The possible use of integumentary sense organs as specific diagnostic mean for larvae and nymphs of Rhipicephalus sanguineus and Rhipicephalus turanicus.
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Relationships between species of the Rhipicephalus sanguineus group: a molecular approach.
The biosystematic status of species belonging to the Rhipicephalus sanguineus group is difficult to determine by phenotypic methods and has been subject to frequent revisions and ongoing debate. Yet, the differentiation of species within this complex is of clinical importance especially in veterinary medicine due to different host specificities of the transmitted pathogens. In order to generate quantifiable and comparable data independent of environmental influences, a section of the second internal transcribed spacer (ITS2) of the rRNA gene was sequenced for 6 species belonging to the Rhipicephalus sanguineus complex: Rhipicephalus sanguineus sensu stricto, R. turanicus, R. rossicus, R. pumilio, R. pusillus, and R. camicasi. Rhipicephalus sanguineus s. str. and R. turanicus as well as R. pumilio and R. rossicus were found to have a close genetic relationship compatible with possible conspecificities. Conversely, R. pusillus was found to have the most distant relationship to the other species.
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Abnormal development of Rhipicephalus sanguineus (Ixodidae).
Three malformed Rhipicephalus sanguineus specimens were noticed in a tick laboratory colony. These specimens had different degrees of twining, from an almost fused individual (with two anal pores) to a partially fused adult male, with four spiracular plates. One female was a heart-shaped specimen. with both body halves fused along a longitudinal axes, with four spiracular plates and two anal pores.
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Transmission of Ehrlichia canis to dogs by ticks (Rhipicephalus sanguineus).
Two strains of Rhipicephalus sanguineus acquired Ehrlichia canis by feeding as either larvae or nymphs on acutely infected dogs and, in subsequent instars, transmitted the agent to normal dogs. Three strains of R sanguineus transmitted E canis as adults after their larval and nymphal stages fed on infected dogs. More than 400 adult female ticks were fed on infected dogs as larvae or nymphs or both, but none transmitted E canis transovarially.
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A laboratory comparison of the life cycles of the dog ticks Haemaphysalis leachi and Rhipicephalus sanguineus.
Engorged female Haemaphysalis leachi and Rhipicephalus sanguineus as well as their eggs, flat and engorged larvae and nymphs were incubated at combinations of five temperatures and three relative humidities. Mean pre-oviposition periods for H. leachi varied between 4.3 days and 12.1 days and for R. sanguineus between 4.8 days and 21.0 days. Haemaphysalis leachi converted up to 65.7% of their body mass into eggs and produced up to 16.3 eggs/mg body mass, with 4801 eggs the maximum number deposited by a single female. Peak egg production of 520 eggs/day was recorded 3 days after the commencement of oviposition. Rhipicephalus sanguineus converted up to 68.1% of their body mass into eggs and produced up to 17.2 eggs/mg body mass, with a maximum of 3,232 eggs, and peak egg production of 400 eggs/day 4 days after the commencement of oviposition. Mean incubation periods for eggs of H. leachi varied between 15.5 days and 66.7 days, and for R. sanguineus between 19.0 days and 72.0 days. Mean pre-moult periods for engorged larvae and nymphs of H. leachi varied between 14.0 days and 192.0 days, and 13.0 days and 41.0 days respectively, and for R. sanguineus between 9.5 days and 36.5 days, and 15.0 days and 44.5 days respectively. Allowing 7 days for female engorgement and 7 days for hardening of the exoskeletons and mouthparts of each of the three parasitic stages before they attach to a host, the life cycle of H. leachi would require 97-190 days and that of R. sanguineus 99-236 days to complete at the various regimes of temperature and relative humidity. The greatest proportion of H. leachi larvae engorging on mice detached between 18:00 and 19:00 on the first day of detachment and between 06:00 and 08:00 in the morning and 14:00 and 18:00 in the afternoon of the following day, while the greatest proportion of nymphs engorging on dogs detached around 19:00 on the first day of detachment and between 13:00 and 17:00 on the following day. The greatest proportion of R. sanguineus larvae engorging on dogs detached between 11:00 and 21:00 on the second day of detachment, and the greatest proportion of nymphs detached between 03:00 and 5:00 on the second and third days of detachment.
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Phenotypic and genotypic characterization of spotted fever group Rickettsiae isolated from Catalan Rhipicephalus sanguineus ticks.
