Ehrlichia canis TRP36 diversity in naturally infected-dogs from an urban area of Colombia.

Ehrlichia canis is the etiologic agent of a highly prevalent tick-borne disease, canine monocytic ehrlichiosis (CME). Four defined E. canis genotypes based on the trp36 gene sequences have been reported, three of them identified in North or South America. The diversity of E. canis has been investigated using genetic and serologic approaches based on distinct 36 kDa tandem repeat protein (trp36) gene sequences that have been reported. The main objectives of this study were to determine the prevalence of E. canis infection in dogs from Medellín, Colombia by PCR and determine the E. canis diversity using molecular and serologic approaches. Blood was collected from dogs (n = 300) with clinical signs of CME for PCR detection of E. canis 16S rRNA, dsb and trp36 DNA. Phylogenetic analysis of trp36 gene sequences was performed using MEGA. A serological evaluation was performed using immunofluorescence microscopy and ELISA with species-specific peptides from E. canis TRP19 and TRP36 (3 genotypes) and E. chaffeensis (TRP32). E. canis DNA (16S rRNA and/or dsb) was detected in 18 % (53/300) of dogs by PCR amplification. The trp36 gene was amplified and sequenced from 35/53 16S rRNA/dsb PCR positive samples revealing three genotypes: United States (US; n = 21), Costa Rica (CR; n = 11), and Brazil (BR; n = 3). Most dogs (33/35) with detectable trp36 DNA had anti-E. canis TRP19 and TRP36 peptide antibodies that corresponded to the genotype detected by PCR. Dogs that had antibodies to the TRP19 peptide (82/300; 38 %), also had antibodies to one or more genotype-specific TRP36 peptides. Based on TRP36 serology, the dogs exhibited highest frequency of infection with the US genogroup (US = 26), followed by the CR genogroup (CR = 19) and the BR genogroup (BR = 11). Notably, 26/53 trp36 PCR positive dogs had detectable antibodies to multiple E. canis genotypes (US/BR/CR = 8, BR/CR = 7, US/CR = 6 and US/BR = 5) suggesting coinfection or multiple sequential infections with different genotypes. Colombian dogs did not have antibodies to E. chaffeensis as determined by a TRP32 species-specific ELISA. Our results demonstrate the presence of three previously defined genotypes in North and South America in Colombian dogs (US, BR, CR). These results also demonstrate that TRP19 and TRP36 serology can provide valuable information regarding E. canis exposure and the potential genotype(s) involved in infection.

Ticks and associated pathogens from dogs in northern Vietnam.

The medical and veterinary significance of ticks and tick-borne pathogens (TBPs) in tropical and subtropical zones is well recognized. Although ticks and TBPs are known to occur in Southeast Asia, limited data is available in the international literature for some countries, such as Vietnam. The aim of this study was to investigate the species of ticks and TBPs associated with dogs in northern Vietnam. Out of 359 dogs enrolled in this study, 26.2% (n = 94) were infested by 466 ticks (i.e., 287 males, 139 females, 30 nymphs, and 10 larvae). All ticks were morphologically identified as Rhipicephalus sanguineus sensu lato, and some of them genetically characterized as belonging to the tropical lineage. A total of 302 ticks were molecularly screened for the detection of selected TBPs. Three ticks were positive for Hepatozoon canis, one for Ehrlichia canis, and one for Babesia vogeli, representing the first molecular characterization of these pathogens in Vietnam. In conclusion, the tropical lineage of R. sanguineus s.l. is the dominant tick taxon infesting dogs from northern Vietnam, where different TBPs are circulating.

Molecular Evidence for Transstadial Transmission of Ehrlichia canis by Rhipicephalus sanguineus sensu lato Under Field Conditions.

