Participation of Rhipicephalus sanguineus (Acari: Ixodidae) in the epidemiology of canine visceral leishmaniasis

The vectorial competence of the tick Rhipicephalus sanguineus is discussed in relation to the epidemiology of canine visceral leishmaniasis, taking into account its strict association with dogs and the low indices of natural infection presented by its known vector, the phlebotomine sand fly Lutzomyia longipalpis. In order to evaluate natural infection by Leishmania chagasi and the infectivity of these parasites in the tick, 39 specimens (6 females, 11 males and 22 nymphs) of R. sanguineus were removed from 21 dogs showing diverse symptoms of zoonotic visceral leishmaniasis (ZVL). Six ticks (15.4%) gave positive results for the genus Leishmania using the PCR technique. To determine the infectivity of the parasites, 36 hamsters were inoculated orally and peritoneally with macerates of ticks removed from nine dogs symptomatic for visceral leishmaniasis. After 6 months the hamsters were sacrificed and necropsied. Serum was removed for IFAT, as well as spleen and liver fragments to make imprint smears and for PCR. Eight (88.9%) of these dogs presented ticks that were infective for 14 hamsters (41.2%), 12 (85.7%) of them infected peritoneally and two (14.3%) orally. PCR revealed 27 smears (40.9%) to be positive, 20 (62.5%) of them infected peritoneally and seven (20.6%) orally. IFAT showed 14 positive animals (41.2%). Based on these findings, we suggest that the vectorial capacity of R. sanguineus for L. chagasi should be evaluated further, opening new perspectives in the epidemiology of ZVL.     

Apparent tick paralysis by Rhipicephalus sanguineus (Acari: Ixodidae) in dogs

Certain tick species including Ixodes holocyclus can inoculate neurotoxins that induce a rapid, ascending flaccid paralysis in animals. Rhipicephalus sanguineus, the most widespread tick of dogs, is recognized as a vector of several pathogens causing diseases in dogs and humans. A single report suggests its role as cause of paralysis in dogs. This study presents the clinical history of 14 young dogs heavily infested by R. sanguineus (intensity of infestation, 63-328) in an endemic area of southern Italy. During May to June of 2011, dogs were presented at the clinical examination with neurological signs of different degrees (e.g., hind limb ataxia, generalized lethargy, and difficulty in movements). All animals were treated with acaricides and by manual tick removal but ten of them died within a day, displaying neurological signs. The other 4 dogs recovered within 3 days with acaricidal and supportive treatment. Twelve dogs were positive by blood smear examination for Hepatozoon canis with a high parasitemia, two also for Babesia vogeli and two were negative for hemoparasites. Low-grade thrombocytopenia, hypoalbuminemia, and pancytopenia were the haematological alterations most frequently recorded. Other causes of neurological disease in dogs were excluded and the diagnosis of tick paralysis by R. sanguineus was confirmed (ex juvantibus) by early and complete recovery of 4 dogs following acaricidal treatment and tick removal.     

Degeneration of salivary glands of males of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari, Ixodidae)

The present study examined the salivary glands of Rhipicephalus sanguineus males at days 0, 3, and 7 post-detachment from the host. Degeneration of this organ occurred in the three stages and it advanced as time away from the host progressed. Thus, characteristics of degeneration were more prominent in males at day 7 post-detachment than in males at day 0 post-detachment. In males at day 0 post-detachment, type I acini were intact; while in other stages these acini exhibited signs of degeneration. In type II acini of individuals at day 0 post-detachment, cells a, c1-c5, c8, and indeterminate were identified. Only c1 and c8 were intact. The remaining cell types were undergoing degeneration, as well as all cells d-f in type III acini, and all g in type IV acini. In males at day 3 post-detachment from the host, all cells (a, c1-c5, c8 and indeterminate) of type II acini, cells d and e in type III acini, and g in type IV were undergoing degeneration. In some Indeterminate acini, the boundaries of cells still could be distinguished, while in others, only a cytoplasmic mass was observed. At day 3 post-detachment, apoptotic bodies were present. In males at day 7 post-detachment from the host, the degeneration process progressed. All cells a, c1, c3-c5, c8 and indeterminate in type II, and d and e in type III acini were undergoing degeneration. Type IV acini still contained remnants of secretion and in Indeterminate acini, only a cytoplasmic mass could be observed. At this stage, apoptotic bodies were also present. The present study still revealed that cells of salivary glands of R. sanguineus males when degenerating undergo the following changes: (a) decrease in secretion production with or without granule breakage, (b) changes in nuclear morphology, (c) cytoplasm shrinkage, (d) loss of cell shape, (e) loss of cell boundaries, and (e) cytoplasmic vacuolation. Together, these changes result in cell fragmentation with release of apoptotic bodies.     