Eighty-nine Rhipicephalus sanguineus ticks and 21 Rhipicephalus bursa ticks collected in Catalonia were tested by the hemolymph test to establish their infection rate with spotted fever group rickettsiae. By Giménez staining, 11.2% of the R. sanguineus isolates and 0% of the R. bursa isolates were found to contain rickettsia-like organisms. Six spotted fever group rickettsial strains (Bar29, Bar31, Gir4, Tar1, Tar2, and Tar3) were isolated from these ticks and were characterized by phenotypic and genotypic analyses. PCR followed by restriction fragment length polymorphism analysis showed that the six strains were identical and were characterized by the same restriction profiles as a strain, Mtu5, previously isolated from Rhipicephalus turanicus ticks in the South of France. Microimmunofluorescence serotyping, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified organisms, and Western blot (immunoblot) assay with mouse polyclonal sera confirmed this observation. Pulsed field gel electrophoresis of the whole genome of three of the strains showed that, although closely related, the profile of Tar1 was slightly different from that of the Bar strains. Phylogenetic analysis showed that this new rickettsial sero- and genotype, which will be named the "Catalan strain," is closely related to Rickettsia massiliae. This strain shows an unexpected resistance to rifampin. The epidemiological implications of these findings are considered.
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[In vitro pathogenicity of fungic formulation on nymphs and adults of Rhipicephalus sanguineus (Latreile, 1806) (Acari:Ixodidae)].
Rhipicephalus sanguineus is responsible for transmitting several pathogenic agents, such as: Babesia sp. and Ehrlichia sp.. This species is known as the brown dog tick and has wide geographical distribution. The purpose of this work was to evaluate the pathogenicity of biological formulations of Beauveria bassiana and Metarhizium anisopliae applied to the fed nymphs and adults of R. sanguineus under laboratory conditions. The following treatments were evaluated: control, distilled water with tween 80, emulsible concentrated, cellulose polymerized gel, fungus mixed with emulsible concentrated, fungus mixed with cellulose polymerized gel and fungus mixed with emulsible concentrated and cellulose polymerized gel. Each treatment was repeated ten times. The survival of fed nymphs and unfed adults was evaluated on the 5th, 10th, 15th and the 20th days after treatments. Significant differences were observed between the treatments (p<0.05). The treatment with M. anisopliae mixed with emulsible concentrated and cellulose polymerized gel showed the lower survival on the 15th and 20th days after treatment. We can conclude that the fungi formulation is harmful to fed nymphs and unfed adults of R. sanguineus in vitro, on this account it is suggested its use for the microbial control of this tick.
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Experimental infection models of ticks of the Rhipicephalus sanguineus group with Rickettsia conorii.
Rhipicephalus sanguineus group ticks collected on dogs in Thailand were maintained in the laboratory over several generations to test methods to infect these ticks with Rickettsia conorii, the agent of Mediterranean spotted fever. Three methods were tested: (1) infection of adults and nymphs through artificially induced bacteremic rabbit, (2) capillary feeding of solution containing 5 x 10(3) and 5 x 10(5)pfu/mL of R. conorii to adult female ticks, and (3) immersion of engorged nymphs which were "one leg-cut," "two leg-cut," or "cuticle cut" in solution containing R. conorii. The most efficient method to infect adult ticks with R. conorii was infection of ticks through the bacteremic rabbit (71.4%). The best method to infect nymphs with R. conorii was immersion of "one leg-cut" engorged nymphs in solution containing R. conorii (30%). Interestingly, a high mortality of the ticks infected with R. conorii was observed regardless of the method used. The harmful effect of R. conorii on Rh. sanguineus group ticks from Thailand is discussed including the role of the geographic origin of the ticks and the difficulties to identify ticks within this group to the species level.
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Isolation and identification of Rickettsia massiliae from Rhipicephalus sanguineus ticks collected in Arizona.
