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Production and characterization of monoclonal antibodies against midgut of ixodid tick, Haemaphysalis longicornis.

There are concerted efforts toward development of tick vaccines to replace current chemical control strategies that have serious limitations [Parasitologia 32 (1990) 145; Infectious Disease Clinics of North America (1999) 209-226]. In this study, monoclonal antibodies (mAbs) specific to Haemaphysalis longicornis midgut proteins were produced and characterized. Eight antibody-secreting hybridomas were cloned and the mAbs typed as IgG1, IgG2a and IgG2b. On immunoblots, all mAbs reacted with a midgut protein band of about 76 kDa. All mAbs uniformly immunogold-stained the surface or epithelial layers of H. longicornis midgut and endosomes. Adult ticks (50%) that fed on an ascitic mouse producing the IgGs developed a red coloration and did not oviposit. As such, the 76 kDa protein that reacted with the mAbs could, therefore, be a potential candidate for tick vaccine development.

Animals↗

Antibody to a cDNA-derived calreticulin protein from Amblyomma americanum as a biomarker of tick exposure in humans.

The antibody responses of human and animal hosts were studied to determine the utility of antibody against recombinant tick calreticulin (rTC), a cDNA-derived protein isolated from salivary glands of Amblyomma americanum L., as a biologic marker of tick exposure. Rabbits fed upon by either A. americanum or Dermacentor variabilis Say developed significant anti-rTC antibody responses, as measured by both ELISA and immunoblot assay. In contrast, gerbils exposed to Aedes aegypti did not develop anti-rTC antibodies, as measured by ELISA or immunoblot assay. The utility of the assay was next evaluated in humans at high risk for tick exposure. During April through September 1990, 192 military personnel who originated from either Fort Chaffee, Arkansas or Fort Wainwright, Alaska were studied during maneuvers in tick infested areas at Fort Chaffee. Study subjects completed a questionnaire and had pre- and post-maneuvers serum specimens analyzed for antibodies to rTC. In adjusted analysis (controlling for age, fort of origin, attached tick during maneuvers, and bed netting use), the use of bed netting and home station were associated with post-maneuvers anti-rTC antibody seropositivity by ELISA. Subjects from Fort Wainwright were more likely to be seropositive for anti-rTC antibody (adjusted odds ratio = 5.3, 95% confidence interval [CI] = 1.1-25.6). Personnel who did not report the use of bed netting were more likely to be anti-rTC seropositive (adjusted odds ratio = 6.8, 95% CI = 1.4-32.4). Immunoblot assays showed that humans had specific anti-rTC antibody responses. The animal experiments demonstrate that hosts exposed to naturally feeding ticks develop anti-rTC antibodies. The data also indicate that hosts exposed to Ae. aegypti saliva may not develop antibodies against rTC. Observations in tick-exposed humans support the hypothesis that anti-rTC antibody seropositivity is a biologic marker of tick exposure.

Adult↗

[Ability of female ixodid ticks to recover from poisoning with DDT in the process of feeding].

Ability of hungary females of Ixodes persulcatus, Haemaphysalis concinna and Dermacentor silvarum to feed and lay eggs after having been exposed to different DDT doses (LD100) has been studied. The ticks heavily injuried but still mobile can feed on the host and lay eggs, i.e. to overcome the poisoning. Clutches laid by these females do not differ from control ones. Loss of mobility deprives the ticks of the ability to fed. An increase in DDT dose (LD100 X 100, experiment with I. persulcatus) does not allow the ticks to feed even if the ability to movement is preserved.

Animals↗

Pathogenicity of Steinernema carpocapsae and S. glaseri (Nematoda: Steinernematidae) to Ixodes scapularis (Acari: Ixodidae).

The entomopathogenic nematodes Steinernema carpocapsae (Weiser) and S. glaseri (Steiner) are pathogenic to engorged adult, blacklegged ticks, Ixodes scapularis (Say), but not to unfed females, engorged nymphs, or engorged larvae. Nematodes apparently enter the tick through the genital pore, thus precluding infection of immature ticks. The timing of tick mortality, and overall mortality after 17 d, did not differ between infections by S. carpocapsae and S. glaseri. These nematodes typically do not complete their life cycles or produce infective juveniles in I. scapularis. However, both species successfully produced infective juveniles when the tick body was slit before nematode infection. Mortality of engorged I. scapularis females infected by S. carpocapsae was greater than uninfected controls, but did not vary significantly with nematode concentration (50-3,000 infective juveniles per 5-cm-diameter petri dish). The LC50 was 347.8 infective juveniles per petri dish (5 ticks per dish). Hatched egg masses of infected ticks weighed less than those of uninfected controls. Mortality of infected ticks was greatest between 20 and 30 degrees C, and was lower at 15 degrees C.

