Immunity against female Ixodes ricinus L.: effect on feeding and haemoglobin digestion

Rabbits have been infested 3 times with 10 females and 10 males Ixodes ricinus. Immunity which is acquired when ticks feed on naive rabbits (first infestation) perturbs tick feeding on reinfested animals (third infestation). Then ticks ingest less blood (p less than 0.001). Blood meal digestion in also altered. It was estimated by measuring haemoglobin concentration in ixodid midgut during 20 days after their drop off. After the first infestation this concentration decreases linearly with time (r2(1) = 46.14%, n1 = 63, p less than 0.001). After 3 infestations it is no longer correlated with time, indicating an impaired digestive process (r2(3) = 7.15%, n3 = 49, p greater than 0.05). This observation was corroborated by an analysis of multiple regression. Haemoglobin concentration of tick midgut only correlates with time after a first infestation (r2(1) = 45.25%). In ticks fed on immune animals this concentration is predicted with the quantity of midgut C3 and the weight of fed ticks and not with time (r2(3) = 60.99%).     

Performance of female Rhipicephalus sanguineus (Acari: Ixodidae) fed on dogs exposed to multiple infestations or immunization with tick salivary gland or midgut tissues

This investigation compared the effects of repeated infestations to immunization of dogs with tick salivary gland or midgut extracts on the feeding and fecundity performances of female Rhipicephalus sanguineus (Latrielle). In each immunized group, three tick-naive dogs were immunized three times with tick salivary gland or midgut extracts, and twice challenged at 21-d intervals by allowing 80 female and 40 male adult ticks to feed on each host. The repeated infestation group of three naive dogs was infested five times at 21-d intervals by the same numbers of ticks. The repeated infestation group showed a trend of reduced tick performance after the third infestation, but some of the tick performance parameters had recovered by the fifth infestation. Tick attachment was reduced by immunization with either tick salivary gland or midgut extract. Immunization with tick salivary gland extract had the greatest impact on the feeding period and engorgement weight of the female ticks. Immunization with tick midgut extract resulted in the greatest reduction of tick fecundity parameters, which included preoviposition, oviposition, and egg-incubation periods in addition to reduced egg production and egg viability. These results confirm that dogs can become resistant to R. sanguineus, and demonstrate that immunization with tick salivary gland or midgut extract has different effects on tick feeding and fecundity.     

Ticks (Acari: Ixodidae) on wild marsh-deer (Blastocerus dichotomus) from Southeast Brazil: infestations before and after habitat loss

The lake from Porto-Primavera hydroelectric power station inundated an area of 2,200 km2 at the border of S?o Paulo and Mato-Grosso do Sul States, Brazil. Infestations by ticks were evaluated on 135 marsh deer, Blastocerus dichotomus (Illiger), captured before and after inundation. Ticks were collected for identification, and infestation level of animals was assessed by scoring. Deer were divided into four groups according to capture location and temporal relation to the inundation. Groups 1, 2, and 3 were captured before inundation. Group 4 was captured after inundation. Four tick species were found: Amblyomma cajennense (F.), Amblyomma triste Koch, Anocentor nitens (Neumann), and Boophilus microplus (Canestrini). Groups 1, 2, 3, and 4 had 30, 45, 100, and 96%, respectively, of animals carrying B. microplus ticks. A. triste was observed on 16, 22, 22, and 88% of animals from groups 1, 2, 3, and 4, respectively. A. nitens and A. cajennense were observed only on group 4, on 32 and 16% of the animals, respectively. Groups 1 and 2 had only 4.8 and 6.1% of animals with high infestation levels, respectively, and no ticks on 46.8% and 45.5% of the animals, respectively. Conversely, groups 3 and 4 lacked noninfested animals and had high infestation levels on 77.8 and 50% of deer, respectively. Marsh area shrinkage was blamed for higher infestation levels on deer from groups 3 and 4. The widespread presence of A. triste on marsh deer, a Neotropical tick species, raises the possibility of a natural host-parasite relationship.     