Twenty Rhipicephalus sanguineus ticks collected in eastern Arizona were tested by PCR assay to establish their infection rate with spotted fever group rickettsiae. With a nested PCR assay which detects a fragment of the Rickettsia genus-specific 17-kDa antigen gene (htrA), five ticks (25%) were found to contain rickettsial DNA. One rickettsial isolate was obtained from these ticks by inoculating a suspension of a triturated tick into monolayers of Vero E6 monkey kidney cells and XTC-2 clawed toad cells, and its cell culture and genotypic characteristics were determined. Fragments of the 16S rRNA, GltA, rOmpA, rOmpB, and Sca4 genes had 100%, 100%, 99%, 99%, and 99%, respectively, nucleotide similarity to Rickettsia massiliae strain Bar29, previously isolated from R. sanguineus in Catalonia, Spain (L. Beati et al., J. Clin. Microbiol. 34:2688-2694, 1996). The new isolate, AZT80, does not elicit cytotoxic effects in Vero cells and causes a persistent infection in XTC-2 cells. The AZT80 strain is susceptible to doxycycline but resistant to rifampin and erythromycin. Whether R. massiliae AZT80 is pathogenic or infectious for dogs and humans or can cause seroconversion to spotted fever group antigens in the United States is unknown.
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Evaluation of Rhipicephalus sanguineus as a potential biologic vector of Ehrlichia platys.
The brown dog tick, Rhipicephalus sanguineus (Acari: Ixodidae), transmits several diseases among dogs including Ehrlichia canis infection. The role of Rhipicephalus sanguineus as a biologic vector for E platys, the rickettsial agent of infectious canine cyclic thrombocytopenia, was studied in dogs. Laboratory-cultured, pathogen-free nymph ticks were fed to repletion on dogs acutely infected with E platys. Tick engorgement coincided with the development of initial parasitemia and thrombocytopenia in the infected dogs. Following repletion, nymph ticks were allowed to molt under controlled conditions. One-month-old E platys-exposed adult ticks failed to infect naive dogs in animal transmission studies. The presence of E platys was not detected in midguts or salivary glands of similarly exposed adult ticks by use of light and transmission electron microscopy. These studies indicate that R sanguineus may not transmit E platys infection.
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Rhipicephalus sanguineus (Acari: Ixodidae), the brown dog tick, parasitizing humans in Brazil.
The objective of this paper is to describe four cases of human parasitism by Rhipicephalus sanguineus (Latrielle) in Brazil. During an investigation regarding the species of ectoparasites of domestic dogs from the metropolitan region of Recife, Pernambuco state, four dog owners were found to be parasitized by ticks. The ticks were collected from these individuals and their dogs. All the ticks were identified as Rhipicephalus sanguineus . These are, to our knowledge, the first four cases of human parasitism by this tick species in Brazil. The possible implications of this finding are discussed here.
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Cutaneous hypersensitivity induced in dogs and guinea-pigs by extracts of the tick Rhipicephalus sanguineus (Acari: Ixodidae).
The cutaneous hypersensitivity induced by Rhipicephalus sanguineus tick extract in dogs (natural host) and guinea-pigs (laboratory host) was evaluated. The left ear of infested and control (tick-bite naive) dogs and guinea-pigs was injected intradermally with an extract from unfed adult ticks and the right ear with phosphate buffered saline (PBS). Ear thickness variations were then measured after 10 min and 1, 2, 6, 18, 24, 48, 72 and 96 h post-injection. Results were expressed as percentual changes in the ear thickness in relation to pre-inoculation values. The final variation in ear thickness induced by the extract was given by subtracting, in each animal, the right ear percentual increase from that of the left ear. Guinea-pigs were tested at two different times following infestation and with two different doses of extract. Infested guinea-pigs from the three experiments developed an immediate (within the first 2 h post-inoculation) and a strong delayed reaction (24 h) to the extract. Dogs, unlike guinea-pigs, developed only a strong immediate reaction whereby an 80% increase in ear thickness was observed. Control animals, with the exception of one dog, did not develop any significant reaction to the extract. Only mild reactions were induced by PBS in the right ear of all animals. The correlation between the absence of a strong delayed type reaction to tick extract and the lack of resistance of the natural host to R. sanguineus tick is discussed.
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Attempted transmission of Ehrlichia risticii, causative agent of Potomac horse fever, by the ticks, Dermacentor variabilis, Rhipicephalus sanguineus, Ixodes scapularis and Amblyomma americanum.
Dermacentor variabilis, Rhipicephalus sanguineus, Amblyomma americanum, and Ixodes scapularis ticks were investigated for their ability to transmit Potomac horse fever. Larval and nymphal ticks were exposed to Ehrlichia risticii by feeding on mice inoculated with the organism. Molted exposed ticks were then allowed to feed on susceptible ponies or mice. No evidence of transmission, either clinically or by detection of antibodies to E. risticii in mice or ponies, was observed for any tick species examined.