Animals↗

Vaccination of cattle with Anaplasma marginale derived from tick cell culture and bovine erythrocytes followed by challenge-exposure with infected ticks.

Anaplasmosis, a hemolytic disease of cattle caused by the tick-borne pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae) has been controlled using killed vaccines made with antigen harvested from infected bovine erythrocytes. We recently developed a cell culture system for propagation of A. marginale in a continuous tick cell line. In this study, we performed a cattle trial to compare the bovine response to vaccination with A. marginale harvested from tick cell culture or bovine erythrocytes. All immunized and control cattle were then challenge-exposed by allowing male Dermacentor variabilis infected with A. marginale to feed and transmit the pathogen. Nine yearling cattle (three per group) were used for this study and were immunized with cell culture-derived A. marginale, erythrocyte-derived A. marginale or received adjuvant only to serve as controls. Each vaccine dose contained approximately 2 x 10(10) A. marginale and three immunizations were administered at weeks 1, 4 and 6. At week 8, cattle were challenge-exposed by allowing 60 D. variabilis male that were infected with A. marginale as adults to feed on the cattle. Antibody responses of cattle against major surface proteins (MSP) 1a, 1b and 5, as determined by ELISAs, peaked 2 weeks after the last immunization. Cattle immunized with infected IDE8 cell-derived antigens had a preferential recognition for MSP1b while cattle immunized with erythrocyte-derived antigens had a preferential recognition for MSP1a. Protection efficacy was evaluated using the percent infected erythrocytes (PPE), the packed cell volume (PCV), and the prepatent period. A. marginale-immunized cattle showed lower PPE and higher PCV values when compared to control animals and did not display clinical anaplasmosis. The cell culture-derived A. marginale shows promise for use as antigen in development of a new killed vaccine for anaplasmosis.

Anaplasma↗

DNA binding properties of the ecdysteroid receptor in the salivary gland of the female ixodid tick, Amblyomma hebraeum.

Salivary gland degeneration in the female tick, Amblyomma hebraeum Koch (Acari: Ixodidae) is controlled by an ecdysteroid hormone. In an earlier study (Mao, H., McBlain, W.A., Kaufman, W.R., 1995. Some properties of the ecdysteroid receptor in the salivary gland of the ixodid tick, Amblyomma hebraeum. Gen. Comp. Endocrinol. 99, 340-348), we demonstrated that a protein component of a salivary gland extract binds to ponasterone A (Pon A) with high affinity (Kd-1 nM), suggesting a tick ecdysteroid receptor (EcR). In this study, the Pon A binding protein bound to calf thymus DNA; this binding could be dissociated by Drosophila hsp27 EcRE. The binding protein shifted the [32P]hsp27 EcRE band on a gel mobility shift assay; formation of the complex with hsp27 EcRE required KCl (optimal concentration was approximately 75 mM). A number of physiologically effective ecdysteroids enhanced the binding with the following order of potency: Pon A > Mur A > Mak A > 20E > ecdysone, whereas vertebrate steroids (estradiol, cholesterol, corticosterone, progesterone, testosterone) had no such effect. Using monoclonal antibodies against Drosophila EcR and USP, we found that AG 10.2 recognized three bands (90.5, 87.3 and 84 kDa for EcR) and AB11 recognized at least two major bands (50.3 and 47.1 kDa for USP) in the salivary gland extract by western blot analysis. In addition, AB11 supershifted the tick EcR-hsp27 EcRE band on a gel mobility shift assay, indicating that the tick EcR heterodimerized with a USP-like protein for DNA binding. Furthermore, selective mutations to the 15-basepair palindrome of hsp27 EcRE at positions-5, + 2, or adding a base to the spacer, resulted in considerably reduced affinity to the tick EcR/USP. We thus propose a sequence similarity of EcREs between A. hebraeum and its insect counterpart.

