一种收集蜱类唾液的方法

建立了一种收集蜱类唾液的方法。注射多巴胺(20mgmL)10μL于森林革蜱DermacentorsilvarumOlenev吸血雌蜱血腔内,可使唾液腺分泌唾液,用毛细管连续收集30min,可得唾液15～20μL蜱。     

Integrated Control of Acaricide-Resistant Boophilus Microplus Populations on Grazing Cattle in Mexico Using Vaccination with Gavac™ and Amidine Treatments

Throughout most of the twentieth century, tick infestations on cattle have been controlled with chemical acaricides, typically administered by dipping or spraying. This approach can cause environmental and residue problems and has created a high incidence of acaricide resistance within tick populations in the field. Recently we developed a vaccine against Boophilus microplus employing a recombinant Bm86 antigen preparation (Gavac), (Heber Biotec S.A., Havana, Cuba) which has been shown to induce a protective response in vaccinated animals. Here we show for the first time under field conditions a near 100% control of B. microplus populations resistant to pyrethroids and organophosphates, by an integrated system employing vaccination with Gavac and amidine treatments. This method effectively controls tick infestations while reducing the number of chemical acaricide treatments and consequently the rise of B. microplus populations resistant to chemical acaricides.     

Using genetics to control cattle parasites-the Rockhampton experience

Ever since their accidental introduction, cattle ticks, gastrointestinal nematodes and buffalo flies have been major parasites of cattle in northern Australia. Enormous effort and resources have been directed at chemical control of these parasites but the problem persists in undiminished form. The principal control measure remains the use of breeds that have some degree of parasite resistance. No breed is completely resistant and all are at times adversely affected by parasites. Complete resistance is the ultimate solution but has been generally ignored as a commercial reality. Studies at Rockhampton have demonstrated that completely resistant lines can be developed by genetic means within a commercially acceptable timeframe from even the most parasite-susceptible breeds. Genetic changes in tick and worm resistance over 15 years in response to selection for increased tick resistance in the Belmont Adaptaur (Bos taurus) line are reported. Costs and benefits of achieving increased tick resistance are examined and the applicability of the results to other breeds is discussed. Breed differences in resistance to buffalo flies and their effects on live weight are also reported and the possibility of selecting for increased buffalo fly resistance is explored.     

Chemical attraction of Dermacentor variabilis ticks parasitic to Peromyscus leucopus based on host body mass and sex

Macroparasites are commonly aggregated on a small subset of a host population. Previous explanations for this aggregation relate to differences in immunocompetence or the degree to which hosts encounter parasites. We propose active tick host choice through chemical attraction as a potential mechanism leading to aggregated tick burdens. We test this hypothesis using a Y-maze olfactometer, comparing chemical attraction responses of larval and nymphal Dermacentor variabilis ticks parasitic to the white-footed mouse, Peromyscus leucopus, as a function of host sex and host body mass. We hypothesized that larger hosts and male hosts would be most attractive to searching ticks, as these hosts commonly have higher tick burdens in the field. Chemical attraction trials were run in the presence and absence of a known tick attractant, host-produced carbon dioxide (CO₂). Male hosts and larger hosts were preferred by nymphal D. variabilis in the presence and absence of CO₂, whereas larvae had no detectable host preference. The current study suggests that host-produced chemical cues may promote aggregated tick burdens among hosts of a single species based on host body mass and sex.     

Methods currently used for the control of heartwater: their validity and proposals for future control strategies

Major advances made in the last few years in the knowledge of heartwater have not yet resulted in significant improvement of its prevention and control. Acaricide resistance and the high cost of chemical vector control stimulate research on alternative methods of vector control, such as immunization against the tick and the use of pheromones. Recently developed in vitro methods for the culture of Cowdria ruminantium may make it possible to arrive at safe recombinant or molecular vaccines. Specific chemotherapy may be improved by new laboratory screening methods, and studies on the pathogenesis of heartwater could lead to better non-specific treatment as well. Studies on the heritability of natural resistance to heartwater should determine whether or not practical selection and transfer of resistance is feasible. More research is also needed on the immunology and diagnosis of the disease and the factors involved in the resistance of young animals.     