First evidence of Anaplasma platys in Rhipicephalus sanguineus (Acari: Ixodida) collected from dogs in Africa

A total of 27 ticks, comprising Rhipicephalus sanguineus (Latreille) (n = 21), Haemaphysalis leachi (Andouin) (n = 4) and Haemaphysalis paraleachi (Camicas, Hoogstraal & El Kammah) (n = 2) were recovered from two clinically healthy female dogs in the Democratic Republic of the Congo. DNA of Anaplasma platys was detected in a female R. sanguineus, using primers derived from the 16S rRNA gene, which amplify members of the family Anaplasmataceae . Anaplasma platys DNA was also detected in the blood of one of the dogs. Phylogenetic analysis based on partial sequences of the 16S rRNA, the gltA and the groEL genes ranged the detected agent within the Anaplasma clade. This is the first reported detection of A. platys in ticks in Africa. This finding raises the question of the possible involvement of R. sanguineus in A. platys infection of dogs.     

The brown dog tick, Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae): from taxonomy to control

Rhipicephalus sanguineus, commonly known as the brown dog tick, is a three-host tick that feeds primarily on dogs and occasionally on other hosts, including humans. R. sanguineus ticks are widely distributed around the world and they are known vectors of pathogens, such as Babesia canis, Ehrlichia canis, and Rickettsia conorii. The increasing number of cases of human parasitism by R. sanguineus ticks reported in the literature indicates that the interaction between humans and R. sanguineus ticks may be more common than it is actually recognized. The indiscriminate use of acaricides is an emerging problem worldwide and has led to the selection of acaricide resistant tick strains. In this article, the medical and veterinary importance, taxonomy, biology, and ecology of R. sanguineus ticks around the world are reviewed. It also discusses the current strategies for the control of R. sanguineus, highlighting the potential risks associated to the improper use of acaricides, such as environmental pollution and toxicity to humans and other non-target organisms (e.g., tick predators).     

Acaricidal activity of hydroethanolic formulations of thymol against Rhipicephalus sanguineus (Acari: Ixodidae) and Dermacentor nitens (Acari: Ixodidae) larvae

The aim of this study was to assess the acaricidal activity of hydroethanolic formulations of thymol at varying concentrations on Rhipicephalus sanguineus and Dermacentor nitens larvae. The larval packet test was used and the thymol concentrations tested were 2.5, 5.0, 10.0, 15.0 and 20.0 mg/ml. The control group was exposed only to water and ethanol (50/50%) and there were 10 repetitions for each treatment. The mortality was evaluated after 24 h. For the R. sanguineus larvae, the mortality rates were 47.5, 50.2, 96.7, 95.9 and 98.1%, while for D. nitens the rates were 14.1, 75.0, 90.2, 90.3 and 99.5%, at respective thymol concentrations of 2.5, 5.0, 10.0, 15.0 and 20.0 mg/ml. The results indicate that the hydroethanolic formulations of thymol tested have acaricidal activity on R. sanguineus and D. nitens larvae exposed topically, causing mortality greater than 90% 24 h post-treatment starting at the concentration of 10 mg/ml.     

Comparison of the external morphology of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) ticks from Brazil and Argentina

In the present study the external morphology of semi-engorged Rhipicephalus sanguineus females ticks from Brazil and Argentina were compared by scanning electron microscopy. Many differences were detected among the R. sanguineus collected at the two localities, such as body size, shape of the genital pore, and morphology of the sensory structures. All these characteristics are fundamental for the diagnosis of species of the genus Rhipicephalus and thus indicate the need for further comparisons and, the taxonomical revision of this species of tick in the Neotropics.     