This study investigated possible transstadial transmission of Ehrlichia canis by Rhipicephalus sanguineus sensu lato collected from shelter dogs and the shelter grounds in Diyarbakir Province of south-eastern Turkey. Totally 225 engorged nymphs were collected from eight infected dogs with E. canis and incubated at 28°C for moulting. Unfed ticks from the shelter grounds comprising 1,800 larvae, 3,100 nymphs, and 85 adults were sorted according to sampling origin, life stage, and sex into 116 pools and screened by 16S rRNA PCR. Nine out of 26 pools of unfed adult ticks were positive for E. canis, with overall infection rate maximum likelihood estimation (MLE) of 4.83 (CI 2.39-8.87). E. canis was detected in three of 12 male pools (MLE 3.22, CI 0.86-8.83) and six of 14 female pools (MLE 6.16, CI 2.59-12.90). No adult pools collected from the shelter grounds were positive. Among 62 unfed nymph pools collected from the shelter, six were infected with E. canis (MLE 0.20, CI 0.08-0.42). No E. canis DNA was detected in any of the larva pools. Our results revealed molecular evidence for transstadial transmission of E. canis by R. sanguineus s.l. both from larva to nymph and from nymph to adult. We found no evidence of transovarial transmission.

Occurrence of Leishmania infantum in the central nervous system of naturally infected dogs: Parasite load, viability, co-infections and histological alterations.

Zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and little is known about the occurrence and pathogenesis of this parasite in the CNS. The aims of this study were to evaluate the occurrence, viability and load of L. infantum in the CNS, and to identify the neurological histological alterations associated with this protozoan and its co-infections in naturally infected dogs. Forty-eight Leishmania-seropositive dogs from which L. infantum was isolated after necropsy were examined. Cerebrospinal fluid (CSF) samples were analyzed by parasitological culture, quantitative real-time PCR (qPCR) and the rapid immunochromatographic Dual Path Platform test. Brain, spinal cord and spleen samples were submitted to parasitological culture, qPCR, and histological techniques. Additionally, anti-Toxoplasma gondii and anti-Ehrlichia canis antibodies in serum and distemper virus antigens in CSF were investigated. None of the dogs showed neurological signs. All dogs tested positive for L. infantum in the CNS. Viable forms of L. infantum were isolated from CSF, brain and spinal cord in 25% of the dogs. Anti-L. infantum antibodies were detected in CSF in 61% of 36 dogs. Inflammatory histological alterations were observed in the CNS of 31% of the animals; of these, 66% were seropositive for E. canis and/or T. gondii. Amastigote forms were associated with granulomatous non-suppurative encephalomyelitis in a dog without evidence of co-infections. The highest frequency of L. infantum DNA was observed in the brain (98%), followed by the spinal cord (96%), spleen (95%), and CSF (50%). The highest L. infantum load in CNS was found in the spinal cord. These results demonstrate that L. infantum can cross the blood-brain barrier, spread through CSF, and cause active infection in the entire CNS of dogs. Additionally, L. infantum can cause inflammation in the CNS that can lead to neurological signs with progression of the disease.

Ferritin 1 silencing effect in Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) during experimental infection with Ehrlichia canis.

Rhipicephalus sanguineus sensu lato (s.l.) is a very common ectoparasite of domestic dogs able to transmit several pathogens of human and veterinary importance. Tick infestations and tick-borne diseases (TBDs) remain a serious and persistent problem, due to the lack of efficient control measures. It is therefore vital that novel approaches to control are pursued. Whilst vaccination is recognised as a potential control method to reduce tick infestation, no anti-R. sanguineus vaccine is available. Ticks depend on their blood meals to obtain nutrients and to achieve sexual maturity, which exposes them to vast amounts of iron. Although an essential molecule for several biological processes, its excess can lead to oxidative stress. Iron homeostasis is achieved with the help of iron-binding proteins called ferritins, among others, present in several tick tissues and developmental stages. These evolutionarily conserved proteins regulate iron homeostasis by storing and releasing iron in a controlled manner. In this study the R. sanguineus ferritin 1 gene was silenced through RNA interference (RNAi) in adult females exposed to an experimental infection with Ehrlichia canis. The purpose of this study was to assess the role of this protein in tick feeding, ovary development, oogenesis, and pathogen acquisition. Our data has shown that silencing ferritin 1 alters tick competence to normally engorge and causes morphologic and histochemical changes in the ovaries (OV) and oocytes. Furthermore, our data revealed that no E. canis DNA was found in either experimental group. Determining the function of molecules that act in key biological processes, such as blood digestion or reproduction, and that could be considered potential tick antigens will contribute towards the improvement of current control measures against these ectoparasites and the pathogens they vector.