Attempt to control ticks (Acari: Ixodidae) on deer on an isolated island using ivermectin-treated corn

We report an attempt by an offshore island community to control the vector tick of Lyme disease by providing ivermectin-treated corn to an isolated herd of free-ranging white-tailed deer, Odocoileus virginianus Zimmerman. Medicated corn was supplied in troughs within the island village and from automatic feeders at remote sites during 5 consecutive fall and spring adult tick questing seasons. Acaricide consumption was monitored by assaying its presence in fresh deer pellets and its concentration in deer sera. Its effectiveness was evaluated by recording the number of adult ticks collected from the hides of deer, the number of females becoming sufficiently engorged to oviposit, and the success of subsequent oviposition and eclosion. Entomologic risk was monitored by collecting immature ticks from hosts and adult ticks from vegetation. Estimates based on a subsequent deer reduction program indicated that up to twice as many deer had been present during the project as originally presumed. For this and other reasons related to deer behavior, target levels of serum ivermectin were achieved in a minority of deer. Nevertheless, > 90% control of female tick infestation, subsequent oviposition, and larval eclosion was obtained in those 8 of 16 sampled deer with serum ivermectin levels of > or = 15 ng/ml. In addition, the ratio of females to males, the numbers of females engorging > 10 mg body weight, and the numbers of those eventually hatching, were all significantly less among ticks from island deer in comparison with ticks from untreated deer. No consistent changes in the numbers of ticks found on immature-stage hosts or removed from vegetation were noted within 3 yr of the cessation of treatment.     

Quantitative description of the development of Babesia ovis in Rhipicephalus bursa (hemolymph,ovary, eggs)

The development and infection dynamics of Babesia ovis in the hemolymph, ovaries, and eggs of Rhipicephalus bursa are described quantitatively, based mainly on examination of Giemsa-stained smears. After alimentary infection of female ticks, their hemolymph became infected 5 days after repletion (p.repl.). The prevalence and mean intensity of infection increased during the course of infection studied, up to 17 days p.repl. After vertical infection of female ticks, their hemolymph was infected only during the first 3 days after the onset of infestation (p. infest.) and again after the onset of alimentary infection 5 days p.repl. There was a positive correlation between prevalence and mean intensity of infection in the hemolymph. The prevalence of infection decreased with aging of the unfed adult ticks. After alimentary infection, the ovaries became infected 6 days p.repl., and after vertical infection, 3 days p. infest; they remained infected until the death of the tick. Ticks selected for susceptibility during 18 and 19 vertically infected generations were more susceptible than ticks in their first to third vertically infected generations or alimentarily infected ticks. Eggs deposited on day 1 of oviposition were noninfected after alimentary infection of the female tick. After vertical infection of the tick, even such eggs became infected; the infection, then, was detectable in eggs produced throughout the oviposition period regardless of the infection mode. Intense hemolymph infections induced an increase of egg degeneration and a decrease of total as well as infected egg production. There was a positive correlation between the number of deposited and infected eggs as well as between prevalence and mean intensity of infection in eggs.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunization of guinea-pigs against Rhipicephalus appendiculatus adult ticks using homogenates from unfed immature ticks.

Guinea-pigs immunized with homogenates of unfed larvae and nymphs of the tick Rhipicephalus appendiculatus developed significant levels of protective immunity to infestation with adults of this species. The mean engorged weight of female ticks feeding on immunized animals (181.96 +/- 05.63 mg and 170.11 +/- 11.54 mg) was reduced by an average of 46% and 51%, respectively, compared to that of female ticks feeding on control guinea-pigs, although in some individual animals the reduction was as high as 86%; the mean egg mass weight was also significantly reduced. Electrophoretic separation of the homogenates followed by immunostaining with post-infestation sera revealed several antigen bands common to all stages. Two bands of 36,500 and 23,000 molecular weight (MW) were recognized in all homogenates by post-adult infestation serum, but not by post-larval or post-nymphal infestation sera, suggesting that these may be antigens specifically involved in feeding by adult ticks, and are either not presented to the host's immune system or presented only in minimal amounts during feeding by immature stages. Sera from animals immunized with the homogenates did not recognize either of these antigens. Post-immunization sera did, however, stain two bands of 84,000 and 60,000 MW in the homogenates which were not recognized by post-infestation sera.     

