Serum acute phase proteins as clinical phase indicators and outcome predictors in naturally occurring canine monocytic ehrlichiosis

Background: Canine monocytic ehrlichiosis (CME), caused by Ehrlichia canis, is an important tick‐borne disease of global importance. Currently, limited information is available on the diagnostic and prognostic value of acute phase proteins (APPs) in dogs naturally infected with E. canis. Hypothesis: APPs may be useful indicators of the clinical phase of CME and predictive of the clinical outcome (death or survival). Animals: Fifty‐six dogs naturally infected with E. canis and 7 clinically healthy control dogs. Methods: C‐reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), and albumin concentrations determined on admission were retrospectively compared among 27 dogs with nonmyelosuppressive CME, 29 dogs with myelosuppressive CME and 7 healthy dogs. Diagnosis of CME was based on clinical and clinicopathological findings, seropositivity to E. canis, polymerase chain reaction amplification of E. canis‐specific 16S rDNA, microscopic observation of Ehrlichia sp. morulae in blood monocytes or some combination of these. Results: Mean concentrations of CRP, SAA, and Hp were significantly higher in the myelosuppressed dogs compared with the other groups, but no significant differences were found in the concentration of albumin. Survival analysis of the affected animals indicated that APP concentrations were not associated with clinical outcome; the latter was strongly associated with pancytopenia (odds ratio for death 22.7) and neutropenia (odds ratio for death 7.7). Conclusions and Clinical Importance: CRP, SAA, and Hp serum concentrations on admission are useful indicators of the clinical phase of CME, but are not useful predictors of clinical outcome.     

Outbreak of canine monocytic ehrlichiosis in Saudi Arabia

BACKGROUND: Canine monocytic ehrlichiosis (CME) is a widespread tickborne infection of canids caused by Ehrlichia canis, a gram-negative obligatory intracellular bacteria belonging to the family Anaplasmataceae. CME is reported to have worldwide distribution, but its presence in a region requires the presence of the vector, the brown dog tick Rhipicephalus sanguineus. OBJECTIVE: This purpose of this report was to describe an outbreak of CME in a colony of dogs resident in the eastern region of Saudi Arabia. METHODS: History, presenting clinical signs, and the results of a CBC, biochemical panel, and serology (using a commercial test for E canis antibodies) were evaluated in 9 male Labrador Retrievers between October and December 2006. RESULTS: The majority of dogs presented with severe lethargy (7/9) and acute anorexia (5/9), and had fever (7/9) and generalized lymphadenopathy (7/9). The most common laboratory abnormalities were anemia (8/9), leukopenia (7/9), and hypoalbuminemia (6/9). Thrombocytopenia was found in only 2 dogs, 1 of which had increased bleeding tendency. Ehrlichia morulae were identified in blood films from 4/9 dogs and serologic test results were positive in 7/9 dogs. Confirmation of Ehrlichia sp infection was obtained in 1 dog by using a genus-specific real-time PCR assay. Four dogs had tick infestation; the ticks on 1 dog were identified as R sanguineus. All of the dogs had a rapid clinical response to doxycycline hyclate. CONCLUSIONS: This report, to our knowledge, is the first to describe the presence of a pathogenic Ehrlichia organism affecting dogs in Saudi Arabia. Additional molecular studies are necessary to confirm E canis infection, and to identify the strain of the organism.     

Cytologic patterns of lymphadenopathy in canine monocytic ehrlichiosis

Background: Recognition of different cytologic patterns in lymph nodes (LNs) from dogs with canine monocytic ehrlichiosis (CME) and noninfectious causes of lymphoid reactivity may have diagnostic utility. Objectives: The aims of the present study were to compare cytologic patterns in LNs of dogs with different phases of CME, to investigate the association of cytologic pattern and presence of Ehrlichia spp. morulae, and to compare patterns of lymphoid reactivity between dogs with CME and those with noninfectious causes of lymphoid hyperplasia. Methods: Cytologic preparations of LNs from 35 dogs with nonmyelosuppressive CME (group A), 16 dogs with myelosuppressive CME (group B), 26 dogs with noninfectious diseases (group C), and 15 healthy dogs (group D) were evaluated. Percentages of lymphocyte types, plasma cells, macrophages, neutrophils, and eosinophils were determined. Samples from dogs in groups A and B were evaluated for the presence of morulae. Results: Cytologic abnormalities in LNs were recorded in 54% of dogs in group A, 88% in group B, 39% in group C, and 0% in group D and were more frequent (P=.02) in dogs with myelosuppressive CME than those with nonmyelosuppressive CME. Plasma cell hyperplasia was more frequent in CME than in noninfectious diseases (P=.03). An association between the presence of cytologic abnormalities and morulae in group A dogs was not found. Conclusions: Dogs with myelosuppressive CME have more lymphoid cytologic abnormalities than dogs with nonmyelosuppressive CME. LN plasmacytosis is the major pattern of lymphadenopathy in dogs with CME and is found more frequently in dogs with CME than in dogs with noninfectious causes of lymphadenopathy.     