Cardiac troponin I concentrations, electrocardiographic and echocardiographic variables remained unchanged in dogs experimentally infected with Ehrlichia canis.

Canine monocytic ehrlichiosis (CME, Ehrlichia canis) has occasionally been associated with myocardial injury. The aim of the present study was to serially measure and evaluate cardiac troponin I (cTnI) concentrations in dogs with experimentally induced acute and subclinical CME and to evaluate potential associations between cTnI concentration and an array of echocardiographic and electrocardiographic parameters. Serum cTnI concentration and simultaneous echocardiographic and electrocardiographic recordings were evaluated in 12 healthy Beagle dogs prior to experimental infection and on days 20 and 90 post-inoculation with E. canis. Almost all serum cTnI concentrations were below the limit of detection and selected electrocardiographic and echocardiographic parameters remained unchanged throughout the study.

Comparative efficacy of oral administrated afoxolaner (NexGard™) and fluralaner (Bravecto™) with topically applied permethrin/imidacloprid (Advantix(®)) against transmission of Ehrlichia canis by infected Rhipicephalus sanguineus ticks to dogs.

BACKGROUND: The ability of the topical spot-on Advantix(®) (50 % permethrin/10 % imidacloprid) to prevent transmission of Ehrlichia canis by infected Rhipicephalus sanguineus ticks to dogs has previously been reported. The recent market introduction of chewable tablets containing the novel compounds, afoxolaner (NexGard™) and fluralaner (Bravecto™) enabled us to conduct a comparative efficacy study with respect to the ability of these three products to block transmission of E. canis by ticks to dogs. The speed of kill, immediate drop-off rate and anti-attachment efficacy of the respective products were also studied. METHODS: The study was a blinded parallel group design, wherein 32 dogs were randomised into four different groups of eight dogs. Group 1 served as negative placebo control, group 2 and 3 were treated on Days 0, 28 and 56 with NexGard™ and Advantix(®), respectively. Group 4 was dosed once on Day 0 with Bravecto™. For tick efficacy assessments 50 non-infected ticks were placed onto the dogs on Days 30, 35, 42, 49, 56, 63, 70, 77 and 84 and on animal tick counts were performed at 3 h, 6 h and 12 h after infestation. To evaluate the ability to block transmission of E. canis, each dog was challenged by releasing 80 adult E. canis-infected R. sanguineus ticks into their sleeping kennels on Days 31, 38, 45 and 52. The animals were monitored for clinical signs of monocytic ehrlichiosis (pyrexia and thrombocytopenia) and were tested for E. canis DNA by PCR and for specific antibodies using IFA. A dog was considered infected with E. canis if both PCR and IFA yielded positive test results up to Day 84. RESULTS: Mean arithmetic tick counts on dogs treated with the Advantix(®) spot-on were significantly (P < 0.0005) lower throughout the study as compared with the negative controls and was, with respect to the speed of kill and resulting onset of acaricidal efficacy, superior over NexGard™ and Bravecto™ at all time points in the 12 h period observed (3 h, 6 h and 12 h). None of the dogs treated with the Advantix(®) spot-on became infected with E. canis, whereas six out of eight untreated control dogs acquired the infection. Furthermore, E. canis infection was diagnosed in four out of eight dogs treated with NexGard™ and in two out of eight dogs treated with Bravecto™. CONCLUSIONS: The speed of kill of the two recently registered systemic compounds against R. sanguineus was not sufficiently fast to prevent transmission of E. canis and resulted in only low partial blocking and protection capacity while Advantix(®) effectively blocked transmission of E. canis to dogs in the challenge period and thus provided adequate protection for dogs against monocytic ehrlichiosis.

Comparative Evaluation of the Vector Competence of Four South American Populations of the Rhipicephalus sanguineus Group for the Bacterium Ehrlichia canis, the Agent of Canine Monocytic Ehrlichiosis.

