reduce tick populations below a RMSF transmission threshold of 252 unfed adults/ha. Eight to 30 acaricide applications, depending on acaricide and percentage of population treated, were needed during a 10-yr period to reduce densities of ticks below the threshold. The baited-tube method, host management, and vegetative management (depending on level and frequency of treatment) also were capable of reducing tick density below the threshold. However, acaricide-impregnated plastic dog collars did not reduce tick density below the threshold unless at least 50% of the hosts of adult ticks were domestic dogs. Integrated strategies were developed for management of ticks and RMSF in six selected states. These strategies reduced numbers of human cases of RMSF 90% or more by year 20 by maintaining tick densities between 100 and 252 unfed adults/ha.

Animals↗

[In vitro patogenicity of the fungi Metarhizium anisopliae (Metschnikoff, 1879) Sorokin, 1883, on the tick Boophilus microplus (Canestrini,1887)].

The cattle tick Boophilus microplus is a serious concern to the Brazilian cattle industry. It causes decreased meat and milk production, low alimentary conversion, damage to leather and transmission of pathogens. The use of entomopathogenic fungi as biological control agents for ticks has shown promising results. This study tests the in vitro effects of two Metarhizium anisopliae isolates (E9 and 319) towards three stages of B. microplus: eggs, larvae and engorged females. The bioassays were composed of five treatment groups (concentrations: 10(5), 10(6), 10(7), 10(8) conidia/ml and the control) each one with ten repetitions. The stages were treated by immersion for three minutes. After the treatments, the females, eggs and larvae were incubated at 27 degrees C +/- 1 and RH >or= 80%. The main studied parameters were: percentage of larvae eclosion, percentage of larvae mortality and the indexes of nutritional and reproductive efficiency. In vitro tests of the fungal isolates showed effective control of the three stages of B. microplus, suggesting high potential for their use as a microbial control agent of the B. microplus tick.

Animals↗

New member of the protein disulfide isomerase (PDI) family identified in Amblyomma variegatum tick.

Ticks belonging to arthropoda are blood feeding, geographically widespread ectoparasites of mammals, reptiles and birds. Their saliva contains active substances that protect them from host immune attack and allow for transmission of various pathogens during the feeding process. Characterization of tick saliva components can therefore contribute to the development of effective methods for the control of tick-borne diseases. Here we describe the identification and basic characterization of a gene encoding a 55kDa protein found in the salivary glands (SG) of Amblyomma variegatum tick. Based on the primary structure and homology to the family of protein disulfide isomerases (PDI; EC 5.3.4.1) the gene was named AvPDI. The 1461nt long AvPDI open reading frame codes for a 487 amino acid protein. In vitro expressed AvPDI was exclusively localized in the endoplasmic reticulum. RT-PCR and Western blot analysis revealed that AvPDI expression is not restricted to the SG of the tick. More detailed analysis on tissue slides from SG detected an AvPDI specific signal in granular cells of the acini type II and III. Finally, reductase activity of AvPDI was confirmed in an insulin assay. The structural and functional characteristics suggest that AvPDI is another member of the PDI protein family and represents the first more closely characterized PDI in the ticks.

Amino Acid Sequence↗

Cloning, expression and partial characterization of a Haemaphysalis longicornis and a Rhipicephalus appendiculatus glutathione S-transferase.

The ticks Haemaphysalis longicornis and Rhipicephalus appendiculatus are important parasites worldwide. The current method for control of cattle ticks involves the use of chemicals. Nevertheless, parasite resistance is an ever increasing global problem. Glutathione S-transferases (GSTs) play a central role in detoxication of xenobiotic and endogenous compounds. Several authors have noted that an increase in GST activity is associated with resistance to insecticides and acaricides. In the present study, we report the cloning and expression of GST cDNAs from H. longicornis and R. appendiculatus. In addition, we determine the effect of three acaricides (ethion, deltamethrin and diazinon) on the enzymatic activity of rGSTs.

Amino Acid Sequence↗

The effect of veld-burning on the seasonal abundance of free-living ixodid ticks as determined by drag-sampling.

