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Expression of defensin-like peptides in tick hemolymph and midgut in response to challenge with Borrelia burgdorferi, Escherichia coli and Bacillus subtilis.

Challenge of Dermacentor variabilis by hemocoel injection with Borrelia burgdorferi but not Bacillus subtilis or Escherichia coli provoked secretion of two low molecular weight peptides into the hemolymph plasma; the lower band co-migrated with a band previously identified as varisin (a tick defensin). These findings are consistent with reports that D. variabilis controls B. burgdorferi but not B. subtilis or E. coli by defensin-dependent bacteriolysis. Challenge of the tick midgut by capillary artificial feeding with bacteria also provoked expression of multiple low molecular weight peptides. In this case, however, all three bacteria elicited the response. Two bands, including the defensin-like peptide were expressed following challenge with B. subtilis and E. coli, but only the upper band following challenge with B. burgdorferi. Although they appeared intact, these spirochetes were no longer viable suggesting that borreliae in the midgut are controlled by a different method than the lytic response of the D. variabilis hemolymph. DD-RT-PCR revealed multiple mRNAs in the midgut of D. variabilis following challenge with B. burgdorferi, E. coli and Rickettsia montana. Although their identification remains to be determined, the large number of genes expressed in response to bacterial challenge presents intriguing possibilities for explaining the ability of the tick midgut to destroy invading microbes at the cellular level.

Animals↗

Steinernema glaseri Santa Rosa strain (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora CCA Strain (Rhabditida: Heterorhabditidae) as biological control agents of Boophilus microplus (Acari: Ixodidae).

The present study was carried out to evaluate the action of Steinernema glaseri Santa Rosa strain and Heterorhabditis bacteriophora CCA strain as biological control agents of Boophilus microplus. Engorged females ticks were distributed on Petri dishes containing different concentrations of infective juvenile (IJ) nematodes (0, 375, 500, 750, 1,500, 2,500, 5,000 and 25,000). The data showed a reduction of approximately 90% in the eggs laid at a concentration of 5,000 S. glaseri IJs and approximately 80% at a concentration of 1,500 H. bacteriophora IJs. The female mortality increased linearly with the increase in S. glaseri concentrations. However, in the tests with H. bacteriophora this linearity was not observed. The effectiveness of the treatment with both species of entomopathogenic nematodes was compatible with other control methods. The results show the potential of S. glaseri and H. bacteriophora as biological control agents for the control of B. microplus under laboratory conditions.

Animals↗

Persistence of Hepatozoon americanum (Apicomplexa: Adeleorina) in a naturally infected dog.

To determine the persistence of Hepatozoon americanum in a naturally infected dog, skeletal muscle biopsies were performed at approximately 6-mo intervals over a period of 5.5 yr, and the samples were examined for presence of lesions of American canine hepatozoonosis (ACH). Nymphal Amblyomma maculatum (Gulf Coast tick) were allowed to feed to repletion on the dog periodically over the 5.5-yr period, and adult ticks were dissected and examined for presence of H. americanum oocysts. With 3 exceptions, the biopsied muscle contained lesions characteristic of ACH; no evidence of infection was found at 36, 54, and 67 mo after the original diagnosis. In every instance, nymphal Gulf Coast ticks became infected, indicating that dogs naturally infected with H. americanum can remain infectious for Gulf Coast ticks for at least 5.5 yr. Skeletal muscle biopsy is a reasonably reliable method of determining whether dogs are infected with the parasite. Xenodiagnosis using nymphal Gulf Coast ticks is an even more sensitive method, but the procedure is practicable only experimentally. Design of prevention and control measures for ACH must take into account knowledge that the parasite can survive in dogs, and presumably other vertebrate host(s), for long periods. Preventing ingestion of Gulf Coast ticks is an effective control measure.

Animals↗

Combined prime-boost vaccination against tick-borne encephalitis (TBE) using a recombinant vaccinia virus and a bacterial plasmid both expressing TBE virus non-structural NS1 protein.