This study compared the vector competence of four populations of Rhipicephalus sanguineus group ticks for the bacterium Ehrlichia canis, the agent of canine monocytic ehrlichiosis (CME). Ticks (larvae and nymphs) from the four populations-one from São Paulo state, southeastern Brazil (BSP), one from Rio Grande do Sul state, southern Brazil (BRS), one from Argentina (ARG), and one from Uruguay (URU)-were exposed to E. canis infection by feeding on dogs that were experimentally infected with E. canis. Engorged ticks (larvae and nymphs) were allowed to molt to nymphs and adults, respectively, which were tested by molecular analysis (E. canis-specific PCR assay) and used to infest naïve dogs. Through infestation of adult ticks on naïve dogs, after nymphal acquisition feeding on E. canis-infected dogs, only the BSP population was shown to be competent vectors of E. canis, i.e., only the dogs infested with BSP adult ticks developed clinical illness, seroconverted to E. canis, and yielded E. canis DNA by PCR. This result, demonstrated by two independent replications, is congruent with epidemiological data, since BSP ticks were derived from São Paulo state, Brazil, where CME is highly endemic. On the other hand, BRS, ARG, and URU ticks were derived from a geographical region (South America southern cone) where CME has never been properly documented. Molecular analysis of unfed adults at 30 days post molting support these transmission results, since none of the BRS, ARG, and URU ticks were PCR positive, whereas 1% of the BSP nymphs and 31.8% of the BSP adults contained E. canis DNA. We conclude that the absence or scarcity of cases of CME due to E. canis in the South America southern cone is a result of vector incompetence of the R. sanguineus group ticks that prevail on dogs in this part of South America.

Serum DHEA-S increases in dogs naturally infected with Ehrlichia canis.

Adrenocortical disturbances are expected in canine ehrlichiosis due to the immunological challenges caused by infection and consequent inflammation. Thus, this study aimed to evaluate the occurrence of adrenocortical hormonal alterations in dogs naturally infected with Ehrlichia canis (n = 21) as positively confirmed by the presence of anti-E. canis antibodies (Dot-ELISA) and nested PCR (nPCR). Serum dehydroepiandrosterone sulfate (DHEA-S) concentrations were assessed via ELISA before and one hour after ACTH stimulation. Another 10 healthy dogs were subjected to the same stimulation protocol and used as controls. The results revealed that baseline and post-ACTH DHEA-S concentrations were significantly greater in sick dogs, regardless of gender, and this finding illustrates the stress induced by naturally acquired ehrlichiosis in dogs.

Canine vector-borne co-infections: Ehrlichia canis and Hepatozoon canis in the same host monocytes.

The protozoon Hepatozoon canis and the rickettsia Ehrlichia canis are tick-borne pathogens, transmitted by Rhipicephalus sanguineus, which cause canine hepatozoonosis and canine monocytic ehrlichiosis, respectively. Co-infection of the same host monocytes with H. canis and E. canis confirmed by molecular characterization of the infecting agents and quantitative assessment of co-infected cells is described for the first time in three naturally-infected dogs. Blood smear evaluation indicated that at least 50% of the leukocytes infected with H. canis gamonts contained E. canis morulae. Co-infection of the same host cell demonstrated in this report suggests that infection with one pathogen may permit or enhance invasion or prolonged cellular survival of the other.

Transmission of Ehrlichia canis by Rhipicephalus sanguineus ticks feeding on dogs and on artificial membranes.

A South African strain of Ehrlichia canis was isolated and used to infect a laboratory-bred Beagle dog. Rhipicephalus sanguineus nymphs, which fed on this dog, moulted to adult ticks which carried infection rates of E. canis between 12% and 19% and were used in a series of in vivo and in vitro experiments. Five groups of 6 dogs were challenged with the infected R. sanguineus ticks, which were removed 24h, 12h, 6h or 3h after the ticks had been released onto the dogs. The animals were monitored for fever and thrombocytopenia and were considered infected if they became serologically positive for E. canis antibodies as well as PCR positive for E. canis DNA. Seven dogs became infected with E. canis in the following groups: Group 1 (24h tick challenge) 1 out of 6; Group 2 (12h) 1 of 6; Group 3 (6h) 2 of 6; Group 4 (6h) 2 of 6 and Group 5 (3h) 1 out of 6. Six of those 7 infected dogs developed fever and a significant thrombocytopenia. One dog did not show any symptoms, but seroconverted and was found PCR positive on several occasions. Five additional dogs were PCR positive on one test sample only but were not considered infected because they did not develop any specific E. canis antibodies. In vitro, R. sanguineus ticks attached and fed on bovine blood through silicone membranes with attachment rates up to 72.5% after 24h increasing to 84.2% at 72 h. The ticks transmitted E. canis as soon as 8h post application as demonstrated by E. canis DNA found in the nutritive blood medium. In conclusion, transmission of E. canis by R. sanguineus ticks starts within a few hours after attachment, which is earlier than previously thought. These findings underpin the need for acaricides to provide either a repellent, an anti-attachment and/or a rapid killing effect against ticks in order to decrease the risk of transmission of E. canis.