A supervised veld-burn in the Sclerocarya caffra/Acacia nigrescens Savanna landscape zone in the south-eastern region of the Kruger National Park was carried out during September 1988. The effect of the fire on the free-living tick population was determined by comparing the numbers of ticks collected by monthly drag-sampling in the burnt zone with those collected in an adjacent unburnt zone over a 2-year period. A total of 13 ixodid tick species were involved. Tick numbers were reduced after the burn but rose again after varying periods of time. The length of these periods depended upon a number of variables. These included tick species, patterns of seasonal abundance, and host preferences. The original reduction in numbers seemed to result in subsequent cyclical population fluctuations and in some instances overcompensation was noted. Veld-burning as a control technique may be effective with tenuously adapted tick species or reduced populations and may be enhanced by the exclusion of major hosts for a critical period after the fire.

Animals↗

Studies on Theileriidae (Sporozoa) in Tanzania. VI. Second field trial on immunization against cattle theileriosis.

Nine cattle, immunized by an infection and treatment method with 3 strains of Theileria parva (Muguga, Kiambu 5 and Serengeti transformed) and nine controls, were exposed to natural tick infestation in Tanzania. All controls contracted fatal East Coast Fever within 2 months of exposure. All immunized animals survived the period of exposure (over 2 months), but later one died of ECF, one of heart-water, one of an undetermined cause (not a theileriosis), and one disappeared from the herd. The 5 remaining animals survived for over 3 months after exposure ended. It is thought that this method of immunization is of value in protecting valuable animals at risk. An attempt to immunize against pathogenic Theilereia mutans, by injecting blood containing an apathogenic strain of this parasite, gave inconclusive results as no pathogenic strains were encountered during the trial.

Animals↗

Equine piroplasmosis an update on diagnosis, treatment and prevention.

Two haemoprotozoan parasites, Babesia caballi and Babesia equi, can cause equine piroplasmosis. Due to the presence of potential tick vectors in areas so far unaffected by equine babesias, import and export regulations often require the serum testing of animals for evidence of infection. Although the complement fixation test (CFT) has been recommended for detecting the presence of antibodies to Babesia spp., it has been demonstrated to have several disadvantages, including false-positive results and low sensitivity for detecting latent infections. An enzyme-linked immunosorbent assay (ELISA) may be an alternative for increased and sensitive detection of acute and latent babesial infections, but its development to date has been hindered by a limited antigen supply and poor specificity. In vitro cultivation of both parasite species and the identification of parasite proteins for diagnostic use has facilitated the development of a highly sensitive and specific ELISA. For the direct detection of the parasites, DNA probes are now available. Several drugs are available for the treatment of equine piroplasmosis. For instance, diminazene diaceturate is effective in the chemosterilization of B. caballi and in the elimination of clinical signs in B. equi infections. Antitheilericidal drugs such as buparvaquone have been demonstrated to be effective in combatting disease due to B. equi and may--in combination with imidocarb--also eliminate the parasite. The control of equine piroplasmosis must include effective tick control, seromonitoring of animals and the application of chemotherapy.

Animals↗

Moxidectin pour-on for control of natural populations of the cattle tick Boophilus microplus (Acarina: Ixodidae).

Fifty Bos taurus x Bos indicus heifers naturally infested with Boophilus microplus ticks were divided into two groups of 25 heifers each. Individuals of one group were treated with moxidectin 0.5% pour-on at a dosage of 500 microg of moxidectin/kg body weight and heifers from the other group remained as untreated controls. An efficacy higher than 95% was found on days 7-21 after treatment by using female ticks 4.5-8.0 mm long as the main infestation parameter. A lower, but significant efficacy (p < 0.05) was also found on days 1 (32.3% efficacy) and 27 (73.4% efficacy) post-treatment. Significantly (p < 0.05) lower numbers of immature ticks were also observed on heifers of the treated group from days 7 through 27 after treatment. A lower engorgement weight of female ticks from treated heifers was found on days 1 and 21 after treatment. Treatment also affected reproductive performance (oviposition, egg hatch and number of eggs laid) of female ticks collected on Day 1.

Administration, Topical↗

[The use of smells of plant origin as indicators of the extensiveness and intensity of ixodid tick infection with the tick-borne encephalitis virus].