Cyclosporin A prevents suppression of delayed-type hypersensitivity in mice immunized with high-dose sheep erythrocytes.

When administered intraperitoneally to mice 2 days before immunization with a tolerogenic dose (10(9)) of sheep red blood cells (SRBC), cyclosporin A (CsA; 200 mg/kg) strikingly augmented 4-day delayed-type hypersensitivity (DTH) footpad reactions. These enhanced responses were similar in magnitude to those seen in animals sensitized with an immunogenic, low-dose (10(6)) SRBC. The stimulatory effect of CsA was observed over the dose range of 5-200 mg/kg and was obtained in animals given the drug in one injection, up to 7 days before sensitization. The augmentation of DTH was characterized by footpad swelling, intense mononuclear cell infiltration and increased deposition of 125I-fibrinogen within the challenge site. In addition, increased expression of procoagulant activity by spleen cells in response to antigen was observed. Cell transfer experiments showed that the CsA-enhanced DTH could be adoptively transferred to naive recipients. Additional transfers conducted at the time of antigen challenge suggested that, under the conditions described, CsA inhibited the action of a population of suppressor cells normally effective during DTH reactions.     

Tick infestation on wild snakes in northern part of western Ghats of India

In total, 167 individuals of 30 species of snakes belonging to 22 genera and five families were examined for tick infestation from November 2008 to March 2010. Only two species of snakes, Ptyas mucosa (L., 1758) (Indian rat snake) and Naja naja (L., 1758) (spectacled cobra), were found infested by ticks. All ticks collected were identified to be Amblyomma gervaisi [previously Aponomma gervaisi (Lucas, 1847) 1. The average prevalence of these ticks on Indian rat snakes (n=48) was 29.16%, with abundance of 7.02 ticks per individual; on spectacled cobras (n=20), average prevalence was 30.00%, with abundance of 6.9 ticks per individual. The nymphs and males were predominant. All the ticks were found on the dorsal aspect of the body of the snake, and no ticks were recorded on the head, tail, or ventral body. The rate of tick infestation was highest in scrubland and was lowest in evergreen forests. Female Indian rat snakes showed higher tick infestation rates than male Indian rat snakes. Using Mann-Whitney U test, we found that longer snakes of both species had significantly higher rate of tick infestation in both the species of snakes.     

Mechanisms of eosinophilia in mice infested with larval Haemaphysalis longicornis ticks.

Infestation of larval Haemaphysalis longicornis ticks induced a threefold increase of eosinophils in the peripheral blood of normal WBB6F1- +/+ mice 2 days after tick infestation. In genetically mast cell-deficient WBB6F1- W/Wv mice, a threefold increase of blood eosinophils was observed 6 days after the tick infestation. However, marked infiltration of eosinophils was detected in the tick infestation sites of the WBB6F1- +/+ mice but not the WBB6F1- W/Wv mice. When the mast cell deficiency of WBB6F1- W/Wv mice had been rescued locally by intradermal injections of WBB6F1- +/+ mouse-derived cultured mast cells, a rapid increase of blood eosinophils and tissue infiltration of eosinophils were revealed following tick infestation. The intravenous (i.v.) injection of immune spleen or lymph node cells obtained from WBB6F1- +/+ mice 10 days after tick infestation led to significant eosinophilia in naive recipient mice. Treatment with anti-Thy-1.2 or anti-CD4 monoclonal antibody (mAb) and complement (C) completely abolished the eosinophilia; the early response (2 days after tick challenge) is dependent on mast cells at the feeding site, and the late response (6 days after tick challenge) is dependent on T lymphocytes. Since amplified interleukin-5 (IL-5) cDNA was detectable in the spleen cells 4 days after tick infestation, the late response might be mediated by IL-5. The infiltration of eosinophils at the feeding site of skin appeared to be dependent on mast cells.     