Influence of experimental canine ehrlichiosis on the E-ADA activity and purine levels in serum and possible functional correlations with pathogenesis.

The aim of this study was to evaluate adenosine deaminase activity and purines levels in serum of dogs experimentally infected by Ehrlichia canis. Banked serum samples of dogs divided into two groups with five animals each: healthy animals and animals infected by E. canis. The concentration of purines (adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), adenosine, inosine, hypoxanthine, xanthine and uric acid), and adenosine deaminase (E-ADA) activity in sera were evaluated. Samples were collected on days 12 and 30 post-infection (PI). The E-ADA activity showed a significant reduction on day 12 PI, and increased on day 30 PI in dogs infected with E. canis. On day 12, an increase in seric concentration of ATP, ADP and adenosine was verified, and different levels of hypoxanthine, xanthine and uric acid had a drastic reduction in infected compared healthy dogs (P<0.05). However, on day 30 PI, the levels of seric ADP and AMP decreased, unlike the concentration of xanthine and uric acid that increased significantly in infected dogs (P<0.05). Therefore, the activity of E-ADA and purine levels are altered in experimental canine ehrlichiosis, probably with the purpose of modulating the pathogenesis of the disease related to immune response, oxidative stress and coagulation disorders in acute phase.

Increase nitric oxide and oxidative stress in dogs experimentally infected by Ehrlichia canis: effect on the pathogenesis of the disease.

The aim of this study was to evaluate nitric oxide levels, lipid peroxidation, protein oxidation and glutathione reductase activity in serum of dogs experimentally infected by Ehrlichia canis. Banked serum samples of dogs divided into two groups were used: negative control (n=5) and infected by E. canis (n=5). The concentration of nitrite/nitrate (NOx), lipid peroxidation (TBARS), advanced oxidation protein products (AOPP), and glutathione reductase (GR) activity in sera were evaluated. Samples were collected on days 0, 3, 6, 18 and 30 post-infection (PI). NOx and TBARS levels were significantly (P<0.05) higher in the infected group at 18 and 30 days PI, as well as AOPP levels at 30 days PI when compared to samples from control group. The GR activity was significant (P<0.05) increased in serum of dogs infected by E. canis on days 18 and 30 PI. Based on the increased levels of NOx, TBARS, AOPP and GR activity we concluded that dogs experimentally infected by E. canis develop a state of redox imbalance and that these changes might be involved in the pathophysiology of the disease.

Ehrlichia canis (Jaboticabal strain) induces the expression of TNF-α in leukocytes and splenocytes of experimentally infected dogs.

Canine ehrlichiosis is caused by the bacterium Ehrlichia canis and is characterized by a systemic febrile disease of unknown pathogenesis. This study evaluated the expression of cytokines TNF-α, IL-10, IFN-γ, in splenic cells and blood leukocytes during the acute phase of ehrlichiosis and after treatment with doxycycline hyclate in dogs experimentally infected with the E. canis Jaboticabal strain. The study results showed a significant expression of TNF-α 18 days post-inoculation, reducing by approximately 70% after treatment. There was a unique peak of expression of IL-10 and IFN-γ 18 and 30 days post-inoculation, respectively. This study suggests that TNF-α plays a role in the pathogenesis of the acute phase of canine ehrlichiosis and that treatment with doxycycline hyclate reduces the systemic effects of this cytokine, possibly by reducing or eliminating parasitemia.