Animals↗

Clinical and epidemiological features of early Lyme disease and human granulocytic ehrlichiosis in Wisconsin.

To compare clinical features and assess risk factors for human granulocytic ehrlichiosis (HGE) and early Lyme disease, we enrolled patients in a case-control study during the 1996 and 1997 tick seasons. Clinical and demographic characteristics were assessed for patients with laboratory-confirmed cases of HGE or Lyme disease, and risk factors were compared with those of matched control subjects. We identified 83 persons with Lyme disease, 27 with HGE, and 11 with apparent coinfection. Unsuspected Ehrlichia infection was identified in 8 (13%) of 60 patients with Lyme disease. Patients with HGE were older and more likely to have fever, chills, or dyspnea than were those with Lyme disease only. Most patients with apparent coinfection did not have hematologic abnormalities. In the risk factor analysis, tickborne illness was independently associated with rural residence and camping. The clinical spectrum of HGE overlaps that of Lyme disease, and physicians in areas of endemicity should consider both diseases in treating patients with a compatible rash or febrile illness.

Adult↗

Effect of cyclophosphamide on the acquisition of resistance to infestation by Rhipicephalus appendiculatus in rabbits.

Parenteral administration of cyclophosphamide in rabbits made resistant to infestation by the tick Rhipicephalus appendiculatus, resulted in abrogation of the resistance phenomenon. As a result, a high percentage (83%) of the larval ticks fed to repletion. This was in contrast to the control rabbits which were expressing the phenomenon of resistance to infestation by the ticks. In the latter, only 28% of the ticks fed to repletion. Cyclophosphamide administration in rabbits induced a leucopenia and depressed antibody response to the heterologous antigen, sheep red-blood cells.

Animals↗

Regeneration of Haller's sensory organ in the tick Ixodes ricinus L.

Amputation of legs in nymphs of ticks, obtained from the first laboratory generation, resulted in regeneration of the legs after moulting to adults. Haller's sensory organ on the upper surface of each foreleg tarsus was significantly modified following regeneration. Haller's organ in non-amputated legs of the experimental ticks remained unchanged, being comparable to controls. Pored olfactory sensilla in the anterior pit, in a capsule and on a distal knoll usually increased in number, as well as grooved, thin and conical sensilla. Bordering gustatory and double-walled postcapsular sensilla either decreased or increased in number. All additional sensilla were consistent in their location. Form of the anterior pit and capsule's aperture also deeply changed after the regeneration. The authors distinguish this changes as atavistic. No correlation between changes in different parts of the organ were found. A phenomenon of induction was discovered in our study: if a distal part of tick's gnathosoma was amputated together with the left foreleg, Haller's organ in the right, untreated leg possessed the same changes after moulting as the regenerated organ in the previously amputated left foreleg.

Animals↗

[The entomological and acarological situation in Nizhegorod Province].

Ixodes, persulcatus, I. ricinus, D. reticulatus, D. marginatus, H. concinna were recorded in different regional environmental regions. The northern districts of the region are endemic for tick-borne encephalitis (TBE) and Ixodes tick-borne borreliosis (ITBB). The infection of ticks with TBE virus is 0.25%, that with Borrelia is 10.3% of the total number of the examinees. In 1997 TBE and TTBB morbidity rates were 1.91 and 2.0 per 100,000, respectively. Data on the phenology of prevailing Anopheles messeae are given. Human malaria contamination may take 99-102 days (the first ten days of June to the second ten days of September). The large size of malaria mosquitoes and the presence of imported cases of malaria make it possible to deteriorate the malaria epidemiological situation. The larvicidal activity of a preparation derived from shepherd's-purse (Capsella bursa pastoris) [correction of caseweed (Bursae pastoris)] was tested. 80-85% deaths of larvae of second-third ages occurred 48 hours after treatment with the preparation, 1 kg/ha.

Animals↗

Effects of gamma radiation on spermatogenesis and fertility of male Amblyomma americanum (Acari: Ixodidae).

Amblyomma americanum males were treated with 0.5, 1, 2, 3, 4, 8, and 16 krad of gamma radiation. Testes of ticks treated with 2, 3, 4, 8, and 16 krad were smaller than those of ticks irradiated at lower levels and controls. No recognizable alteration in timing of spermatogenesis was noted among the different radiation groups, but severe breakdown and depletion of germinal cells was noted at 4, 8, and 16 krad. Percent hatch of larvae from crosses of irradiated males and untreated females decreased with increasing radiation level. No hatch was observed from eggs of females mated to males treated at 2 krad or higher.