Ability of the vector tick Boophilus microplus to acquire and transmit Babesia equi following feeding on chronically infected horses with low-level parasitemia

The protozoan parasite Babesia equi replicates within erythrocytes. During the acute phase of infection, B. equi can reach high levels of parasitemia, resulting in a hemolytic crisis. Horses that recover from the acute phase of the disease remain chronically infected. Subsequent transmission is dependent upon the ability of vector ticks to acquire B. equi and, following development and replication, establishment of B. equi in the salivary glands. Although restriction of the movement of chronically infected horses with B. equi is based on the presumption that ticks can acquire and transmit the parasite at low levels of long-term infection, parasitemia levels during the chronic phase of infection have never been quantified, nor has transmission been demonstrated. To address these epidemiologically significant questions, we established long-term B. equi infections (>1 year), measured parasitemia levels over time, and tested whether nymphal Boophilus microplus ticks could acquire and, after molting to the adult stage, transmit B. equi to naive horses. B. equi levels during the chronic phase of infection ranged from 10(3.3) to 10(6.0)/ml of blood, with fluctuation over time within individual horses. B. microplus ticks fed on chronically infected horses with mean parasite levels of 10(5.5) +/- 10(0.48)/ml of blood acquired B. equi, with detection of B. equi in the salivary glands of 7 to 50% of fed ticks, a range encompassing the percentage of positive ticks that had been identically fed on a horse in the acute phase of infection with high parasitemia levels. Ticks that acquired B. equi from chronically infected horses, as well as those fed during the acute phase of infection, successfully transmitted the parasite to naive horses. The results unequivocally demonstrated that chronically infected horses with low-level parasitemia are competent mammalian reservoirs for tick transmission of B. equi.     

Acquired resistance to ticks. I. Passive transfer of resistance.

Guinea-pigs developed resistance to larvae of the ixodid tick, Dermacentor andersoni, after one infestation. Resistance was characterized by guinea-pigs allowing fewer larvae to engorge (5-15%) during a second exposure than during an initial infestation (70-90%). Larvae feeding on resistant hosts weighed less than larvae engorging on a host with no previous exposure to ticks. Evidence is presented which indicates that this resistance can be passively transferred with viable lymph node cells, but not with serum, from resistant guinea-pigs. Recipients of cells from such resistant animals allowed significantly fewer larvae to engorge than did controls previously unexposed to ticks. This was not so in recipients of immune serum. The protection provided by passive transfer of cells from a resistant donor was not as complete as the protection afforded by natural exposure.     

Feeding efficiency of larval Hyalomma truncatum (Acari: Ixodidae) on hosts previously exposed to ticks.

The effect of guinea pig response to feeding larval Hyalomma truncatum Koch ticks was studied by measuring the percentage of ticks engorging and molting, their engorged weight, and hemoglobin content. Four guinea pigs were infested with 100, 200, 400, and 1,000 larvae, respectively, on three occasions at 21-d intervals, followed by a fourth infestation with 500 larvae. During the second, third, and fourth infestations, significantly fewer ticks engorged on each guinea pig than during the first infestation. The greatest reduction in percentage molting occurred during the fourth infestation on the animal with successive exposure to 400 larvae; only 24% of the ticks that fed molted. Ticks with the lowest mean weight and hemoglobin content also engorged on this animal during the fourth infestation. Guinea pigs exposed to 200 and 400 H. truncatum larvae elicited the greatest change in feeding efficiency during the fourth infestation. However, these hosts had no effect on a single subsequent fifth infestation with Amblyomma cajennense (F.) nymphs, as greater than 95% successfully engorged.     