Evaluation of neutrophil oxidative metabolism in canine monocytic ehrlichiosis

BACKGROUND: Canine monocytic ehrlichiosis (CME) is a tick-borne disease caused by Ehrlichia canis, a rickettsia that infects the monocytes of dogs. This infection can result in a chronic and life-threatening disease. Thrombocytopenia, mild anemia, and leukopenia are the most common hematologic findings in CME. OBJECTIVE: To investigate the role of peripheral blood neutrophils in CME, an evaluation was conducted of their functional state during the acute phase of the disease in dogs experimentally infected by E canis. METHODS: Seven dogs were inoculated with E canis, and 3 remained as uninfected controls. All dogs had physical exams and hematologic tests (CBC and nitroblue tetrazolium [NBT] reduction) during a 6-week period. RESULTS: There was no difference (P > .05) in spontaneous NBT reduction results between the 2 groups of dogs throughout the 6-week period of observation. Nevertheless, when stimulated, the neutrophils showed higher activity in the infected group (P = .01) on weeks 4 and 5 after infection. CONCLUSION: Infection by E canis has no influence on neutrophil oxidative metabolism even though during the remission period of the acute phase of the disease, the neutrophils seem to be more reactive under stimulation.     

Seroepidemiology of canine babesiosis and ehrlichiosis in a hospital population

Canine ehrlichiosis and babesiosis have a worldwide distribution with geographic variation in prevalence and main clinical manifestations. We prospectively determined seroprevalence of canine babesiosis and ehrlichiosis, and risk factors for seropositivity. Three hundred and eighty-one dogs were randomly selected to represent the canine population at a Veterinary Teaching Hospital in south Brazil (latitude 23° S). Dogs were tested with a point-of-care ELISA for Ehrlichia canis antibodies and IFA to confirm previous exposure to Babesia vogeli. Multiple logistic regression analysis was then used to estimate adjusted odds ratio (OR) and their 95% confidence intervals. One hundred and thirty-six (36%) dogs were seropositive for B. vogeli antibodies, whereas 87 (23%) dogs were seropositive to E. canis antibodies. Fifty-four (14%) dogs seroreacted to both agents. Adult dogs previously infested with ticks were more likely to seroreact to B. vogeli or E. canis. Superficial bleeding (OR = 12.4) was more common in dogs exposed to B. vogeli, whereas neurological signs (OR = 7.7) were more common in dogs seropositive to E. canis. Neurological signs (OR = 12.0) and lameness (OR = 12.8) were more prevalent in dogs that seroreacted to both organisms. Owners of dogs with ticks were more likely to have been exposed to ticks themselves (OR = 3.2). Canine babesiosis and ehrlichiosis appear to be highly prevalent in this hospital population. Clinical signs differed from the most common signs in other regions with bleeding occurring more in dogs seropositive to babesiosis, but not ehrlichiosis; neurologic signs in dogs with E. canis antibodies; and lameness in dogs that seroreacted to both organisms.     

Platelet aggregation responses in clinically healthy adult llamas

Background: Limited information exists regarding hemostasis in camelids despite the importance of platelet function testing in the accurate identification of platelet disorders. As further importation of llamas to North America is restricted, variability in breeding stock will continue to decrease, potentially leading to an increase in heritable bleeding disorders. Objective: The objective of this study was to measure platelet aggregation responses in clinically healthy llamas and provide baseline data to which abnormal platelet function may be compared in the future. Methods: Blood samples were collected from 39 healthy adult llamas, citrated, and centrifuged to produce platelet‐rich plasma (PRP). Within 4 hours of the blood draw, 20 μL of each agonist reagent were added to 180 μL of PRP. Final concentrations of agonists were 2 × 10^?5 M ADP, 0.19 mg collagen/mL PRP, 1 × 10^?4 M epinephrine, and 500 μg arachidonic acid/mL PRP. Results: Llama platelets were most responsive to ADP and collagen, with a maximum percent aggregation (mean±SD) of 71.3±18.6% and 55.8±19% and aggregation rates of 9.5±3.9 and 6.7±3.7 cm/min, respectively. Llama platelet aggregation in response to epinephrine and arachidonic acid was minimal to absent. Conclusions: This study is the first of its kind to establish baseline values for platelet aggregation in healthy adult llamas.     