Probing the functional role of tick metalloproteases

Tick saliva assists feeding through a complex mixture of compounds that disarm the host homeostasis processes, such as platelet aggregation, vasoconstriction and blood clotting, as well as innate and acquired immune responses. Although the various properties of tick salivary glands have sparked great interest as candidate sources for anti‐tick vaccines to prevent tick and tick‐borne diseases, antigens that can be useful to induce an immune response against tick bites or the pathogens transmitted by ticks have not yet been developed. Metalloproteases, which have been found in tick saliva, salivary gland, ovary and midgut, play an important role in inflammation, immunomodulation, fibrinolysis, blood protein digestion, nociception, vitellogenesis, remodelling of extracellular matrix and pathogen transmission. A large proportion of tick metalloproteases belong to the metzincin group, whose members characteristically have a highly conserved zinc‐binding motif integrated into the central α helix at the active site, and a methionine‐containing triad called Met‐turn followed by a cysteine‐rich domain at the C‐terminal site. This review discusses specifically the biological aspects of metalloproteases in tick physiology that have been published to date.     

Amblyomma americanum (L.): protein kinase C-independent fluid secretion by isolated salivary glands.

Protein kinase C activity was partially purified from tick salivary glands by fast protein liquid chromatography anion-exchange chromatography. Enzyme activity was stimulated by Ca2+, phosphatidylserine, and diacylglycerol with the highest activity observed in the presence of all three modulators. Enzyme activity was inhibited by a synthetic pseudosubstrate peptide with an amino acid sequence resembling the protein kinase C substrate phosphorylation site. The protein kinase C activator, 1-oleoyl-2-acetyl-sn-glycerol (OAG), when added to whole in vitro salivary glands previously prelabeled with 32P, stimulated the phosphorylation of salivary gland proteins. Activators of protein kinase C (phorbol ester or OAG) did not stimulate fluid secretion by isolated tick salivary glands. OAG and phorbol ester had only minimal affects on the ability of dopamine to stimulate secretion by isolated salivary glands and dopamine's ability to increase salivary gland cyclic AMP.     

Protein phosphorylation and control of tick salivary gland function

Tick salivary glands are controlled by nerves, dopamine being a neurotransmitter at the neuroeffector junction. Dopamine and cyclic AMP (cAMP) stimulate fluid secretion by isolated salivary glands. Dopamine activates an adenylate cyclase to increase intracellular cAMP within the female salivary glands. Phosphoproteins whose levels of phosphate are affected by cAMP-dependent protein kinase have been identified in subcellular fractions. Protein(s) phosphorylated by cAMP appears to activate protein phosphatase in the salivary glands.Another phosphorylation pathway appears to act through protein kinase C because of an ability of phorbol esters (known activators of protein kinase C) to stimulate the phosphorylation of proteins, and an ability of a peptide factor in tick brain to metabolize salivary-gland phosphoinositides, an event that often precedes activation of protein kinase C. Because cAMP modulates brain-factor-stimulated formation of inositol phosphates (products of phosphoinositide breakdown) an interrelationship between the two pathways seems likely.Evidence of regulatory processes, including protein phosphorylation'dephosphorylation reactions, will provide a basis for helping asses the physiological significance of secretory products and the role of the salivary glands in disease transmission.     

Perturbation of tyraminergic/octopaminergic function inhibits oviposition in the cattle tick Rhipicephalus (Boophilus) microplus