Ehrlichia canis (Jaboticabal strain) induces the expression of TNF-α in leukocytes and splenocytes of experimentally infected dogs / Amostra Ehrlichia canis (Jaboticabal) induz a expressão de TNF-α em leucócitos e esplenócitos de cães experimentalmente infectados

Canine ehrlichiosis is caused by the bacterium Ehrlichia canis and is characterized by a systemic febrile disease of unknown pathogenesis. This study evaluated the expression of cytokines TNF-α, IL-10, IFN-γ, in splenic cells and blood leukocytes during the acute phase of ehrlichiosis and after treatment with doxycycline hyclate in dogs experimentally infected with the E. canis Jaboticabal strain. The study results showed a significant expression of TNF-α 18 days post-inoculation, reducing by approximately 70 percent after treatment. There was a unique peak of expression of IL-10 and IFN-γ 18 and 30 days post-inoculation, respectively. This study suggests that TNF-α plays a role in the pathogenesis of the acute phase of canine ehrlichiosis and that treatment with doxycycline hyclate reduces the systemic effects of this cytokine, possibly by reducing or eliminating parasitemia.

A erliquiose canina é causada pela bactéria Ehrlichia canis, que desencadeia no hospedeiro uma doença febril e sistêmica, de patogênese pouco conhecida. O presente estudo avaliou a expressão das citocinas TNF-α, IL-10, IFN-γ, em células esplênicas e em leucócitos sanguíneos, durante a fase aguda da erliquiose e após o tratamento com hiclato de doxiciclina, em cães experimentalmente infectados com a amostra E. canis Jaboticabal. Os resultados mostraram expressão significativa de TNF-α 18 dias após a inoculação, reduzindo aproximadante 70 por cento após o tratamento. Houve um único pico de expressão de IL-10 e de IFN-γ entre 18 e 30 dias após a inoculação, respectivamente. Este estudo sugere que o TNF-α participa da patogenia da fase aguda da erliquiose canina, e que o tratamento com hiclato de doxiciclina reduz os efeitos sistêmicos dessa citocina, possivelmente por reduzir ou eliminar a parasitemia.

Efficacy of a doxycycline treatment regimen initiated during three different phases of experimental ehrlichiosis.

Doxycycline is the treatment of choice for canine monocytic ehrlichiosis (CME), a well-characterized disease and valuable model for tick-borne zoonoses. Conflicting reports of clearance of Ehrlichia canis after treatment with doxycycline suggested that the disease phase during which treatment is initiated influences outcomes of these treatments. The purpose of this study was to evaluate the efficacy of a 28-day doxycycline regimen for clearance of experimental E. canis infections from dogs treated during three phases of the disease. Ten dogs were inoculated with blood from E. canis carriers and treated with doxycycline during acute, subclinical, or chronic phases of CME. Daily rectal temperatures and semiweekly blood samples were monitored from each dog, and Rhipicephalus sanguineus ticks were acquisition fed on each dog for xenodiagnosis. Blood collected from dogs treated during acute or subclinical CME became PCR negative for E. canis as clinical parameters improved, but blood samples collected from dogs treated during chronic CME remained intermittently PCR positive. R. sanguineus ticks fed on dogs after doxycycline treatments became PCR positive for E. canis, regardless of when treatment was initiated. However, fewer ticks became PCR positive after feeding on two persistently infected dogs treated with doxycycline followed by rifampin, suggesting that antibiotic therapy can reduce tick acquisition of E. canis.

Serological study of selected vector-borne diseases in shelter dogs in central Spain using point-of-care assays.

We evaluated the prevalence of selected vector-borne diseases in 131 dogs in an animal shelter in central Spain using point-of-care assays (SNAP 4DX and SNAP Leishmania; IDEXX Laboratories, Westbrook, ME). The SNAP 4DX detects Dirofilaria immitis (Di) antigen and antibodies against Ehrlichia canis (Ec), Borrelia burgdorferi (Bb), and Anaplasma phagocytophylum (Aph); the SNAP Leishmania kit detects antibodies against Leishmania infantum (Li). Dogs were classified as healthy or sick based on physical examination, complete blood counts, and serum chemistry profiles. The prevalence of positive test results was as follows: Ec, 5.3% (n = 7); Aph, 19.0% (n = 25); Bb, 0%; Di, 0%; and Li, 5.3% (n = 7). Four dogs (3%) were coexposed to Ec and Aph, and three dogs (2.3%) were coexposed to Aph and Li. There was no statistically significant correlation between positive serology and clinical status (sick vs. healthy) or hematologic/biochemical abnormalities. The prevalence of Aph was the highest and is in agreement with a recent report in a dog shelter in northwestern Spain. These point-of-care assays may be more valuable as epidemiologic than as clinical tools.