Histopathology and Ultrastructure Features of the Midgut of Adult Females of the Tick Amblyomma cajennense Fabricius, 1787 (Acari: Ixodidae) in Various Feeding Stages and Submitted to Three Infestations

ABSTRACT

The digestive tube of the tick Amblyomma cajennense is responsible for the digestion during feeding on the host. This study analyzed the midgut of unfed, partially engorged, and fully engorged fed females as well as three infestations in rabbits. In A. cajennense, the digestive tube is long and from the midgut, two pairs of diverticula ramify and lead to a blind end. In some midgut regions were observed for the first time in ticks, structures termed here “nodules.” The midgut of unfed females possesses a pseudostratified epithelium composed of digestive and generative cells. In partially engorged and engorged females at 1st infestation and partially engorged at 2nd infestation, the epithelium becomes stratified. In partially engorged females at 2nd infestation, the epithelium exhibits a third cell type: secretory cell. So the intestinal epithelium undergoes several changes during the feeding process in ticks at subsequent infestations. As infestations progress in the same host, the latter becomes more resistant and female ticks require more days to complete their feeding cycle, which in A. cajennense is 25 days.     

Cutaneous basophil-associated resistance to ectoparasites (ticks). I. Transfer with immune serum or immune cells.

Immune resistance experiments were carried out in guinea-pigs employing two tick species that as adults are ectoparasites of cattle (Ixodes holocyclus and Rhipicephalus appendiculatus). These studies showed that susceptibility of non-immune guinea-pigs to infestation with tick larvae varies according to the species of tick and the strain of guinea-pig. With both tick species, greater than 90% acquired resistance was achieved in several guinea-pig strains. Immune resistance was evident within a week following primary infestation and lasted up to 9 months following a single sensitizing exposure to tick feeding. The strength and duration of resistance was influenced strongly by the size of the initial sensitizing dose. Immune resistance was readily transferred to naive recipients by intravenous administration of either peritoneal exudate cells or immune serum from donors sensitized by a single prior infestation with ticks. Doses of serum as small as 0.5 ml transferred resistance. These studies demonstrate that both sensitized cells and immune serum factors contribute significantly to acquired host resistance to ticks that as adults are ectoparasites of cattle.     

Efficacy of a novel fipronil spot-on for the treatment and control of induced infestations of adult cat fleas (Ctenocephalides felis) and castor bean ticks (Ixodes ricinus) on cats

Cat fleas (Ctenocephalides felis) and the castor bean tick (Ixodes ricinus) cause discomfort and health effects due to bites and ingestion of blood and they serve as vectors for several animal and human pathogens. Effectiveness of a novel 10 % w/v fipronil spot-on (Eliminall®/Exproline vet?, marketed by Pfizer Animal Health and registered and manufactured by Krka, d.d., Novo mesto) was confirmed against these parasites on experimentally infested cats. Two parallel, unicentre and masked controlled studies were conducted with European mixed breed and mixed sex cats. Cats were allocated randomly to one of two treatment groups based on either pre-treatment flea counts (study 1) or pre-treatment tick counts (study 2). In each of the study, eight animals served as control, while another eight animals were treated once topically with the unit label dose of 50 mg fipronil per cat (10.6–23.8 mg/kg). At each reinfestation, animals were infested with approximately 100 fleas or 60 ticks to achieve adequate infestation rates. Parasites were removed and counted on days 2, 9, 16, 23, 30 and 37, 48 h after the treatment or experimental infestation. Excellent effectiveness was demonstrated on day 2 (100 and 94 % efficacy against fleas and ticks, respectively) and lasted for up to 5 weeks (efficacy ≥96 %) against fleas and up to 4 weeks against ticks (efficacy ≥94 %). The product was well tolerated and no adverse reactions were observed.     

Reservoir role of lizard Psammodromus algirus in transmission cycle of Borrelia burgdorferi sensu lato (Spirochaetaceae) in Tunisia

To investigate the reservoir role of the lizard Psammodromus algirus for the Lyme disease spirochete, 199 lizards were trapped from April to October 2003 in El Jouza, northwestern Tunisia. In this site, the infection rate of free-living Ixodes ricinus (L.) by Borrelia was evaluated by immunofluorescence as 34.6% for adult ticks and 12.5% for nymphs. Eighty percent of P. algirus (117/146) captured during this study were infested by I. ricinus, the predominant tick species collected from lizards. The intensity of tick infestation of this host by larvae and nymphs ranged from 0.14 to 7.07 and from 1.5 to 6.58, respectively. These immature stages of I. ricinus were found on lizards in spring and the beginning of summer, with a peak of intensity during June (10.16 immature ticks by lizard). Tissue cultures from lizards and xenodiagnosis with larval I. ricinus were used to assess the infection and the ability, respectively, of infected lizards to transmit Borrelia to naive ticks. Seventeen percent of xenodiagnostic ticks (40/229) acquired B. lusitaniae while feeding on P. algirus. Therefore, we demonstrated the ability of the lizards to sustain Borrelia infection and to infect attached ticks, and we proved that P. algirus is a reservoir host competent to transmit B. lusitaniae.     