广州地区犬场工作人员与一般人群犬埃立克体感染的比较研究

从比格犬埃立克体阳性血清中提取犬埃立克体IgG,用此IgG免疫鸡制备出抗抗体,用已建立的ELISA检测法对采集的人血清样本进行检测。143份随机采集的人血清4份呈阳性,阳性率2.78%;52份犬场工作人员血清6份呈阳性,阳性率11.54%。结果表明,广州地区犬场工作人员较一般人群犬埃立克体感染率高。     

Ocular manifestations of natural canine monocytic ehrlichiosis (Ehrlichia canis): a retrospective study of 90 cases

OBJECTIVE: To investigate the spectrum, prevalence and treatment response rate of ocular manifestations associated with natural canine monocytic ehrlichiosis (CME) due to Ehrlichia canis. METHODS: The medical records of 90 dogs that were admitted for primary ocular problems and reacted positive for E. canis serum antibodies were reviewed. All the dogs were treated with oral doxycycline. In 79 dogs, a combination of systemic corticosteroids and/or topical anti-inflammatory, mydriatics/cycloplegics and antibacterial agents were applied. RESULTS: The dogs ranged from 0.5 to 15 years in age; 55 were male and 35 female. A total of 19 breeds were represented. In the majority of dogs, ocular abnormalities were noticed in conjunction with other CME-induced clinical signs. In 30/90 dogs, ophthalmic signs were the sole presenting complaint. Unilateral (22/90, 24.5%) and bilateral (68/90, 75.5%) uveitis was the most common ophthalmic diagnosis and was classified as anterior, posterior or panuveitis in 58 (64.5%), 8 (8.9%) and 24 (26.6%) of the dogs, respectively. Bilateral uveitis was significantly more common than unilateral uveitis (P < 0.0001), and anterior significantly more common than posterior uveitis (P < 0.0001) or panuveitis (P < 0.0001). In addition, corneal ulceration (12/90, 13.3%), necrotic scleritis (10/90, 11.1%), low tear production (8/90, 8.9%) and orbital cellulitis (3/90, 3.3%) were seen. Of the 45 (50%) dogs with a satisfactory follow-up, 25 (55.5%), 11 (24.5%), and 9 (20%) showed complete, partial and poor response to treatment, respectively. The number of dogs exhibiting complete resolution of the ocular manifestations was significantly higher than those with partial (P < 0.0001) or poor (P < 0.0001) response. CONCLUSIONS: CME should be considered a major differential for a wide range of ocular manifestations exhibited by dogs residing in the endemic areas of the disease. Anterior bilateral uveitis appears to be the most prevalent ocular lesion and a favorable outcome to systemic and topical treatment may be expected in the majority of the affected dogs.     

Evaluation of a serum-based PCR assay for the diagnosis of canine monocytic ehrlichiosis

The aim of this study was to estimate the relative diagnostic sensitivity and specificity of a polymerase chain reaction (PCR) assay in the serum of dogs with naturally occurring non-myelosuppressive canine monocytic ehrlichiosis (CME), and to investigate the association between PCR positivity and immunofluorescence antibody (IFA) titres for Ehrlichia canis. Serum samples obtained from 38 dogs with non-myelosuppressive CME and 12 healthy dogs were analyzed retrospectively. Each serum sample was analyzed in triplicate using an E. canis-specific nested PCR assay targeting a 389 bp sequence of the 16S rRNA gene. E. canis DNA was amplified in 24 of 38 (63.1%) affected dogs; all samples from healthy dogs were negative. A high level of agreement was found among the PCR replicates (P < 0.0001). Median IFA titre of the 24 PCR-positive dogs was significantly lower than that of the PCR-negative infected dogs (P = 0.0029), indicating that E. canis DNA may circulate prior to the development of a high antibody titre. Serum-based PCR analysis is suggested for the early diagnosis of CME when whole blood samples are not available.     