Efficacy of the Bm86 antigen against immature instars and adults of the dog tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae)

The Bm86 antigen has been used to control ticks of the Boophilus genera in integrated programs that also include the use of acaricides. Because of recent phylogenetic studies have lead to the inclusion of all Boophilus species within the Rhipicephalus genera, we aimed to investigate the efficacy of the Bm86 antigen on the biotic potential of Rhipicephalus sanguineus. Domestic dogs were vaccinated with Bm86 and challenged with the three instars of R. sanguineus. Male and female mongrel dogs were divided into two groups of four animals each, comprising non-vaccinated and vaccinated animals. Immunized dogs were given two doses of an experimental formulation containing 50 μg of recombinant Bm86, at 21 days interval while the other group was given placebo, consisting of the same preparation without Bm86. Each dog was challenged 21 days after the last dose with 250 larvae, 100 nymphs and 55 adults (25 females and 30 males) released inside feeding chambers (one per instar) glued to their shaved flank. The effect of the vaccination was evaluated by determining biological parameters of ticks including the yield rates of larvae, nymphs and adult females. Adult females engorged weight, egg mass weight, efficiency rate of conversion to eggs (ERCE) and hatchability. In addition, sera were collected from dogs at 0, 21, 36, 45 and 75 days after the vaccination and used for the detection of specific antibodies by ELISA. Collection rates of larvae, nymphs and adult females fed on vaccinated dogs were significantly (p < 0.05) reduced by 38%, 29% and 31%, respectively, as compared with non-vaccinated controls. Significant reductions were also observed in weight of engorged females and egg mass, in ERCE, but not in the hatch rate of ticks fed on immunized dogs. ELISA data revealed a marked and significant increase in optical densities of sera from vaccinated animals after the second dose of Bm86. We concluded that the Bm86 antigen used as a vaccine for dogs reduced the viability and biotic potential of the R. sanguineus.     

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.     

Biological and DNA evidence of two dissimilar populations of the Rhipicephalus sanguineus tick group (Acari: Ixodidae) in South America

In this work, the biology, mitochondrial DNA and fertility of hybrids from two strains of Rhipicephalus sanguineus, from Brazil and Argentina, were compared. Engorged larvae, nymphs and adults from Argentina weighed more and the engorgement period of adult females was significantly longer than those of their Brazilian counterparts, whereas adult female tick yield rate was higher for the Brazilian strain. High intraspecific divergence of mitochondrial DNA was detected between R. sanguineus from Brazil and Argentina. On the other hand, a strong genetic relationship was detected between European and Argentinean R. sanguineus populations while the Brazilian population appeared to be related to the African Rhipicephalus turanicus. Adult hybrid females laid eggs, which were mostly unviable, whereas a mean of more than 1400 larvae hatched per egg mass from pure Brazilian and Argentinean strains. These results showed that differences between these strains are greater than previously assumed and that the biosystematic status of R. sanguineus ticks from South America should be re-evaluated. Wide variations, such as these might account for the reported worldwide differences in biology and vector capacity of this species.     

Effect of ricinoleic acid esters from castor oil (Ricinus communis) on the oocyte yolk components of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae)

Rhipicephalus sanguineus are bloodsucking ectoparasites, whose main host is the domestic dog, thus being present in urban areas and closely located to people. Eventually, this tick species parasitize humans and can become a potential vector of infectious diseases. Methods to control this type of pest have been the focus of many research groups worldwide. The use of natural products is increasingly considered nowadays, due to the low toxicity levels to the host and low waste generation to the environment. This study tested the effect of ricinoleic acid esters from castor oil (as an potential acaricide) on the reproductive system of R. sanguineus females, more specifically on the vitellogenesis process. For this, two groups were established: the control group (CG) and the treatment group (TG) with five rabbits in each (New Zealand White), used as hosts. NaCl and ester were added to rabbits’ food and offered to the hosts. After full engorgement, the females were collected and had their ovaries extracted. The ticks ovaries were submitted to histochemical techniques so the effects of esters could be observed over polysaccharides, proteins and lipids yolk. Changes in the deposition of yolk components were observed. This caused modifications on elements of polysaccharide origin and on glycoprotein compounds, interfering in the final yolk synthesis and compromising the development of the future embryo.     