BACKGROUND: Heterologous prime-boost immunization protocols using different gene expression systems have proven to be successful tools in protecting against various diseases in experimental animal models. The main reason for using this approach is to exploit the ability of expression cassettes to prime or boost the immune system in different ways during vaccination procedures. The purpose of the project was to study the ability of recombinant vaccinia virus (VV) and bacterial plasmid, both carrying the NS1 gene from tick-borne encephalitis (TBE) virus under the control of different promoters, to protect mice against lethal challenge using a heterologous prime-boost vaccination protocol. RESULTS: The heterologous prime-boost vaccination protocol, using a VV recombinant and bacterial plasmid, both containing the NS1 TBE virus protein gene under the control of different promoters, achieved a high level of protection in mice against lethal challenge with a highly pathogenic TBE virus strain. No signs of pronounced TBE infection were detected in the surviving animals. CONCLUSION: Heterologous prime-boost vaccination protocols using recombinant VV and bacterial plasmids could be used for the development of flavivirus vaccines.

Animals↗

Eradication and surveillance of the tropical bont tick in the Caribbean. An international approach.

The Caribbean Amblyomma program (CAP) is a multidonor funded effort involving several technical organizations and national governments in the eradication of the tropical bont tick. The Food and Agriculture Organization (FAO) is the lead technical agency, providing both technical and administrative support to the CAP Regional Coordination Unit (RCU) based in Barbados. All collaborating agencies, donors, and national representatives are members of the Amblyomma Program Council (APC), the overall governing body of the CAP. In addition to FAO, the other organizations involved are the Caribbean Community (CARICOM), the European Union (EU), the Inter-American Institute for Cooperation on Agriculture (IICA), the International Fund for Agricultural Development (IFAD), and the United States Department of Agriculture (USDA). Several agencies (EU, FAO, IFAD, and USDA) are also major donors. The CAP cooperates with the complementary Plan POSEIDOM Veterinaire Program in the French West Indies. The University of the West Indies (UWI), the International Institute for Biological Control (IIBC), and the Departement d'Elevage et de Medecine Veterinaire des Pays Tropicaux (EM-VT) in Guadeloupe are other collaborators.

Agriculture↗

Passive transfer of resistance in rabbits infested with adult Ixodes ricinus L: humoral factors influence feeding and egg laying.

Parital immunity against the bites of female I. ricinus was transferred to normal rabbits by inoculating immune serum from resistant animals. Transferred humoral factors diminished the weight of the ticks' blood meal by 29% and increased the feeding period by about 1 day in comparison with ectoparasites engorged on controls. They provoked also the failure of egg laying by female I. ricinus. Only 55% of ticks fed on treated rabbits laid eggs (94% on controls). The immunological state of immune serum donors or recipients was studied and the IgG and homocytotropic specific anti-I. ricinus antibodies were identified. The immediate hypersensitivity of rabbits' skin was also controlled.

Animals↗

Epidemiological aspects of human granulocytic Ehrlichiosis in southern Germany.

Human granulocytic Ehrlichiosis (HGE) is a newly emerging acute febrile illness which is likely transmitted by ticks of the Ixodes ricinus/I. persulcatus complex. First seroepidemiological surveys on the prevalence of HGE antibodies, detection of DNA of granulocytotropic Ehrlichiae in I. ricinus and one case of HGE from Slovenia confirmed by serology and PCR (polymerase chain reaction) suggest that HGE might exist all over Europe. The purpose of the present study was a) to determine the prevalence of antibodies against the HGE agent in sera collected from persons at high risk for exposure to I. ricinus with that of a control population and b) to determine the prevalence of granulocytic Ehrlichiae in I. ricinus ticks from Southern Germany. We studied sera from 150 forestry workers and 105 patients with an established diagnosis of Lyme disease as tick-exposed populations. Sera from 103 healthy blood donors without a history of known tick bites served as controls. A significantly higher prevalence of HGE antibodies (P < or = 0.01) was present among patients with Lyme borreliosis (12 of 105 were positive; 11.4%) and forestry workers (21 of 150 were positive; 14%) compared to blood donors (2 of 103 were positive; 1.9%). Furthermore, 510 adult and nymphal I. ricinus were investigated by PCR for the presence of granulocytic Ehrlichiae with primers specific for the E. phagocytophila group. In eight (1.6%) of the investigated ticks the expected amplification product was detectable, indicating a low prevalence of infected ticks especially when compared with B. burgdorferi. The presented data strongly suggests that the HGE agent or a closely related organism exists in Southern Germany and therefore HGE should be considered in the differential diagnosis of febrile illnesses. However, final evidence can be provided only after isolation of the organism from patients.