It has been shown that, using a simple olfactometer, through which moist air and smell of vegetative origin penetrate into the centre via different ends of the tube, ticks can be divided into two groups, those infected and not infected with tick-borne encephalitis virus. The method yielding a 100% division, is based on opposite changes in the behaviour of virus infected ticks: the attracting smells become repellent and vice versa. Normal reaction to smell measured by the length of the distance covered by an individual tick towards or from the smell source in the olfactometer tube varies in various groups of control and experimental ticks, but correlates with virus titre of ticks. It is greater in ticks with a higher level of virus reproducibility. The reaction to smell in infected ticks makes it possible to predict the intensity of their infestation.

Animals↗

Larval membrane antigens protect Hereford cattle against infestation with Boophilus microplus.

Hereford cattle (Bos taurus) were immunized with antigens solubilized with Triton X-100 from larval membranes of the cattle tick (Boophilus microplus). Based on tick egg production compared to control cattle, vaccinated cattle were protected (78%) against challenge with 2 x 20,000 tick larvae. The soluble Triton X-100 extract of tick larval membranes was further purified by immunoaffinity chromatography, using immunoglobulin ligands (IgG1 and IgG2) from three immune steers, previously vaccinated with membrane antigens from the midgut of partly engorged adult female ticks. Cattle vaccinated with these purified antigens were protected in two separate experiments (80 and 89% respectively), against challenge with 2 x 20,000 larval ticks compared to control cattle. Whole larval membranes used as vaccines in cattle reduced the amount of eggs produced from ticks by 47% compared to control cattle, but this difference was not significant.

Animals↗

Acaricide resistance--back to basics.

Resistance like life is poorly understood and like death is inevitable (but will take varying times to develop). Resistance develops rapidly where the gene frequency for resistance is high and there is increased selection pressure. Resistance should be seen in perspective and can be managed. For example resistance is most frequent in single-host ticks which transmit less serious diseases. Indeed where products are also used to control tsetse, it may be more important to have live cattle with the risk of tick resistance, (which may in any case take years to develop) than severe losses through trypanosomiasis. Good management can prolong the active life of compounds and the following practical considerations should be adopted:- Field training to ensure the correct usage of products. Ensure ticks are not transported with animals. Establish tick resistance testing centres.

Animals↗

Prevention of tick-borne diseases.

Ticks are a part of the landscape where humans live, work, and play. Because ticks carry a wide range of organisms that potentially can cause disease in humans, many studies have focused on ways to reduce risk of these diseases. Ticks have biologically complex interactions with microorganisms and with their vertebrate hosts, on whom they depend for blood meals and survival. To consider ways to reduce the burden of tick-borne diseases in humans, it is necessary to understand the biology and ecology of ticks and their interface with humans. In many areas, changes in land use, reforestation, and patterns of human settlements have led to more abundant tick populations, increasing rates of infections in ticks, and increasing contact with human populations. Warmer winter temperatures in temperate regions may extend the transmission season for some ticks and pathogens. Although much of the discussion in this article has focused on I. scapularis and the Lyme disease spirochete (because they have been studied extensively), other tick-pathogen pairs may differ in risk factors for infection and transmission dynamics. Interventions studied to reduce the burden of tick-borne diseases include changing the environment, controlling vertebrate hosts, killing ticks, altering the behavior of humans, treating tick bites, and trying to protect humans through immunologic means (vaccine). All of these approaches have limitations and drawbacks. From a public health perspective, a plan that employs multiple strategies may be most effective. This article has reviewed what is known about preventive interventions, including the vaccine.

Animals↗

Resistance to tick-borne Francisella tularensis by tick-sensitized rabbits: allergic klendusity.

Mammals become hypersensitive to ticks that feed upon them. That hypersensitivity was thought responsible for an observation that a large number of Francisella tularensis-infected Dermacentor variabilis failed to infect a rabbit previously exposed to ticks of that species. In a series of tests of that hypothesis, rabbits sensitized to ticks were often significantly more resistant than control animals to tick-borne tularemia. The conditions that determine the klendusity are thought to be variable and complex but the phenomenon must be of importance in the epidemiology of some arthropod-borne agents.

Animals↗