Glass capillary tube feeding: a method for infecting nymphal Ixodes scapularis (Acari: Ixodidae) with the lyme disease spirochete Borrelia burgdorferi

We evaluated an artificial capillary feeding method to infect nymphal Ixodes scapularis (Say) ticks with Borrelia burgdoeferi, the causative agent of Lyme disease. Thirty to 70% of the nymphs were infected after feeding for 2.5 h from glass capillary tubes filled with a solution of spirochetes. Capillary infection was stable and persisted in the nymphs for at least 10 d after feeding. Capillary feeding also maintained natural vector competence patterns because I. scapularis ticks acquired infection unlike Dermacentor variablis (Say), which did not become infected. Capillary infected I. scapularis nymphs were capable of transmitting the infection to naive mice although not as efficiently as naturally infected nymphs. The capillary infection method is convenient and is a better alternative to syringe inoculation as a means of infecting animals with B. burgdorferi.     

Changes in the concentration of globulins in naive guinea pigs during feeding by the immature stages of Rhipicephalus evertsi evertsi (Acari: Ixodidae)

The weight of Rhipicephalus evertsi evertsi Neumann nymphs, which as larvae and nymphs completed the entire blood meal on the same individual guinea pig, was significantly lower than the weight of those that as larvae and nymphs fed on two separate naive hosts. Nymphs of the latter category spent 1 wk (from unfed larvae to unfed nymphs) on one animal before their transfer to the second host to complete the blood meal. The albumin concentration of the host blood decreased and was related to the time that the immature ticks spent on the hosts. The albumin/globulin ratio also decreased. The alpha 1 globulin fraction increased soon after the guinea pigs were infested with ticks. No further changes in the levels of the alpha 1 globulin fraction were observed with time. The concentration of serum beta globulins increased only in guinea pigs infested with immature ticks for the entire larval and nymphal feeding period. A second infestation of those guinea pigs with larvae of R. e. evertsi resulted in further increases in the levels of serum beta globulins. The relationships among serum beta globulin levels, the weight of engorged nymphs, and host immunity are discussed.     

Immunomodulatory effects of cyclosporin A on human peripheral blood dendritic cell subsets

Cyclosporin A (CsA) is a potent immuno-suppressant and is approved for the treatment of various disease conditions. The molecular biological mechanism of CsA has been investigated intensively in T cells and has been shown to involve the intracellular calcineurin pathway. Recently, it was reported that CsA has capacities to affect not only T cells but also antigen-presenting cells such as B cells and dendritic cells (DCs). DCs are a master regulator of immune responses that have an integral capacity to prime naive T cells. In the present study, we investigated the biological effects of CsA on human peripheral blood DC subsets: CD11c+ myeloid and CD11c- lymphoid subsets. CsA inhibited the up-regulation of co-stimulatory molecules induced with or without microbial stimuli and CD40L on both CD11c+ and CD11c- subsets. In addition, CsA negatively regulated the endocytic activity of CD11c+ DC during the immature state. CsA inhibited the interleukin-12 (IL-12) production, but augmented the IL-10 production from the LPS-stimulated CD11c+ subset, whereas CsA reduced the interferon-alpha (IFN-alpha) production from the CD11c- subset infected with Sendai virus (SV). Both the LPS-stimulated CD11c+ subset and SV-infected CD11c- subset preferentially induced the development of IFN-gamma-producing T helper-type 1 (Th1) cells. Pretreatment of these DC subsets with CsA inhibited the Th1 skewing. These findings suggested a DC-mediated mechanism of immunosuppression by CsA.