Platelet aggregation in dogs experimentally infected with Rickettsia rickettsii

Platelet aggregation studies were performed on nine Beagle dogs experimentally infected with Rickettsia rickettsia. Platelets from dogs with Rocky Mountain spotted fever tended to be more aggregable than controls.     

Bone marrow mastocytosis in dogs with myelosuppressive monocytic ehrlichiosis (Ehrlichia canis): a retrospective study

BACKGROUND: Bone marrow mastocytosis has been reported rarely in naturally occurring canine monocytic ehrlichiosis (CME). OBJECTIVES: The aims of the present study were to estimate the prevalence and magnitude of bone marrow mastocytosis in a case series of dogs with natural CME and to assess the association, if any, between mastocytosis and the clinical severity of the disease. METHODS: Seventy-six dogs with confirmed CME (Ehrlichia canis) were included in the study. Affected dogs were allocated into group A (n = 51) without bone marrow hypoplasia and group B (n = 25) with bone marrow hypoplasia. Twenty clinically healthy Beagles not previously exposed to E canis served as controls (group C). The main inclusion criteria for group A were documentation of normocellular to hypercellular bone marrow and complete clinical cure following a 4-week treatment with doxycycline, while those for group B were bone marrow hypoplasia and lack of response to doxycycline. Bone marrow aspirate smears from all 96 dogs were Giemsa-stained and examined for the presence of mast cells, which were calculated as a percentage of 1,000 nucleated cells (NCs). The prevalence of mastocytosis was compared among the 3 groups by the Pearson's chi-square test. RESULTS: Bone marrow mastocytosis (>0.1% of NCs) was found in 5 (20%) dogs in group B (range, 0.5-2.5% of NCs; median, 1% of NCs). One dog in each of groups A and C had 0.1% mast cells in the marrow. The prevalence of bone marrow mastocytosis in dogs in group B was significantly higher (P = .004) than in groups A and C. CONCLUSION: Bone marrow mastocytosis can be seen in a substantial number of dogs with E canis-induced myelosuppression.     

Usefulness of a rapid immuno-migration test for the detection of canine monocytic ehrlichiosis in Africa

A rapid immuno-migration test for the serological detection of canine monocytic ehrlichiosis, Witness^® Ehrlichia (WE) (Zoetis, France), was evaluated in 528 serum samples from dogs living in endemic areas of West and East Africa: Senegal (N = 208), Ivory Coast (N = 7), Sudan (N = 27), and Djibouti (N = 286). Of these dogs, 200 were French military working dogs (MWD) temporarily residing in Africa. The WE test results were compared with those obtained by indirect immunofluorescence (IFA). The sensitivity of WE was 97% [94.2, 98.7] with a specificity of 100% [98.6, 100]. In MWD, the seroprevalence (IFA) was 7%; in native dogs, it reached 77.1%. This significant difference can be explained by prophylactic measures from which MWD benefit. The WE test represents a simple, fast and reliable test for the detection of anti-Ehrlichia canis antibodies. Its implementation for the diagnosis of clinical cases has been validated in the field, and its use allows easy detection of asymptomatic dogs that may be carriers of E. canis.     

Prospective comparative study of 3 treatment protocols using doxycycline or imidocarb dipropionate in dogs with naturally occurring ehrlichiosis

This paper reports the clinicopathologic responses of 93 dogs with spontaneously occurring ehrlichiosis to 3 different treatment protocols. Thirty-two dogs were treated with doxycycline (10 mg/kg/day for 28 days), 31 were treated with imidocarb dipropionate (5 mg/kg given 15 days apart in 2 separate injections), and 30 were treated with both drugs simultaneously, at the doses as specified. The dogs underwent clinicopathologic evaluation before and after treatment, and were examined periodically during the 24-month period after the treatment. No differences were found in the clinical responses among the dogs in the 3 treatment groups. As for the clinicopathologic response, in spite of the fact that at the end of the study the results obtained with the 3 protocols were similar, the platelet count and serum protein electrophoresis results returned to normal more slowly in dogs that received imidocarb dipropionate as compared to those given the other 2 treatments.     