Ticks and haemoparasites of dogs from Praia, Cape Verde.

In February 2008 an epidemiological field study on arthropod-borne infections in dogs was carried out in Praia, the capital city of Cape Verde. For this purpose 130 dogs were included in the study. Of these, 94.6% were infested with ticks. Altogether, 1293 ticks of the genus Rhipicephalus (in all evaluated cases R. sanguineus) were collected. Examination for haemotropic parasites was performed via polymerase chain reaction (PCR). Lymph node fine-needle aspirates were screened by PCR for Leishmania infantum infections in 20 dogs with enlarged lymph nodes. Our investigation revealed two species of protozoa (Babesia canis vogeli and Hepatozoon canis) and two species of rickettsiae (Anaplasma platys and Ehrlichia canis). In 101 dogs (77.7%) DNA of one or more pathogens was detected. The PCR examination for H. canis was positive in 83 dogs (63.8%), for E. canis in 34 dogs (26.2%), for A. platys in 10 dogs (7.7%) and for B. canis in five dogs (3.8%), whereas neither B. gibsoni nor L. infantum DNA could be detected. Of the infected dogs, 71.3% had a monoinfection, 27.7% had infections with two pathogens and 1.0% with three pathogens. B. canis, H. canis, E. canis, A. platys and their vector tick R. sanguineus are endemic to Cape Verde and can be present in dogs in high prevalences. These results outline the risk of importing tropical canine diseases when Capeverdian stray dogs are taken to Europe.

Host surveys, ixodid tick biology and transmission scenarios as related to the tick-borne pathogen, Ehrlichia canis.

The ehrlichioses have been subject to increasing interest from veterinary and public health perspectives, but experimental studies of these diseases and their etiologic agents can be challenging. Ehrlichia canis, the primary etiologic agent of canine monocytic ehrlichiosis, is relatively well characterized and offers unique advantages and opportunities to study interactions between a monocytotropic pathogen and both its vertebrate and invertebrate hosts. Historically, advances in tick-borne disease control strategies have typically followed explication of tick-pathogen-vertebrate interactions, thus it is reasonable to expect novel, more sustainable approaches to control of these diseases as the transmission of their associated infections are investigated at the molecular through ecological levels. Better understanding of the interactions between E. canis and its canine and tick hosts would also elucidate similar interactions for other Ehrlichia species as well as the potential roles of canine sentinels, reservoirs and models of tick-borne zoonoses. This article summarizes natural exposure studies and experimental investigations of E. canis in the context of what is understood about biological vectors of tick-borne Anaplasmataceae.

Surveillance for zoonotic vector-borne infections using sick dogs from southeastern Brazil.

For many vector-borne organisms, dogs can be used as sentinels to estimate the risk of human infection. The objective of this study was to use dogs as sentinels for multiple vector-borne organisms in order to evaluate the potential for human infection with these agents in southeastern Brazil. Blood from 198 sick dogs with clinicopathological abnormalities consistent with tick-borne infections were selected at the São Paulo State University Veterinary Teaching Hospital in Botucatu and tested for DNA and/or antibodies against specific vector-borne pathogens. At least one organism was detected in 88% of the dogs, and Ehrlichia canis DNA was amplified from 78% of the blood samples. Bartonella spp. seroreactivity was found in 3.6%. Leishmania chagasi antibodies were detected in 1% of the dogs. There was no serological or polymerase chain reaction evidence of infection with Anaplasma phagocytophilum, Borrelia burgdorferi, Ehrlichia chaffeensis, Ehrlichia ewingii, and Rickettsia rickettsii. The full E. canis 16S rRNA gene sequence of one of the Brazilian strains obtained in this study was identical to the causative agent of human ehrlichiosis in Venezuela. Ehrlichia canis may pose a human health hazard and may be undiagnosed in southeastern Brazil, whereas exposure to the other organisms examined in this study is presumably infrequent.