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cDNA cloning, characterization and vaccine effect analysis of Haemaphysalis longicornis tick saliva proteins.

Immunological control of ticks is currently the only sustainable and practical alternative method to the current use of acaricides which has serious limitations. The success of this method is dependent upon identification and cloning of potential tick vaccine antigens. We used a combination of immuno-screening of an adult tick cDNA library as well as the 3 and 5 rapid amplification of cDNA ends to clone two cDNAs, encoding tick saliva proteins from Haemaphysalis longicornis. The two cDNAs herein named HL 34 and 35 are 1000 bp each and encode polypeptides with 292 and 321 amino acid residues respectively. Northern blotting analysis of total RNA from ticks at different feeding stages revealed that expression of both HL 34 and HL35 mRNAs is induced during the slow feeding phase. We speculate that the functions of both genes are closely associated with blood feeding. Expression analysis by RT-PCR showed that both genes are expressed in other tick organs in addition to salivary glands. Recombinant HL 34 was successfully expressed in Escherichia coli and its suitability as a tick vaccine antigen was analyzed in rabbits. We propose that rHL34 could be a useful component of a cocktail tick vaccine.

Amino Acid Sequence↗

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↗

Identification of novel tick salivary gland proteins for vaccine development.

Methods currently used to control Ixodes scapularis ticks rely principally on acaricidal applications which suffer from a number of limitations. Recently, host vaccination against ticks has been shown to be a promising alternative tick control method. In tick salivary glands, numerous genes are induced during the feeding process. Many of these newly expressed proteins are secreted in tick saliva and may play a role in modulating host immune responses and pathogen transmission. We have performed suppression subtraction hybridization to identify unique I. scapularis salary gland proteins specifically expressed during engorgement. We have cloned and sequenced ten unique salivary gland-associated cDNAs that are up-regulated during feeding. The protein products of these genes represent potential vaccine candidates for use in the control of ticks and to prevent transmission of tick-borne diseases.

Amino Acid Sequence↗

Seasonal abundance of ticks associated with indigenous goats on a northern Transvaal farm.

The 3 most abundant tick species on indigenous goats on a northern Transvaal farm were found to be Rhipicephalus evertsi evertsi, Rhipicephalus appendiculatus and Amblyomma hebraeum. Three other tick species were present in small numbers. The economical and ecological importance of the ticks found on the goats, mainly in rural areas where chemical control of ticks is practically non-existant, is discussed. The high number of goats in the study area and the shift towards alternative methods of tick control, such as the use of resistant hosts, are important factors in the livestock industry of southern Africa.

Animals↗

An in vitro feeding assay to test acaricides for control of hard ticks.

Animal husbandry could not be practised over large areas of the planet without acaricides. The prevention of tick bite and the transmission of diseases requires the use of pesticides, but this contributes to the development of tick resistance against acaricides. This drives the quest for new molecules that target physiological processes crucial to tick survival. In vivo trials involve multiple repetitions because of inherent variations between host animals, requiring large amounts of test products and ticks. An in vitro alternative should permit the testing of the ability of a product to restrict attachment and feeding by ticks at precise doses. In this paper an in vitro feeding system is described where the European tick Ixodes ricinus L. feeds on blood through a cellulose rayon-reinforced silicone membrane. The membrane Shore hardness is modified to imitate the elastic retraction forces of skin that ensure the closing of tick penetration sites on the membrane to prevent bleeding. Tick attachment (75-100%) is achieved by adding chemical and mechanical stimuli to the membrane. Survival curves for different doses of fipronil and ivermectin tested with the method showed highly reproducible acaricide effects within 5-7 days. Significant effects are recorded down to ppb levels in blood. Standardised tests can be made with blood from the same donor animal or culture medium under the membrane.

Animals↗

Pheromone/acaricide mixtures in the control of the tick Amblyomma hebraeum: effects of acaricides on attraction and attachment.

Unfed adults and nymphs of the bont tick Amblyomma hebraeum Koch are attracted to hosts on which fed males, emitting an aggregation-attachment pheromone (AAP), are attached. Pheromone/acaricide mixtures have the potential to selectively attract and kill these ticks. We have investigated the effects of three acaricides, amitraz (an amidine), flumethrin (a synthetic pyrethroid) and chlorfenvinphos (an organophosphate), combined with AAP, on the attraction and attachment of the unfed adults. Attraction, measured in field assays involving AAP and CO2, was not inhibited by any of the acaricides. Attachment was measured over 24 h on the ears of rabbits which had been treated with AAP and mixtures of AAP and the acaricides. None of the acaricides inhibited initial attachment. Flumethrin caused rapid and high mortality in attached and unattached ticks. Chlorfenvinphos caused little mortality in the first 24 h on the ears of the rabbits, but 75% of the exposed ticks died over the next 14 days while held in an incubator. Amitraz caused the ticks to detach after an initial period of attachment; there was little mortality in the detached ticks over the next two weeks and they could later be induced to re-attach to other rabbits. Flumethrin was considered to be the compound of choice for use in pheromone/acaricide mixtures.

Animals↗

Ticks feeding on humans: a review of records on human-biting Ixodoidea with special reference to pathogen transmission.

In this article, literature records of argasid and ixodid ticks feeding on humans worldwide are provided in view of increased awareness of risks associated with tick bites. Ticks can cause paralyses, toxicoses, allergic reactions and are vectors of a broad range of viral, rickettsial, bacterial and protozoan pathogens. Approximately 12 argasid species (Argas and Ornithodos) are frequently found attached to humans who intrude into tick-infested caves and burrows. Over 20 ixodid tick species are often found on humans exposed to infested vegetation: four of these are Amblyomma species, 7 Dermacentor spp., 3 Haemaphysalis spp., 2 Hyalomma spp. and 6 Ixodes species. Personal protection methods, such as repellents and acaricide-impregnated clothing are advised to minimize contact with infected ticks. Acaricidal control of ixodid ticks is impractical because of their wide distribution in forested areas, but houses infested with soft ticks can be sprayed with acaricidal formulations. Attached ticks should be removed without delay. The best way is to grasp the tick as close to the skin as possible with fine tweezers and pull firmly and steadily without twisting. Finally, despite the fact that most people who are bitten destroy the offending tick in disgust, it is recommended that they preserve specimens in ethanol for taxonomic identification and detection of pathogens by molecular methods.

Animals↗