Tick Rhipicephalus Sanguineus Research Articles

An assessment of genetic variability in the mitochondrial cytochrome c oxidase subunit 1 gene of Cercopithifilaria sp. (Spirurida, Onchocercidae) from dog and Rhipicephalus sanguineus populations

This study investigates sequence variation in mitochondrial cytochrome c oxidase subunit 1 gene within Cercopithifilaria sp. recorded recently in Italy. Fourteen sequence types (haplotypes) were characterized for 163 (7.7%) amplicons from 2111 Genomic DNA samples prepared from skin samples from dogs and from Rhipicephalus sanguineus (ticks) from different geographical areas of the Mediterranean basin (i.e., Italy, Spain and Greece). The most prevalent sequence types represented haplotypes I (70.5%) and X (16.0%), followed by haplotype VIII (4.9%) and other 11 haplotypes (8.6%). Three haplotypes (II, V and VI) were found exclusively in ticks. The overall intraspecific nucleotide variation among pcox1 haplotypes ranged from 0.4 to 3.5% (mean = 1.6%), whereas a mean interspecific difference of 9.5% was detected as compared with other onchocercids. Phylogenetic analysis of the nucleotide sequence data showed a clustering of Cercopithifilaria sp. with the other Cercopithifilaria species (with strong statistical support) to the exclusion of other onchocercids. The number of haplotypes identified here might be explained by complex ecology and transmission patterns as well as the high mutation rate of mitochondrial DNA and/or inbreeding associated with hosts and their vectors.

Molecular detection of rickettsial agents in ticks and fleas collected from a European hedgehog (Erinaceus europaeus) in Marseilles, France

The European hedgehog (Erinaceus europaeus) is a synanthropic nocturnal insectivore commonly found in the countryside and in the parks and gardens. Because hedgehogs are already involved in the transmission of a number of zoonoses, including salmonellosis and ringworm, we decided to study their possible role in the epidemiology of the spotted fever group of Rickettsia. We collected ticks and fleas from a hedgehog that was captured in the city of Marseilles in France. Using a genus-specific quantitative PCR (qPCR) analysis followed by a species-specific qPCR analysis for positive samples, we observed that 91.7% (11/12) of the Rhipicephalus sanguineus ticks were positive for Rickettsia massiliae and 99.2% (128/129) of the Archaeopsylla erinacei fleas were positive for Rickettsia felis. Hedgehogs carry infected ectoparasites and then likely ensure the dissemination of spotted fever group Rickettsiae, and their epidemiological role requires further investigation.

Seroprevalence and risk factors to Ehrlichia spp. and Rickettsia spp. in dogs from the Pantanal Region of Mato Grosso State, Brazil

Sera of 320 dogs from urban and rural areas of a Pantanal region of Brazil were evaluated for rickettsial ( Rickettsia rickettsii, R. parkeri, R. amblyommii, R. rhipicephali, R. felis, and R. bellii) and ehrlichial ( Ehrlichia canis) infection by the immunofluorescence assay (IFA). Risk factors for ehrlichiosis or rickettsiosis were also evaluated. Positive reaction against Ehrlichia spp. was detected in 227 (70.9%) dogs, 119 (74.3%) from an urban area and 108 (67.5%) from rural areas ( P > 0.05). For Rickettsia spp., 152 (47.5%) dogs were positive, 31 (19.3%) from urban and 121 (75.6%) from rural areas ( P < 0.05). Highest anti- Rickettsia spp. endpoint titers were observed for R. amblyommii, suggesting homologous reactions to this agent or a very closely related organism. While most of the urban dogs were found parasitized by the tick Rhipicephalus sanguineus, infestations on rural dogs were predominated by Amblyomma cajenennse. Rickettsial infection was significantly higher ( P < 0.05) in rural dogs, in dogs with hunting practice, or in A. cajennense-infested dogs.