The cattle tick Rhipicephalus microplus, is one of the most damaging livestock ectoparasites. Tropical tick infestation limits the introduction of high-yield bovine varieties because they do not have immunity to the diseases transmitted by these ectoparasites. This tick is usually controlled with chemical acaricides but their indiscriminate use has created resistant populations. The discovery of new molecules that can be used for tick control is urgent. Based on the knowledge that octopamine, a biogenic amine analog to epinephrine, is central to the regulation of oviposition in several studied arthropods and that an imbalance in octopamine release causes sterility in a Drosophila model. Tyramine, octopamine and epinastine and 83 adrenergic compounds classified by their effect in the vertebrate systems were screened for their ability to block oviposition in Rhipicephalus microplus. Of these molecules, we found that 10 alpha-agonists, 3 alpha-antagonists, 5 beta-adrenergic agonists, 7 beta-antagonists and Norepinephrine were able to inhibit oviposition in this tick at pharmacological concentrations. Surprisingly, tyramine appears to be more potent than octopamine. The probable physiological causes of this inhibition are discussed. Our results suggest that although there are alpha adrenergic-like receptors in the tick, they do not behave in a manner completely analogous to their vertebrate counterparts.     

Control of fluid secretion by isolated salivary glands of the lone star tick.

Adrenaline, noradrenaline, cyclic-AMP, and theophylline stimulated salivary fluid secretion in glands isolated from rapidly engorging females of the lone star tick, Amblyomma americanum. Cyclic-AMP and theophylline effectively initiated secretion only after the glands were pre-incubated and pre-stimulated to secrete with adrenaline. Pilocarpine nitrate, 5-hydroxytryptamine (5-HT), and glutamate did not stimulate glandular secretion, 2,4-Dinitrophenol (DNP) slowed adrenaline's ability to stimulate secretion. Results obtained after additions of harmaline and ouabain point to the possibility of a (Na⁺+K⁺)-ATPase system for helping to move fluid across the glandular epithelium. The significance of these findings to the mechanisms and control of tick salivary glandular function is discussed.     

The history of the cattle tick Boophilus microptus in Australia and achievements in its control

For more than 100 years, tick fever and the cattle tick have caused tremendous financial loss to cattle producers around the world. Since Australia became infected with the disease and infested with its tick vector in the mid-19th century, a great deal of research effort has been directed towards their effective control by Australian farmers, administrators and scientists. Such research has yielded information which has facilitated the development of various control strategies that have equal application in other countries afficted with the same problem. It has been demonstrated that integration of a variety of these strategies is necessary for long-lasting control.     

Perspectives on the potential of entomopathogenic fungi in biological control of ticks

Ticks are serious health threats for humans, and both domestic and wild animals. Ticks are controlled mostly by application of chemical products; but these acaricides have several negative side effects, including toxicity to animals, environmental contamination, and induction of chemical resistance in some tick populations. Entomopathogenic fungi infect arthropods in nature and can occur at enzootic or epizootic levels in their host populations. Laboratory studies clearly demonstrate that these fungi can cause high mortality in all developmental stages of several tick species, and also reduce oviposition of infected engorged females. Tick mortality following application of fungi in the field, however, often is less than that suggested by laboratory tests. This is due to many negative biotic and climatic factors. To increase efficacy of fungal agents for biological control of ticks under natural conditions, several points need consideration: (1) select effective isolates (viz., high virulence; and tolerance to high temperature, ultraviolet radiation and desiccation); (2) understand the main factors that affect virulence of fungal isolates to their target arthropods including the role of toxic metabolites of the fungal isolates; and (3) define with more precision the immune response of ticks to infection by entomopathogenic fungi. The current study reviews recent literature on biological control of ticks, and comments on the relevance of these results to advancing the development of fungal biocontrol agents, including improving formulation of fungal spores for use in tick control, and using entomopathogenic fungi in integrated pest (tick) management programs.     

Hard tick calreticulin (CRT) gene coding regions have only one intron with conserved positions and variable sizes

Calreticulin (CRT) is a unique eukaryotic gene. The CRT gene product, calreticulin, was first identified as a calcium binding protein in 1974, but further investigations have indicated that CRT protein performs many functions in cells, including involvement in evading the host's immune system by parasites. Many studies of CRT have been published since the molecule was first discovered; however, the CRT gene exon-intron structure is only known for a limited number of ectoparasite species. In this study, we compared tick CRT genomic sequences to the corresponding cDNA from 28 species and found that 2 exons and 1 intron are present in the tick CRT gene. The intron position is conserved in 28 hard ticks, but intron size and nucleotide sequences vary. Three tick introns possess duplicated fragments and are twice as long as other introns. All tick CRT introns obey the GT-AG rule in the splice-site junctions and are phase 1 introns. By comparing tick CRT introns to those of fruit fly, mouse, and human, we conclude that tick CRT introns belong to the intron-late type. The number and size of CRT introns have increased through the evolution of eukaryotes.     