Acaricidal activity of Calea serrata (Asteraceae) on Boophilus microplus and Rhipicephalus sanguineus

Calea serrata (Asteraceae) is an endemic Southern Brazilian plant species used for religious and medicinal purposes. Previous study revealed the presence of chromenes, a class of natural compounds that possess insecticidal properties. This study reports the effect of the hexane extract from the aerial parts of this plant on egg hatchability, egg production and mortality rates of newly hatched larvae of the cattle tick Boophilus microplus. Larvae of Rhipicephalus sanguineus, the brown dog tick, received the same treatment. The extract was toxic to the eggs of B. microplus and to the larvae of both B. microplus and R. sanguineus.     

Discrimination between six species of canine microfilariae by a single polymerase chain reaction

Canine dirofilariasis caused by Dirofilaria immitis is usually diagnosed by specific antigen testing and/or identification of microfilariae. However, D. immitis and at least six other filariae can produce canine microfilaremias with negative heartworm antigen tests. Discriminating these can be of clinical importance. To resolve discordant diagnoses by two diagnostic laboratories in an antigen-negative, microfilaremic dog recently imported into the US from Europe we developed a simple molecular method of identifying different microfilariae, and subsequently validated our method against six different filariae known to infect dogs by amplifying ribosomal DNA spacer sequences by polymerase chain reaction using common and species-specific primers, and sequencing the products to confirm the genotype of the filariae. We identified the filaria in this dog as D. repens. This is the first case of D. repens infection in the United States. Additionally, we examined microfilariae from five additional antigen-negative, microfilaremic dogs and successfully identified the infecting parasite in each case. Our diagnoses differed from the initial morphological diagnosis in three of these cases, demonstrating the inaccuracy of morphological diagnosis. In each case, microfilariae identified morphologically as A. reconditum were identified as D. immitis by molecular methods. Finally, we demonstrated that our PCR method should amplify DNA from at least two additional filariae (Onchocerca and Mansonella), suggesting that this method may be suitable for genotyping all members of the family Onchocercidae.     

Efficacy of amitraz applied as a dip against an amitraz-resistant strain of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) infested on cattle

Selection pressure with the acaricide, amitraz was applied to a strain of Rhipicephalus (Boophilus) microplus (Canestrini) in 13 out of 18 generations, resulting in a 28.7-fold increase in resistance, indicating a shift in the phenotypic composition of the ticks, from a heterogenous mixture of both susceptible and resistant ticks when selection began to a homogenous composition of predominantly resistant ticks when selection ended. Resistant larvae of the last selected generation were infested on cattle three separate times at 20, 13, and 6 d (7 d intervals) before being dipped in a total immersion vat charged at 0.02, 0.044, and 0.096% active ingredient (AI) amitraz. There was a dose related, though not always significant, response to amitraz whereby each increase in concentration produced increased adverse effects. The number of ticks and index of fecundity (IF) of females recovered from cattle treated at 0.02 and 0.044% AI were not different from that of untreated ticks. However, female weight and egg mass weight of females recovered from cattle treated at 0.044% were lower than untreated females or females treated at 0.02% AI. All measured parameters obtained from ticks recovered from cattle treated at 0.096% AI were significantly lower than the untreated control group, but tick numbers and IF were not different from the other treated groups. At 0.02% AI the level of control was significantly lower (34.6%) than treatment at 0.096% AI (81.2%), while the level of control at 0.044% AI (57.6%) was intermediate between the other treated groups. Therefore, none of the amitraz treatments provided the required 99% control necessary for use in the U.S. Cattle Fever Tick Eradication Program, and the estimated line derived from regression analysis showed that a concentration of ca. 0.2% AI amitraz would be needed to achieve 99% control of the resistant ticks.