Animals↗

Vertical movement and posture of blacklegged tick (Acari: Ixodidae) nymphs as a function of temperature and relative humidity in laboratory experiments.

We measured three components of the behavioral activity of blacklegged tick (Ixodes scapularis Say) nymphs under controlled conditions of daylength, temperature, and relative humidity in the laboratory. Temperature and relative humidity were experimentally manipulated among replicated treatment groups in two experiments. Humidity treatment had a significant effect on mean questing height, which was greater at 100% RH than at any lower humidity (P < 0.01), but it had no effect on mean distance moved between observations or the mean percentage of time in questing posture. Temperature had no effect on mean questing height, but significantly affected mean distance moved (P < 0.01) and the percentage of time in questing posture (P < 0.001). Mean distance moved and percentage of time in questing posture were both greater at 25 degrees C than at higher or lower temperatures. A mechanistic understanding of the factors affecting tick behavior is likely to be useful for interpreting field data, designing field studies, and predicting risk of exposure to tick-borne pathogens.

Animals↗

Suppression of subadult Ixodes scapularis (Acari: Ixodidae) following removal of leaf litter.

Removal of leaf litter in wooded areas of a forested residential community significantly reduced the abundance of questing blacklegged tick, Ixodes scapularis Say, nymphs throughout their peak activity period. Removal of leaf litter, using hand rakes and leaf blowers during the early spring (March) and early summer (June) resulted in reductions in nymphal tick density ranging from 72.7 to 100%. Subsequent sampling of the I. scapularis population during the summer revealed similar rates of suppression of larvae. Although leaf removal provides levels of suppression similar to more traditional methods, including chemical acaricides, it is labor intensive and not free of possible nontarget effects. The potential role of leaf litter removal in an integrated program to control I. scapularis is discussed.

Animals↗

Effects of cattle tick (Boophilus microplus) infestation on the bovine immune system.

The immunosuppressive effect of experimental Boophilus microplus infestation on bovine peripheral blood lymphocytes (PBL) and on host antibody production to a protein antigen (ovalbumin) was examined. Boophilus microplus infestation caused a marginal decrease in the percentage of T lymphocytes in PBL, which was observed in both lightly (5000 larvae) and heavily (40,000 larvae) infested cattle, and began at the second infestation and continued until the end of the fourth infestation. The percentage of B lymphocytes in heavily tick-infested cattle was less than that in non-infested control cattle after the fourth infestation. The response of PBL from tick-infested cattle to phytohemagglutinin (PHA) was always less than that of tick-free cattle after the second infestation. No noteworthy differences were detected between the three stages of tick infestation, that is, 1 week before the peak of adult engorgement, the middle of the peak and 1 week after all ticks had dropped. Boophilus microplus saliva (100 microliters ml-1) suppressed 47% of the response of bovine PBL to PHA in vitro. This suppressive effect of saliva may contribute to the lower responsiveness of PBL from tick-infested cattle. Antibody production by tick-infested cattle was examined during the third and fourth heavy tick infestation. Tick-infested cattle showed a diminished response against ovalbumin after the second immunization. The immunosuppressive effects of tick infestation may play an important role in tick survival or in the transmission of tick-borne diseases in the field.

Animals↗

Effects on cattle of artificial infestations with the tick Rhipicephalus appendiculatus.

Groups of ten cattle were exposed to 0, 40 and 400 adult Rhipicephalus appendiculatus feeding once a week for 24 weeks. Although no differences in cumulative percentage weight gain were found at the end of the experiment, animals free of ticks performed better during the first 12 weeks. A marked recovery of the 400-tick group was recorded during weeks 13-24 and after ticks were removed. Haemoglobin and packed cell volume values were lowered in tick-infested animals but no clear trends were observed in red and white blood cells. No differences between the percentages of ticks feeding in the 2 tick-infested groups on day 2 were observed but higher numbers of females were feeding on the 40-tick group on day 5. More ticks were found on day 5 after tick application to the cattle during weeks 1-6 than in the rest of the experiment. Significant correlations were found between lymph node enlargement and ear damage when they were tested against tick load. R. appendiculatus nymphs feeding on the formerly tick-infested cattle were fewer, lighter and engorged in a shorter time than those on the control animals. The possible causes for the reduction in ticks feeding, the changes in cattle weights, blood composition and development of host resistance are discussed.