Domestic Dogs (Canis familiaris) as Reservoir Hosts forRickettsia conorii

Rickettsia conorii is the causative agent of Mediterranean spotted fever (MSF) and Israeli spotted fever (ISF) transmitted by the brown dog tick Rhipicephalus sanguineus. In areas where MSF or ISF are prevalent, dogs have high prevalence of R. conorii -neutralizing antibodies. However, the true role of dogs in the persistence of the R. conorii transmission cycle is unknown, and their reservoir competence for this pathogen has remained untested. We assessed the ability of dogs infected with R. conorii to transmit the pathogen to previously uninfected Rh. sanguineus ticks. Dogs were infected either via needle-inoculation of cultured rickettsiae or naturally via infected tick bite. Dogs were monitored for clinical signs of infection, for rickettsemia by PCR, and for seroconversion and were subjected to infestation with uninfected ticks at different time points. Rh. sanguineus larvae and nymphs successfully acquired the agent from both needle-inoculated and tick-infected dogs and transmitted it transtadially. Tick-infected dogs remained infectious to ticks for at least a month postinfection. The molted ticks were, in turn, infectious to naïve dogs. These results demonstrate that dogs are capable of acquiring R. conorii from infected Rh. sanguineus ticks and transmitting infection to cohorts of uninfected ticks, thus confirming for the first time that dogs are indeed competent reservoirs for R. conorii. In addition, dogs with different genetic backgrounds appear to differ in their susceptibility to R. conorii infection.

Effects of aggregation on the reproductive biology of Rhipicephalus sanguineus females

The brown dog tick Rhipicephalus sanguineus is one of the most important tick species in both veterinary and human medicine. Studies on the biology of this tick are pivotal in order to develop improved control strategies. The brown dog tick is a gregarious species, tending to form large clusters in highly infested environments. This is the case of engorged females of R. sanguineus that often tend to aggregate in the environment. In the present study, we assessed whether aggregation affects R. sanguineus females in the laboratory. Engorged females of R. sanguineus were separated in six groups, each with four replicates. Six groups (G1-G6) were composed of single, five, 10, 20, 40 or 60 females, for a total of 544 females. We found that aggregation (expressed as tick density within Petri dishes) affected egg production efficiency of females and incubation period, but not egg hatching. In particular, there was a strong negative correlation between tick density and the following parameters: egg mass weight, incubation period, egg production efficiency, reproduction efficiency index, and reproductive fitness index. However, for the maintenance of tick colonies, engorged females can be maintained in groups (density up to 0.4/cm(2)) without major prejudices in terms of egg production and hatch rate. This information might be useful for those interested in maintaining large tick colonies, as grouping engorged females might help to save space in incubators and, most importantly, to keep the genetic heterogeneity within the tick colony, across generations.

Mediterranean spotted fever and encephalitis: a case report and review of the literature

Mediterranean spotted fever (MSF) is a disease caused by Rickettsia conorii and transmitted by the brown dog tick Rhipicephalus sanguineus. It is widely distributed through southern Europe, Africa, and the Middle East. It is an emerging or a reemerging disease in some regions. Countries of the Mediterranean basin, such as Portugal, have noticed an increased incidence of MSF over the past 10 years. It was believed that MSF was a benign disease associated with a mortality rate of 1-3% before the antimicrobial drug era. It was called benign summer typhus. Severe forms were described in 1981, and the mortality rate reached 32% in Portugal in 1997. However, neurological manifestations associated with brain lesions are a rare event. We describe the case of a man with fever, maculopapular rash, a black spot, and hemisensory loss including the face on the left side of the body with brain lesions in the imaging studies.

Serodetection of Ehrlichia canis infection in dogs from Ludhiana district of Punjab, India

Dot-ELISA (enzyme-linked immunosorbent assay) Immunocomb(®) assay was conducted to detect the presence of antibodies against Ehrlichia canis in blood samples of 60 privately owned dogs suspected to be infected with E. canis from the Small Animal Clinics, College of Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab (India). Antibodies reactive to E. canis were detected in plasma in 48 samples out of 60 samples by Immunocomb(®) Dot-ELISA. Out of these 39.58% samples were low positive (Titre 1:20-1:40), 31.25% were medium positive (Titre 1:80-1:640) and 29.16% were high positive (Titre>1,280), for the infection. When examined by microscopy, only two samples revealed typical E. canis morulae. Haemato-cellular examination revealed thrombocytopenia along with anaemia and leucopenia. Results suggest that E. canis infection circulates in dogs in India in low non-detectable numbers by microscopy and is transmitted by the brown dog tick Rhipicephalus sanguineus.

Rhipicephalus sanguineus (Ixodida, Ixodidae) as intermediate host of a canine neglected filarial species with dermal microfilariae

The life cycles of filarioids of dogs presenting dermal microfilariae have been little studied. Following the recent retrieval of dermal microfilariae identified as Cercopithifilaria sp. in a dog from Sicily (Italy), this study was designed to assess the role of the brown dog tick Rhipicephalus sanguineus as an intermediate host of this filarial species. An experimental tick infestation was performed on an infected dog using 300 nymphs of R. sanguineus. Engorged nymphs were collected and examined by both microscopic dissection and molecular analysis at five time points (i.e., the same day of tick detachment and 10, 20, 30 and 50 days post-detachment) to detect the presence and developmental stage of filariae in the ticks. A total of 270 engorged nymphs were collected from the dog and developing filarioid larvae detected in 10 (5%) out of 200 ticks dissected. Infective third-stage larvae were observed in 4 (2%) of the all dissected ticks, 30 days post-detachment. Twelve (6.6%) out of 181 samples molecularly tested were positive for Cercopithifilaria sp. This study demonstrates that nymphs of R. sanguineus feeding on a dog naturally infected by Cercopithifilaria sp. can ingest microfilariae, which develop up to the third infective stage thus suggesting that this tick species might act as an intermediate host of this little known canine filarioid.