Association of BoLA-DRB3.2 alleles with tick (Boophilus microplus) resistance in cattle

Losses caused by bovine tick burdens in tropical countries have a tremendous economic impact on production systems. Besides reducing production, this parasite can cause death in the most susceptible animals. The use of commercial acaricides has been the major method of control, but their misuse has led to tick resistance to many chemicals. More recently, vaccines have been used in some countries without solving the problem completely. An alternative could be the development of resistant animals and the use of genetic markers and candidate genes that could help with the enormous task of selecting resistant animals. The bovine lymphocyte antigen genes (BoLA) have been shown to be associated with some parasitic infestations and disease incidence. Thus, the objective of the present study was to determine the association of BoLA-DRB3.2 alleles with tick resistance in cattle. The study was conducted on 231 F2 (Gyr x Holstein) animals that were artificially infested with 10,000 tick larvae. Log of tick count +1 was used as the dependent variable in a mixed animal model with allele substitution effects in addition to fixed effects of year and season at tick count, sex of calves, age of animal at tick count, hair type (short-straight, short-curl, long-straight, and long-curl), coat color (white, >75% white, 50- 75% white, and 25-50% white), and additive genetic, permanent environmental and residual effects as random. Females showed fewer ticks than males. Animals with short-straight hair were more resistant to tick infestation than animals with long-curl hair, and animals with whiter coat color also had fewer ticks. An association between BoLA alleles and lower tick number was found for alleles DRB3.2 *18, *20 and *27 at the 5% significance level. Also, one allele (DRB3.2*16) showed an association at the 10% level. Allele *27 was the most frequent in the population (30.7%), followed by alleles *16 (10.8%), *20 (8.7%) and *18 (2.4%). These results suggest that BoLA-DRB3.2 alleles could be used to help in the selection of animals resistant to tick infestation. However, further studies involving a larger population of cattle in combination with other BoLA genes may help to understand the mechanisms of resistance to parasites.     

Responses of cattle to allergens from Boophilus microplus

Extracts of larvae of the cattle tick, Boophilus microplus, contain two allergens giving immediate hypersensitivity reactions in the skin of cattle which have been exposed to the tick. The partial purification of one of these is described. This, together with the other allergen whose purification has been described previously, was used to investigate the relationships between tick resistance, immediate hypersensitivity reactions, reaginic antibody levels as measured by Prausnitz-Küstner type reactions and serum levels of agglutinating antibody to one of the allergens. There were significant correlations between resistance and both the immediate hypersensitivity reactions and agglutinating antibody levels.     

Suppression of Th2 cytokines reduces tick-transmitted Borrelia burgdorferi load in mice

Previous work has indicated that both Borrelia burgdorferi and the process of tick feeding (saliva) modulate the host immune response. Molecules have been identified in tick saliva that effect T cell proliferation by binding to specific cytokines, thereby promoting a Th2 cytokine response that does not afford protection against tick-transmitted B. burgdorferi in mice. Moreover, reconstitution of a Th1-biased T cell response prior to spirochete challenge effectively neutralizes tick modulation of host immunity and affords protection against tick transmission of spirochetes. The current studies were undertaken to determine the effect of neutralizing specific Th2 cytokines prior to tick feeding and subsequent transmission of B. burgdorferi. The results indicate that suppression of both IL-4 and IL-5 prior to the feeding of B. burgdorferi-infected ticks significantly decreased spirochete load in target organs such as joint, bladder, heart, and skin of the Lyme disease-susceptible host.     