Analysis of Variance↗

Epidemiologic evaluation of the risk factors associated with exposure and seroreactivity to Bartonella vinsonii in dogs.

OBJECTIVES: To determine seroprevalence to Bartonella vinsonii subsp berkhoffii in a population of sick dogs from North Carolina and Virginia and to evaluate potential risk factors associated with increased likelihood of exposure to the organism. SAMPLE POPULATION: Serum samples from 1,920 sick dogs. PROCEDURE: An indirect fluorescent antibody assay was performed on each sample, and the end-point antibody titer was recorded. A case (seropositive) was defined as a dog with reciprocal titer > or = 64, and a control (seronegative) was defined as a dog with reciprocal titer < 16 that was referred within 0 to 3 days of referral of a corresponding case. From this population, 207 dogs (69 cases and 138 controls) were included in a case-control seroepidemiologic study. RESULTS: 3.6% (69/1,920) of the dogs were seropositive to B vinsonii subsp berkhoffii. Results of the case-control study indicated that seropositive dogs were more likely to live in rural environments, frequently on a farm, were free to roam the neighborhood, and were considered to be predominantly outdoor dogs. Moreover, seropositive dogs were 14 times more likely to have a history of heavy tick exposure. After analysis of the case-control study, a more detailed examination of banked sera from dogs with known tick exposure was performed. High correlation was found between sero-reactivity to B vinsonii and seroreactivity to E canis or B canis (36.0 and 57.1%, respectively). Sera derived from dogs experimentally infected with E canis or R rickettsii did not cross react with B vinsonii antigen. CONCLUSIONS AND CLINICAL RELEVANCE: Several potential risk factors are associated with canine exposure to B vinsonii. Rhipicephalus sanguineus, the tick vector for E canis and B canis, may be involved in B vinsonii transmission among dogs.

Animals↗

Distribution of some anystid mites (Acari: Anystidae) in Australia and Indonesia and their role as possible predators of the cattle tick, Boophilus microplus (Acari: Ixodidae).

The geographic and habitat distributions of Anystis baccarum, A. jabanica, A. salicinus, and A. sp. B Walzia australica Womersley and Chaussieria warregense in Australia and parts of Indonesia are presented. Laboratory trials showed that A. baccarum will feed upon the larvae of the cattle tick Boophilus microplus, whilst A. jabanica also showed some predatory potential. Walzia australica and C. warregense had no effect on survival of tick larvae.

Animals↗

Exposure of cattle immunized with different stocks of Theileria parva to buffalo-associated Theileria challenge on two game parks in Zimbabwe.

Eight cattle immunized with cattle-derived Theileria parva Boleni stabilate together with six susceptible controls were released in Dombawera Game Park on the Highveld of Zimbabwe. This coincided with Rhipicephalus appendiculatus nymphal activity. The cattle grazed together with African buffaloes (Syncerus caffer) and were not treated against tick infestation. The nymphal tick infestation was high, and seven of the eight immunized cattle and three of the controls had severe and fatal reactions. Subsequently, two stocks of Theileria parva to be tested for their immunizing abilities were prepared-one from adult ticks which were fed as nymphs on one of the sick control animals (Dom 268) and the other from adult ticks collected from pastures grazed by buffaloes (Bv-1). Two groups of cattle were immunized with either the Dom 268-derived strain (eight animals) or the Bv-1-derived strain (four animals). These together with three non-immunized controls, were released in Bally Vaughaun Game Park in the Highveld, where buffaloes are present, during the season of nymphal tick activity. A third group of five cattle, immunized with stabilate Bv-1, and three non-immunized controls were released at the same site during the season of adult tick activity. The nymphal and adult tick infestations of the cattle were large and more than 2000 nymphs and 1000 adult ticks were counted per animal. Cattle were treated with a pyrethroid pour-on preparation to control the tick infestation and screw-worm strike. The immunized cattle in the three groups survived the theileriosis challenge for a period of 18 months, but the non-immunized control cattle suffered a severe and fatal theileriosis 19-23 days after being placed on the pasture.

Animals↗