Efficacy of a novel topical combination of fipronil, amitraz and (S)-methoprene for treatment and control of induced infestations of brown dog ticks ( Rhipicephalus sanguineus) on dogs

Four laboratory studies were conducted to demonstrate that a single topical dose of a novel spot-on combination containing fipronil, amitraz and (S)-methoprene (CERTIFECT™, Merial Limited, GA, USA) is efficacious against the brown dog tick, Rhipicephalus sanguineus. In each study, 6–8 male and 6–8 female purpose-bred, laboratory mongrels, terrier cross or Beagles were randomly assigned to one of two study groups (treated and untreated), based on pre-treatment parasite counts. Starting on the day before treatment, each dog was infested weekly with 50 ticks. Ticks were thumb counted at various time points after treatment and weekly infestations starting as early as 6 h and continued at 12, 18 and 24 h depending on the study. Ticks were removed and counted at 48 h after treatment and weekly infestations. CERTIFECT provided rapid and excellent control of pre-existing and newly acquired infestations of R. sanguineus with efficacy as high as 93% within the first 12 h after a single topical treatment. Excellent control (>96%) of R. sanguineus as early as 18 h, following post treatment infestations was maintained for at least 35 days.

Israeli Spotted Fever, Tunisia

Israeli Spotted Fever, Tunisia

Rickettsia,Ehrlichia,Anaplasma, andBartonellain Ticks and Fleas from Dogs and Cats in Bangkok

Flea and tick specimens (5-10 fleas or ticks) on dogs and cats from various sites in Bangkok were tested by polymerase chain reaction and DNA sequencing to detect DNA of bacteria Rickettsia (gltA and 17 kDa genes), Anaplasmataceae (16S rRNA gene), and Bartonella (pap31 and its genes). We confirmed that Rickettsia sp. related to Rickettsia felis was detected in 66 of 98 (67.4%) flea specimens from dogs, whereas 8 Bartonella henselae and 2 Bartonella clarridgeiae were detected in 10 of 54 (18.5%) flea specimens from cats. Further, this work provides the first evidence of 10 Ehrlichia canis (3.3%), 7 Anaplasma platys (2.3%), and 2 Wolbachia spp. (0.66%) in 304 Rhipicephalus sanguineus tick specimens in Thailand.

Tick Infestation and Spotted‐Fever Group Rickettsia in Shelter Dogs, California, 2009

In response to an outbreak of Rocky Mountain spotted fever (RMSF) in Baja California in early 2009, dogs at two shelters in neighbouring Imperial County, California, were evaluated for ectoparasites. Brown dog ticks (Rhipicephalus sanguineus), a recognized vector for RMSF, were found on 35 (30%) of 116 dogs but all ticks tested negative for Rickettsia rickettsii by PCR.

Mathematical modelling of the impact of climatic conditions in France on Rhipicephalus sanguineus tick activity and density since 1960

Rhipicephalus sanguineus, the brown dog tick, has a worldwide distribution in areas with a relatively warm climate, including mild winters. This tick plays an important role as vector for various animal and human pathogens, including bacteria and protozoa. Based on precise daily meteorological data from the past 40 years, combined with mathematical modelling designed to predict tick activity, two modelling approaches were developed. The first examined the evolution of the number of weeks with favourable biological conditions for ticks in four French cities located at various latitudes of the country: Nîmes in the south, Paris in the north, Lyon in the east and Nantes in the west. The second analysed the extension of the geographical surface area in km(2) where the biological conditions favour tick activity for at least 12 weeks per year. Both analyses revealed clear evidence of increased temperatures coupled with an augmented tick activity index in three of the four cities. However, the change was not significant in Nîmes, where the climate is Mediterranean and the tick is already endemic. For Paris, Lyon and Nantes, the activity index values have increased significantly, i.e. by 4.4%, 4.0% and 3.4%, respectively. The distribution of the activity index values is evolving strongly with significantly fewer values below 50% since the 1960s and a clear decrease of values between 20% and 50% during the latest decade. Between 1960 and 2000, the theoretical extension of the surface area where the climatic index is suitable for R. sanguineus has increased by 66%. Even though several other important factors, such as changes in biotopes or human activity, are not included in this study, the resulting patterns and trends are noticeable. Our models constitute the first demonstration of the impact of climate change on the activity and distribution of ticks and confirm the observed northward migration trend for this Mediterranean domestic tick.