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Detection of Ehrlichia platys DNA in brown dog ticks (Rhipicephalus sanguineus) in Okinawa Island, Japan.
Thirty-two Rhipicephalus sanguineus females collected from eight free-roaming dogs in Okinawa Island, Japan, were examined for ehrlichial DNA by 16S rRNA-based PCR and subsequent sequencing. Partial sequences of Ehrlichia platys 16S rRNA (678 to 679 bp) were detected in three ticks (9.4%) from two dogs. This is the first report of detection of E. platys in Japan, and also the first report of detection in ticks.
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Morphological keys for the separation of the Rhipicephalus sanguineus group of ticks (Acarina:Ixodidae) in Egypt.
Rhipicephalus sanguineus sensu stricto is the taxonomic baseline for the R. sanguineus group of ticks which consists of four species in Egypt. Because of the potential confusion in the differentiation of species within the group, particularly in the Middle East, there is an urgent need for the development of new diagnostic keys. Keys, based on morphological characters, are provided herein for the various stages of R. sanguineus ticks in Egypt. An additional plan provides a useful tool for distinguishing most field collected specimens.
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Developmental profiles in tick water balance with a focus on the new Rocky Mountain spotted fever vector, Rhipicephalus sanguineus.
Recent reports indicate that the common brown dog tick, or kennel tick, Rhipicephalus sanguineus (Latreille) (Acari: Ixodidae) is a competent vector of Rocky Mountain spotted fever in the U.S.A. This tick is of concern to public health because of its high frequency of contact, as it has a unique ability to thrive within human homes. To assess the moisture requirements necessary for survival, water balance characteristics were determined for each developmental stage, from egg to adult. This is the first time that water relations in ticks have been assessed throughout the complete lifecycle. Notably, R. sanguineus is differentially adapted for life in a dry environment, as characterized by a suppressed water loss rate distinctive for each stage that distinguishes it from other ticks. Analysis of its dehydration tolerance limit and percentage body water content provides no evidence to suggest that the various stages of this tick can function more effectively containing less water, indicating that this species is modified for water conservation, not desiccation hardiness. All stages, eggs excepted, absorb water vapour from the air and can drink free water to replenish water stores. Developmentally, a shift in water balance strategies occurs in the transition from the larva, where the emphasis is on water gain (water vapour absorption from drier air), to the adult, where the emphasis is on water retention (low water loss rate). These results on the xerophilic-nature of R. sanguineus identify overhydration as the primary water stress, indicating that this tick is less dependent upon a moisture-rich habitat for survival, which matches its preference for a dry environment. We suggest that the controlled, host-confined conditions of homes and kennels have played a key role in promoting the ubiquitous distribution of R. sanguineus by creating isolated arid environments that enable this tick to establish within regions that are unfavourable for maintaining water balance.
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Experimental transmission of Dipetalonema dracunculoides (Cobbold 1870) by Rhipicephalus sanguineus (Latreille 1806).
A dog naturally infected with Dipetalonema dracunculoides and having a microfilaremia of 6050 microfilariae per mm3 of blood was used as source of infection. Experimentally cultivated nymphs of Rhipicephalus sanguineus were fed on the donor dog. Once engorged, ninety-three nymphs were removed and kept at 30 degrees C and RH 90% until they moulted to the adult stage. To study the development of microfilariae in the vector, ten ticks were dissected at Day 37 post-infection. Adult infected ticks were fed on two uninfected dogs. The observed pre-patent periods were 69 and 76 days. Trans-stadial transmission of Dipetalonema dracunculoides by Rhipicephalus sanguineus was demonstrated.
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Fine structure of the synganglion of unfed female Rhipicephalus (Rhipicephalus) sanguineus (Latreille) (Ixodoidea: Ixodidae).
The synganglion of unfed female Rhipicephalus (Rhipicephalus) sanguineus (Latreille) is enclosed within a well defined neural lamella underlined by the perineurium. No organelles were observed in the neural lamella, whereas axons containing neurosecretory vesicles, tracheal elements and glycogen deposits are visible in the perineurium. Below the perineurium, a cortical layer of nervous and non-nervous elements surrounds an inner neuropile, the latter is enclosed by a thin subperineurial layer. The neuropile is formed of nerve fibers containing neurotubules.
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