Adult↗

Entomopathogenic potential of Metarhizium anisopliae isolated from engorged females and tested in eggs and larvae of Boophilus microplus (Acari: Ixodidae).

The purpose of this work was to evaluate the in vitro virulence of three isolates of Metarhizium anisopliae var. anisopliae to eggs and larvae of the tick Boophilus microplus. The fungus tested was isolated from engorged females of B. microplus collected in the field, and identified as Ma01, Ma02 e Ma04. These isolates were evaluated by immersion of eggs and larvae in suspension with different conidial concentrations: 10(5), 10(6), 10(7) e 10(8) conidia/ml. In each isolate there was a treatment group for each spore concentration and a control group with 10 repetitions. It was observed in the treated egg groups that there was a hatching percentage that was much less than that observed in the control groups. This was in inverse proportion to the conidia concentration/ml. Larval bioassays of all the tested isolates resulted in a high mortality of larvae in direct proportion to the spore concentration/ml, 10 days after the conidia suspensions were inoculated. To consolidate the infection, the fungus used in bioassays was re-isolated.

Animals↗

Interbreed differences in the efficacy of 1% deltamethrin pour-on to protect small livestock against infestation with Ixodes rubicundus (Acari:Ixodidae).

Synthetic pyrethroid pour-on compounds have been developed for use on cattle but many are now also registered for the control of ectoparasites on small ruminants. The breed of small livestock has been known to influence the efficacy of various pour-ons and we hypothesized that differences in the duration of protection afforded by a synthetic pyrethroid pour-on against ticks on small livestock, would be greatest between hairy and woolly breeds. Angora goats with mohair, Dorper sheep with hairy wool, and Merino sheep with wool were treated with 1% m/v deltamethrin pour-on with an oil base at a dosage rate of 1 ml 5 kg-1 body mass. The duration of effective protection against Ixodes rubicundus, the Karoo paralysis tick, was determined under field conditions. Angora goats proved to be the highest risk group, but were afforded the least protection. Efficacy never reached 100% in this group and within 3 weeks the goats were already at risk of paralysis. Dorper sheep were afforded above 90% protection for 3 weeks and 85% after 4 weeks. In the case of the Merino sheep efficacy of protection remained above 90% up to 7 weeks post-treatment and was still 87% during Week 8. Differences in the amount of sebum and suint in the epidermis of the different breeds may be responsible for differences in the rate of spread and the rate of dilution of the active compound in the epidermis.

Administration, Topical↗

[Decreased antioxidant-defence mechanisms in cerebrospinal fluid (CSF) in patients with tick-borne encephalitis (TBE)].

14 patients with Tick-borne Encephalitis (TBE) aged 21-64 (mean = 42.3) were analysed. The activity of superoxide dismutase (SOD), glutathione reductase (GSSG-R), glutathione peroxidase (GSH-Px), concentrations of malondialdehyde (MDA) and total sulphydryl groups (-SH) were measured in cerebrospinal fluid (CSF). Control group consisted of 10 patients whose CSF parameters remained in normal range. The CSF examination was performed twice: before and 3 weeks after treatment. The analysed activity of SOD, GSH-Px, GSSG-R, MDA and total sulphydryl groups (-SH) during the acute stage of the disease was significantly lower comparing to the control group. Despite the treatment, GSSG-R activity, MDA concentration and total sulphydryl groups--SH further lowered significantly. Although the SOD activity in CSF was higher in the second examination, it remained significantly lower comparing to the control group. We showed that the GSH-Px and GSSG-R activity in CSF after the acute stage of the TBE remained significantly lower than in the control group. Our examinations prove that during the TBE an increased generation of oxygen-derived free radicals occurs what shows decreased activity of the antioxidant parameters (SOD, GSH-Px, GSSG-R) and decreased concentration of total sulphydryl groups--SH in CSF. Our results suggest that during TBE, molecular structures injury of enzymes and antioxidative reactive cofactors may occur.