Deconstructing Tick Saliva: NON-PROTEIN MOLECULES WITH POTENT IMMUNOMODULATORY PROPERTIES

Dendritic cells (DCs) are powerful initiators of innate and adaptive immune responses. Ticks are blood-sucking ectoparasite arthropods that suppress host immunity by secreting immunomodulatory molecules in their saliva. Here, compounds present in Rhipicephalus sanguineus tick saliva with immunomodulatory effects on DC differentiation, cytokine production, and costimulatory molecule expression were identified. R. sanguineus tick saliva inhibited IL-12p40 and TNF-α while potentiating IL-10 cytokine production by bone marrow-derived DCs stimulated by Toll-like receptor-2, -4, and -9 agonists. To identify the molecules responsible for these effects, we fractionated the saliva through microcon filtration and reversed-phase HPLC and tested each fraction for DC maturation. Fractions with proven effects were analyzed by micro-HPLC tandem mass spectrometry or competition ELISA. Thus, we identified for the first time in tick saliva the purine nucleoside adenosine (concentration of ∼110 pmol/μl) as a potent anti-inflammatory salivary inhibitor of DC cytokine production. We also found prostaglandin E(2) (PGE(2) ∼100 nM) with comparable effects in modulating cytokine production by DCs. Both Ado and PGE(2) inhibited cytokine production by inducing cAMP-PKA signaling in DCs. Additionally, both Ado and PGE(2) were able to inhibit expression of CD40 in mature DCs. Finally, flow cytometry analysis revealed that PGE(2), but not Ado, is the differentiation inhibitor of bone marrow-derived DCs. The presence of non-protein molecules adenosine and PGE(2) in tick saliva indicates an important evolutionary mechanism used by ticks to subvert host immune cells and allow them to successfully complete their blood meal and life cycle.

Synergy between fipronil and amitraz in a Rhipicephalus sanguineus tick residual contact test

Amitraz, a formamidine acaricide, has been reported to act as a synergist of contact insecticides and acaricides in both agriculture and animal health. A laboratory contact test was therefore conducted to determine whether amitraz at 12.5 ppm could improve the contact potency of fipronil on ticks. A controlled glass vial bioassay was used to assess the efficacy of fipronil alone, amitraz alone, and fipronil plus amitraz on unfed adult Rhipicephalus sanguineus. Assessments of lethality were made at 6, 24, and 48 h after the introduction of the ticks to the vials. No significant mortality was observed in the control treatment or in the amitraz alone treatment. Concentration and time dependent mortality rates were observed in ticks exposed to fipronil alone or fipronil plus amitraz, with higher mortality observed in the latter group. Results from this study gave synergistic EC50 ratios between fipronil alone and fipronil plus amitraz of >7.3, 137 and 97 at 6, 24, and 48 h, respectively. A similar response was seen at the EC90 level. These results indicate that fipronil was synergized by amitraz in this adult tick residual contact study. The addition of amitraz to fipronil also provided a significant improvement in the speed of kill.

Infection by Spotted Fever Rickettsiae in People, Dogs, Horses and Ticks in Londrina, Parana State, Brazil

Spotted fever is a disease caused by bacteria from the genus Rickettsia of the spotted fever group (SFG). Rickettsia rickettsii is likely the main agent of Brazilian spotted fever (BSF). With the objective of gathering information on the circulation of SFG rickettsiae in Londrina, Parana state, ticks from dogs and horses and also blood from dogs, horses and humans were collected in a neighbourhood of the city which presented potential for circulation of rickettsiae between hosts and vectors. Amblyomma cajennense, Dermacentor nitens, and Rhipicephalus sanguineus ticks were subjected to Polymerase Chain Reaction targeting a fragment of the Rickettsia gltA gene. This specific gene encodes the enzyme citrate synthase of Rickettsia spp., and results on all ticks were negative. Human and animal sera were tested by Indirect Immunofluorescence Assay in which R. rickettsii and R. parkeri were used as antigens. Sera from 4.7% human, 2.7% canine and 38.5% equine were positive for R. rickettsii. For R. parkeri, 0.9% human, 2.7% canine and 11.5% equine samples were positive. All samples reactive to R. parkeri also reacted to R. rickettsii. An epidemiological questionnaire was applied, but there were no statistically significant results. Comparison of our serological results with previous studies in Brazil, among BSF endemic and non-endemic areas, indicates that there is no established rickettsial infection in the study area, a statement corroborated with our molecular analysis. Nonetheless, as humans of the present study are highly exposed to tick infestations, health education within the population is needed to obtain efficient tick control.