Exon-intron structure of outlier tick lipocalins indicate a monophyletic origin within the larger lipocalin family

All tick proteins assigned to the lipocalin family lack the structural conserved regions (SCRs) that are characteristic of the kernel lipocalins and can thus be classified as outliers. These tick proteins have been assigned to the tick lipocalin family based on database searches that indicated homology between tick sequences and the fact that the histamine binding protein (HBP2) from the hard tick Rhipicephalus appendiculatus (Ixodidae) shows structural similarity to the lipocalin fold. Sequence identity between kernel and outlier lipocalins falls below 20% and the question raised is whether the outlier and kernel lipocalins are truly homologous. More specifically in the case of the tick lipocalins, whether their structural fold is derived from the lipocalin fold or whether convergent evolution resulted in the generation of the basic lipocalin-like fold which consists of an eight stranded continuous anti-parallel beta-barrel terminated by a C-terminal alpha-helix that lies parallel to the barrel. The current study determined the gene structure for HBP2 and TSGP1, TSGP2 and TSGP4, lipocalins identified from the soft tick Ornithodoros savignyi (Argasidae). All tick lipocalins have four introns (A-D) with conserved positions and phases within the tick lipocalin sequence alignment. The positions and phase information are also conserved with regard to the rest of the lipocalin family. Phylogenetic analysis using this information shows conclusively that tick lipocalins are evolutionary related to the rest of the lipocalin family. Tick lipocalins are grouped within a monophyletic clade that indicates a monophyletic origin within the tick lineage and also group with the other arthropod lipocalins in a larger clade. Phylogenetic analysis of sequence alignments based on conserved secondary structure of the lipocalin fold support the conclusions from the gene structure trees. These results indicate that exon-intron arrangement can be useful for the inclusion of outlier lipocalins within the larger lipocalin family.     

Using agent-based models to inform the dynamics of winter tick parasitism of moose

North American moose (Alces alces) populations along the southern extent of their range have been experiencing high levels of calf mortality in recent years. In New England, this phenomenon has been linked to extensive blood loss resulting from extreme winter tick (Dermacentor albipictus) parasitism. Moose are symbolic of the region and generate income through tourism and the auction of hunting permits; thus, successive years of greater than 50% calf mortality (epizootics) are of concern to wildlife managers and others. It is hypothesized that high localized moose density coupled with climate change are the driving forces behind moose-winter tick epizootics; however, the impact that variable combinations of these and other factors have on the occurrence and magnitude of epizootics has not been previously simulated. We, therefore, developed and implemented a spatially explicit agent-based model with two model environments, each representative of a distinct location within an ongoing field study site in northern New Hampshire that differed in the proportional availability of optimal moose habitat. Three experiments were devised to test the sensitivity of the outcome variables, calf infestation level and calf mortality, to 1) winter tick abundance, 2) winter tick aggregation, and 3) moose density, for the length of the winter tick questing period. Each experiment was conducted in both model environments. This model generated similar mortality levels to those measured in the field study under representative moose density and weather conditions. Additionally, the modeled moose agents and the radio-collared moose in the field reflect comparable habitat usage. While the infestation levels reported by calf agents are considered conservative, we believe that future versions of this model, parameterized with more accurate estimates of winter tick abundance and moose density, may be an effective tool for managing moose against winter tick parasitism.     

Amblyomma americanum: identification of tick salivary gland antigens from unfed and early feeding females with comparisons to Ixodes dammini and Dermacentor variabilis

Salivary gland antigens involved in host resistance to tick feeding by Amblyomma americanum (lone star tick) have been identified. Gland extracts from unfed and partially fed 12-, 48-, 72-, 96-, and 120-hr females and their corresponding midgut tissues were analyzed by immunoblotting with sera from naturally immune and hyperimmune sheep and rabbits. Polypeptides at 90, 75, 58, 45, 33, and 23 kDa from the salivary glands of A. americanum females were consistently observed with antibodies from both sheep and rabbits. No antigens unique to tick midgut tissue were detected with immune sera. Female Dermacentor variabilis and Ixodes dammini shared 90- and 45-kDa salivary gland antigens with A. americanum, and these may represent conserved polypeptides. We speculate that some of the salivary gland antigens represent components of tick cement, while others are playing some other yet undetermined role in tick feeding.