Adult↗

Vaccines for preventing tick-borne encephalitis.

BACKGROUND: Tick-borne encephalitis is a disease of the central nervous system caused by a virus. Other than the vaccine, there is no treatment for the disease. OBJECTIVES: The objective of this review was to assess the effects of vaccines to prevent tick-borne encephalitis. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Vaccine Fields Trials Register, the Cochrane Controlled Trials Register, Medline, Embase and reference lists of articles. We also handsearched the journal Vaccine. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing tick-borne encephalitis vaccines against placebo, control vaccines or comparisons of different doses or schedules of tick-borne encephalitis vaccines. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied inclusion criteria. A panel of six assessors examined trial quality. MAIN RESULTS: Five trials were included. They could not be combined for meta-analysis because of differences in comparisons and outcomes. Four types of tick-borne encephalitis vaccines were used. All the vaccines gave seroconversion rates of over 87%. There were frequent reports of systemic and local adverse effects. REVIEWER'S CONCLUSIONS: Tick-borne encephalitis vaccines appear to be highly immunogenic, but the relationship between seroconversion and clinical protection has not been established. Although adverse effects were commonly reported, none were severe or life threatening.

Child↗

The efficiency of patch sampling for determination of relative tick burdens in comparison with total tick counts.

Quantitative data on host tick burdens are fundamental for the initiation of control strategies and effective management of wildlife populations, but the methods of live sampling employed for domestic animals are unsuitable for sampling wild animals. Despite advances in the use of destructive methods (the scrub and digestion techniques) to obtain measures of the total tick burden on wildlife, these methods are too involved for many field workers, who often need only measures of relative tick burden. Recently, patch sampling methods have been introduced whereby only certain predilection sites are sampled, the presumption being that the number of ticks collected gives an indication of the relative degree of infestation. We examined the validity of patch sampling as a measure of relative tick burden by comparing adult ticks collected from the ears, head, neck, foreleg and perianal region of impala (Aepyceros melampus) with total tick burdens of the same animals derived from the digestion technique. Adult ticks from patch sampling were positively and significantly correlated with total adults and total ticks (larvae, nymphs, and adults) on impala, with ticks patch sampled from the neck showing the highest correlation with the total tick burden. Comparison of relative tick loads from patch sampling with absolute tick loads from digestion for three classes of impala (females, bachelor males and territorial males) gave qualitatively similar results. We conclude that, when measures of relative tick load are sufficient and destructive sampling is not feasible, patch sampling can provide reliable information on relative tick burdens that are positively correlated with the total tick burden.

Animals↗

Effects of temperature on development and growth in the tick, Haemaphysalis longicornis.

As a part of ecological studies on Haemaphysalis longicornis, the effects of controlled temperatures (12, 15, 20, 25 and 30 degrees C; approximately 100% RH) on development and growth of the tick were investigated and the critical low temperature for each stage in the life cycle was estimated. As the temperature became low, the periods of preoviposition, oviposition, egg hatching (incubation) and moulting were prolonged. At 12 degrees C, however, oviposition, egg hatching and moulting of the larva and nymph did not occur. The critical low temperatures for oviposition, egg hatching (developmental zero) and larval and nymphal moulting, which were calculated theoretically from the regression equations, were 11.1, 12.2, 10.2 and 11.8 degrees C, respectively. The temperature also affected the egg productivity and hatch-ratio. The number of deposited eggs per mg of body weight decreased markedly at 15 degrees C, and the hatch-ratio was lowered with dropped temperatures.

Animals↗

Identification of functional promoters in the msp2 expression loci of Anaplasma marginale and Anaplasma phagocytophilum.