Sero-prevalence and risk indicators for canine ehrlichiosis in three rural areas of Brazil

Ehrlichia canis has a worldwide geographic distribution, occurring particularly in tropical and subtropical areas. In Brazil, the main vector in urban areas is believed to be the brown dog tick Rhipicephalus sanguineus, but little is known about the occurrence, transmission and other epidemiological aspects of canine ehrlichiosis in rural areas, where Amblyomma ticks are found more frequently than R. sanguineus. A sero-prevalence study of canine ehrlichiosis was carried out in three distinct rural regions of the State of Minas Gerais, Brazil. Serum samples were collected from 226 dogs living on farms in Lavras (n=85), Belo Horizonte (n=45), and Nanuque (n=96) and were analyzed by an indirect fluorescent antibody test for the detection of anti-Ehrlichia canis antibodies. Age, breed, sex, presence of ticks and packed cell volume were also recorded. There were 65.6% positive dogs in Nanuque, 37.8% in Belo Horizonte, and 24.7% in Lavras. Animals living in Nanuque were 4.6 times more likely to be serologically positive than dogs living in the other two regions and antibody titres were considerable higher in this area. Male dogs, dogs >5 years of age, those infested with ticks, and mongrels all showed higher rates of positivity. The results point to the importance of canine ehrlichiosis in rural areas and indicate the need for further studies on natural transmission and maintenance of the disease.     

我国犬埃立克体病研究现状

犬埃立克体病是由严格细胞内寄生的埃立克体所致的疾病。目前,我国已经发现的犬埃立克体病原有犬埃立克体(Ehrlichia)和扁平埃立克体(E.platy)两种。我国已发现的犬埃立克体病均为E.canis和E.platy的混合感染。临床症状包括发热、体重严重减轻、流鼻血和腹泻带血等;白细胞和血小板计数同时下降是我国犬埃立克体病的特点;严重的病犬常因广泛性出血或继发感染而死亡。犬埃立克体病的病原学诊断主要依靠镜检、血清学试验和PCR等。犬血清埃立克体抗体的调查结果表明,我国犬埃立克体病的流行与蜱活动密切相关。四环素类抗生素是治疗犬埃立克体病的有效药物。     

Seroprevalence of Ehrlichia canis infections in police dogs in Spain

A seroepidemiological study of Ehrlichia canis was made in police dogs in Madrid (Spain). AntiEhrlichia canis antibodies were detected by indirect fluorescent antibody test for the detection of IgG.

The results obtained (three positive dogs out of a population of 131 animals) represents a seroprevalence of canine ehrlichiosis of 2.29%. This seroprevalence is one of the lowest described for this type of population.

The seroprevalence obtained from hunting dogs kennelled in Madrid in 1993 was 66.7% (24 positive out of a population of 36). Environmental conditions (temperature, humidity, rainfall, etc.) in both populations were very similar. We suggest that these conflicting results are due to the different prophylactic programmes used in these two populations.     

Platelet aggregation studies in dogs with acute Ehrlichia platys infection

Ten adult dogs (5 Beagles and 5 mixed-breed dogs) were inoculated IV with canine platelets containing Ehrlichia platys. Inclusions and morulae of E platys developed in platelets of infected dogs at 10 to 14 days after inoculation, followed by marked thrombocytopenia at 14 to 21 days. Parasitemia and marked thrombocytopenia recurred at 24 to 28 days after inoculation. Increased numbers of megakaryocytes were observed in marrow aspirate smears from infected dogs, indicative of regenerative thrombocytopenia. Prior to infection, platelet-rich plasma from these dogs was determined to have similar aggregatory response to arachidonate. After infection with E platys, the aggregatory response of platelet-rich plasma to collagen or 3 dilutions of adenosine diphosphate was evaluated. A statistically significant (P less than 0.05) inhibition of platelet aggregatory response to the lowest dilution of adenosine diphosphate was detected for mixed-breed dogs, whereas aggregation responses were unchanged in Beagles. Results indicate that platelet activation may occur in dogs with acute ehrlichial infection.     

Diagnosis of canine monocytotropic ehrlichiosis (Ehrlichia canis): An overview

Canine monocytotropic ehrlichiosis (CME), caused by the rickettsia Ehrlichia canis, an important canine disease with a worldwide distribution. Diagnosis of the disease can be challenging due to its different phases and multiple clinical manifestations. CME should be suspected when a compatible history (living in or traveling to an endemic region, previous tick exposure), typical clinical signs and characteristic hematological and biochemical abnormalities are present. Traditional diagnostic techniques including hematology, cytology, serology and isolation are valuable diagnostic tools for CME, however a definitive diagnosis of E. canis infection requires molecular techniques. This article reviews the current literature covering the diagnosis of infection caused by E. canis.