Inhibitory action of neem aqueous extract (azadirachta indica A. Juss) on the vitellogenesis of rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) ticks

The present study revealed unheard of data about the action of aqueous extracts of neem leaves (Azadirachta indica) on the vitellogenesis of Rhipicephalus sanguineus ticks, proving that these extracts in 10 and 20% concentrations do not have the potential to kill the females; however, in lower concentrations (10%) provokes great morphological alterations in germinative cells such as the emergence of extended cytoplasmic vacuolization areas as well as the fragmentation of the germinal vesicle, even in those oocytes which were in initial stages of development (I-III), showing that neem is a potent agent which acts impeding one of the main metabolic stages of the ticks, i.e., the reproduction. In oocytes in final stages of development (IV-V) azadirachtin (neem's active principle) caused significant reduction in the size and quantity of proteic granules of the yolk and the inversion of their localization where the smaller granules before inside the cell (normal oocyte) were posteriorly observed in the periphery, and the bigger ones in the central region. Thus, the study showed that the alterations found both in the oocytes and in the pedicel cells indicated that azadirachtin acts on the process of tick's reproduction and signalizes that this plant can be used in the future to control ticks with the advantage of not being aggressive to nontarget organism or the environment. Furthermore, data here obtained showed that the most significant efficiency of the aqueous extract of neem is related to the concentration of 10%, proving that higher doses would not be so efficient.

Rickettsial infection in domestic mammals and their ectoparasites in El Valle de Antón, Coclé, Panamá

The present research evaluated the presence of Rickettsia spp. on ectoparasites of horses and dogs (using PCR techniques), and their sera (using immunofluorescence assay) in El Valle de Antón town in Panama. A total of 20 horses and 20 dogs were sampled, finding four species of ectoparasites on dogs (the ticks Rhipicephalus sanguineus, Amblyomma ovale, Amblyomma oblongoguttatum, and the flea Ctenocephalides felis), and two tick species on horses (Amblyomma cajennense and Dermacentor nitens). DNA of Rickettsia amblyommii was found in pools of A. cajennense, D. nitens, and R. sanguineus, while Rickettsia felis was detected in C. felis pools. Overall, 70% (14/20) and 65% (13/20) of the horses and dogs, respectively, were seroreactive (titer ≥64) to spotted fever group rickettsiae. Sera from six dogs and five horses reacted to R. amblyommii antigens with titers at least four-fold higher than those for the other antigens tested (Rickettsia bellii, Rickettsia parkeri, Rickettsia rhipicephali, R. felis, and R. rickettsii). These serological results, coupled with our molecular findings, suggest that these dogs and horses were infected by Rickettsia amblyommii. More studies need to be realized afford to identify the Rickettsia species responsible for other serological and molecular positive results, and their ecological importance.

Effects of Ricinus communis oil esters on salivary glands of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae)

This study showed the interference of esters extracted from Ricinus communis in the secretory cycle of salivary glands of Rhipicephalus sanguineus ticks, which consequently caused collateral effects on their feeding process. Ticks attached on hosts which were fed with commercial feed containing different concentrations of R. communis oil esters suffered damages such as cytoplasmic changes in their salivary glands, notably in the acinar cells, impairing the functioning of the acini and accelerating the organs degeneration as a whole. It was found that esters interfered with the activity of cellular secretion by changing the glycoprotein of salivar composition especially in acini II cells. It was also shown that the damages caused by esters in the salivary glands cells of these ectoparasites increased in higher concentrations of the product and degenerative glandular changes were more pronounced.

Ectoparasitos de animais silvestres no Maranhão

Objetivou-se identificar os ectoparasitos de animais silvestres recepcionados pelo Centro de Triagem de Animais Silvestres, São Luís, Maranhão. Os ectopararasitos identificados foram: piolhos Acidoproctus sp., Trinoton sp., Ciconiphilus sp, Austromenopon sp., Quadraceps sp., Saemundssonia sp. e Trichodectes canis; carrapato Amblyomma rotundatum e Rhipicephalus sanguineus; pulgas Ctenocephalides felis e larvas de Cochliomyia hominivorax. Os resultados apresentados documentam a infestação de mamíferos, répteis e aves silvestres por ectoparasitos.