Organisms in the family Anaplasmataceae are important tick-borne pathogens of livestock worldwide and cause recently emergent infections in humans. Despite their medical importance, very little is known about how these organisms regulate gene expression in the mammalian host, the tick vector, or during transition between the host and vector. However, it is clear that gene regulation, in addition to recombinatorial mechanisms, is essential for these small genome pathogens to adapt to distinctly different environments. In this study, we identify and establish the function of three promoter elements in the locus encoding major outer membrane protein expression sites in both Anaplasma marginale and Anaplasma phagocytophilum. Gene expression from this locus involves both classical and atypical polycistronic transcripts. The identified promoter elements have a structure similar to that defined in Escherichia coli and are functional in driving protein expression in a prokaryotic cell-free transcription and translation system and in recombinant E. coli. The two strongest promoters identified in vitro and with recombinant E. coli were also shown to be functional in A. marginale infected cells, as determined by quantification of downstream transcripts. The promoters in both A. marginale and A. phagocytophilum have similar structure and activity, supporting the conclusion that the two loci are syntenic with conservation of function. In addition, they share structural elements within the promoters that appear to be likely sites for regulation. These data enhance our understanding of how expression of these variable outer membrane proteins may be controlled in the key stages of tick-borne transmission and infection.

Animals↗

Strategies to interrupt the development of Anaplasma marginale in its tick vector. The effect of bovine-derived antibodies.

Anaplasma marginale is a rickettsia transmitted by ticks that invades and multiplies in bovine erythrocytes causing the disease anaplasmosis. A complex developmental cycle occurs within ticks that begins in midgut cells, with subsequent infection in gut muscle cells. Final development occurs in salivary glands from where the rickettsia is transmitted to the vertebrate host. At each site of development, A. marginale multiplies within membrane-bound inclusions. Attempts to control anaplasmosis have focused on cattle and have included immunization and prophylactic treatment with tetracyclines. New strategies for control of anaplasmosis are being focused on the tick vector. Development of vaccines against hemoparasites in ticks may be feasible because vertebrate host immunoglobulins appear to cross the midgut epithelium of invertebrates and enter the hemolymph without breakdown. We tested the effect of A. marginale antibodies ingested by ticks with the bloodmeal on infections in ticks. Cattle were immunized with purified outer membrane proteins of erythrocytic-derived parasites. Infections in ticks exposed to the immunized cattle were determined using an Anaplasma-specific DNA probe, light and electron microscopy, and tick transmission studies. Vaccine-derived antibodies did not appear to affect the development and transmission of A. marginale in ticks. Further studies are needed to determine if bovine antibodies remain intact within ticks and whether the tick stage of A. marginale has unique surface antigens from the erythrocytic stage.

Anaplasma↗

Immunization of cattle against Boophilus annulatus ticks using adult female tick antigen.

Ten crossbred calves (Friesian x Egyptian native) aged between 7-9 months old were used to evaluate the immunizing effect of whole adult B. annulatus tick antigen. A soluble fraction was obtained from extensively disrupted ticks and this fraction was used to vaccinate calves with 1 mg protein antigen. The vaccination protocol for five randomly selected calves involved two immunizations, the first was administered subcutaneously (plus aluminium hydroxide adjuvant) at the beginning of the experiment and the second was given 4 weeks later. At the same time, calves of the control group were injected with phosphate buffered saline (PBS) plus adjuvant. Ticks were counted on animals and the number of eggs laid per tick were counted and hatching percentages were determined. The vaccinated and control animals were skin tested with the antigen and calves were injected in three different sites with 50 microliters of 50, 100 and 200 ng of the antigen. At the same time, control sites were injected with PBS and the diameter of the immune response sites was measured using skin callipers. Immunization of calves showed that vaccination resulted in 73% reduction in the mean tick count. At the same time, immunization reduced the oviposition of ticks from vaccinated calves and the reduction percentage in egg laying was 65%. Vaccination induced a good immunity that could protect calves during the tick season (more than 5 months) as indicated from skin hypersensitivity reaction with the tick antigen.

Animals↗